The Process of Transporting Koi - Home & Events · (primarily cortisol) ... the effect of multiple stressors and the severity of continuous stressors are important factors that will

Transporting Koi

A Research Paper done for the

Koi Health Advisor Program January 09

Authors (listed alphabetically): Ken Austin, Terri Janas, Karen Pattist, Kay Rice

© 2009 Ken Austin, Terri Janas, Karen Pattist, Kay Rice

Disclaimers: The authors did not have unlimited time and money. One product was tested and it

worked. Therefore, there was only data to back up the recommendation for that one product, ULTIMATE®. This does not in any way mean that the group endorses ULTIMATE® as a

preference over other identical products. It just means that those other products were not tested and therefore there was no data to compare. A further discussion of ULTIMATE® may be found

in the Appendix.

The AKCA Board of Directors has requested that this disclaimer be added to apprised readers that the entire Board was not given the opportunity to review and comment on this report and/or

summary and therefore neither is endorsed by the AKCA.

Table of Contents

SCOPE OF THIS PAPER INTRODUCTION 1.0 STRESS 1.1 Physiological Response to Stress 1.2 Ammonia Stress 1.3 Transport Water Composition Changes and Stress 1.4 Controlling Variations in Water Parameters 1.5 Delayed Mortality Syndrome (DMS) 1.6 Other Indications of Stress in Koi 1.7 General Adaptation Syndrome (GAS) 1.8 Parallels with Human Health 2.0 STRESS RESEARCH DONE FOR THIS PAPER 3.0 PREPARATION FOR TRANSPORT 3.1 Attention to Stressors 3.2 Fasting 3.3 Holding Tank Considerations 3.4 Holding Tank 3.5 Attention to Slime Coat 3.6 ULTIMATE® and Gills 4.0 TRANSPORT CONSIDERATIONS 4.1 Minimizing Stress While Catching Koi 4.2 Quick and Efficient Transport 4.3 Organizing to Transport Koi 4.4 Bag/Transport Tank Water 4.5 Un-bagging Koi 4.6 The Recovery and Quarantine Tank 4.7 Transport Injuries 5.0 TRANSPORT ADDITIVES 5.1 Detoxification of Ammonia 5.2 Detoxification of Chlorine and Chloramines 5.3 Slime Coat Enhancement 5.4 Treatment of Osmotic Stress 5.5 Detoxification of Heavy Metals 5.6 Disinfecting the Water 5.7 Buffering the Water 5.8 Increasing Oxygen 5.9 Sedation

Table of Contents (Cont’d) 5.0 TRANSPORT ADDITIVES (Cont’d)

Page 2, TRANSPORTING KOI, Austin, Janas, Pattist and Rice

5.10 Darkness 5.11 Additive Conclusions & Recommendations 6.0 EQUIPMENT AND SUPPLIES 6.1 Equipment and Supply List 6.2 Seine net 6.3 Pole Net 6.4 Koi Bowl 6.5 Koi Bags 6.6 Rubber bands 6.7 Ice Packs 6.8 Containers 6.9 Oxygen 6.10 Emergency Kit 6.11 An Alternative – Live Haul Tanks 7.0 SUMMARY – 50 Things You Should Know about Transporting Koi 7.1 Stress and Koi 7.2 Preparation to Transport 7.3 Netting and Bowling Koi 7.4 Shipping Bags 7.5 Shipping Containers 7.6 Un-bagging Koi 7.7 Recovery and Quarantine after Transport References Glossary Appendix Table 1 Relative Ammonia Toxicity as a Function of Temperature and pH Table 2 Solubility of oxygen in water in milligrams per liter Table 3 Bagging Survey Results Table 4 Salt Amounts to get a solution of .3% table salt in a given volume of water Table 5 Recommended Tank Loading Restrictions for Koi Shows ULTIMATE® Product Data Sheet ULTIMATE® and why it was chosen for this paper Maintaining Stable pH and Calcium in a Holding, Recovery or Quarantine Tank Koi Food Notes Water Borne Antibiotics Web sites with information about moving large Koi Web sites with information about quarantine procedures Biographies


SCOPE OF THIS PAPER This paper was created to both examine the current research about transporting Koi, and to add to that body of knowledge by doing some original research. Koi are transported for many reasons, and doing it correctly involves preparation and an understanding of the factors involved. Too often, Koi become ill after transport, and many things are blamed when the primary cause was a lack of understanding of the stressors involved at each step in the process. Every effort must be made to minimize stress each time the Koi are handled. Proper equipment and supplies are discussed and methods are recommended to assure that Koi arrive at their destination in the best possible condition. Alternatives must be weighed at each step so that the health of the Koi can be maximized. Transporting Koi also involves an understanding of the ever-changing chemistry of the shipping water within the bag. The product ULTIMATE® is referred to throughout this paper, as it was the only one tested in association with this paper. There is a discussion in the Appendix regarding why this product was chosen and why only one product name is mentioned. The paper concludes with a list of recommendations for transporting Koi. Words in italics may be found in the glossary.

INTRODUCTION Koi are moved many times in their lives. Breeders move fry to grow out ponds and for culling. The longer a Koi is kept by a breeder, the more times it is moved and handled. Eventually, the Koi is purchased by a dealer and shipped to the dealer’s facility, where it may be moved and handled many more times before it is ultimately purchased by a pond owner. The hobbyist/pond owner moves the Koi from a quarantine tank to the pond. The hobbyist may also choose to transport their Koi to shows, to a veterinary surgeon, to exhibitions or seminars, to an indoor facility for the winter and an outdoor pond for the summer, or just between ponds. Transporting Koi correctly is more complicated than many realize, but there is a safe way to do it. The purpose of the paper is to provide detailed information on the safest methods to move Koi, regardless of the purpose for moving them. Failure to transport Koi correctly will cause the Koi harm and even death. This paper will consider the effects of transportation on the physiology of the Koi, and will recommend a process of how Koi should be transported to incur the least amount of harmful stress and injury in order to maintain the health and safety of the Koi.


1.0 STRESS 1.1 Physiological Response to Stress

“Fish are easily stressed by handling and transport, and stress can result in immuno-suppression, physical injury or even death. …The stress caused by handling, grading and transporting can be considerable. …The major concerns in transporting aquatic animals are the management of handling stress, mechanical shock, heat stress and water quality [1].

Stress may be defined as a condition in which the dynamic equilibrium of animal organisms, called homeostasis, is threatened or disturbed as a result of the actions of intrinsic stimuli, commonly defined as stressors [2]. Bartelme [3] says, “A simple definition of a stressor in fish could be a stimulus that requires a physiological response by the animal in an attempt to adapt to that stimulus.” Stress may be quantified as the amount of disturbance of homeostasis. Stress should be considered cumulative - becoming greater by stages or additions. Stress is typically not a single phenomenon. Stressors can be multiple or continuous and vary in intensity. Stress can have a chemical cause or a physical cause or even just a perceived threat can trigger a stress response. Koi display a wide variation in their physiological responses to stress (called stress-response), which is clearly evident in the catecholamine hormone and plasma corticosteroid changes that occur following a stressful event [4]. Catecholamine changes (primarily adrenalin and noradrenalin) are typically an immediate response to stress and cause increases in cardiac output, blood sugar, respiration, oxygen uptake, and blood flow to the gills. Corticosteroid changes (primarily cortisol) are typically a response to chronic stress and affect the Koi’s osmotic balance and metabolism. Transportation of Koi creates a combination of stressors and the response is variable and complex:

“The length of time between discrete stressors, the effect of multiple stressors and the severity of continuous stressors are important factors that will likely influence how Koi respond [2].” “Because the integrated stress response compromises many non-specific elements, most notably the drain of metabolic energy and susceptibility to diseases, many effects of stressors are additive [5].”

A single stressor may thus reduce the capacity of Koi to tolerate subsequent or additional stressors. Take, for example, a Koi that has been exposed to sub-lethal levels of ammonia. Stoskopf [6] says,

“Un-ionized ammonia levels of 0.2 ppm can cause rapid death and levels as low as 0.02 ppm cause severe chronic problems. Even levels as low as 1 ppb can irritate gills and cause temporary damage. Ammonia burns the gills and mucous membranes of the skin, mouth and intestines.”


The response to the stressors of transportation, which include handling, crowding, and confinement, exacerbates any existing impaired gill function due to the influence of Cortisol, a hormone excreted under stress. Acute stress limits the Koi’s capacity to osmoregulate (defined as keeping the body's fluids from becoming too dilute or too concentrated), and prolonged periods of extreme stress may lead to osmotic shock and death [4]. Sink et al [9] goes as far as to suggest that: “Stress and the concurrent physiologically high concentrations of Cortisol are more highly correlated to disease susceptibility than bacterial pathogen load”. The next sections examine the effects of known stressors as they relate to transporting Koi. 1.2 Ammonia Stress Ammonia is the final product of nitrogen metabolism in fish, and most of it is eliminated through the gills into the water. Fasted fish that are metabolizing mostly body fat, which contains no nitrogen, still produce ammonia, but to a much smaller extent. Ammonia exists in two molecular forms: the NH3 molecule and the NH4

+ ionic molecule. The ratio of these 2 forms depends on the pH and water temperature. The Koi’s gill epithelial cell walls (walls of the cells that form the tissues of the gills) are comparatively impermeable to the ammonia ion, but molecular ammonia (NH3) easily penetrates, and thus is toxic. As explained in the next section, the transport water pH tends to decrease and the water should be at a cool temperature. Under these conditions the ammonia in the transport water is mostly in the NH4

+ ionic form, and as a consequence, ammonia toxicity during transport should be less of a problem. Existing ammonia passes from the water through the gills to the blood and then into the tissues. Ammonia has a high affinity for the brain and Koi subjected to ammonia will exhibit nervous symptoms [10]. Both higher pH and higher temperature increases ammonia toxicity. (see Table #1, Appendix) If the elimination rate of the ammonia is reduced for any reason, the ammonia level in the blood increases, and may eventually reach a fatal level. This condition is called ammonia autointoxication [10, 11]. It is important to remember that there are two factors involved; the metabolism of nitrogen and the elimination rate of ammonia through the gills. Koi that have been fully fasted for a week are producing less ammonia. The less ammonia there is in the bloodstream of the Koi, the less likely it is that ammonia autointoxication can occur. However, Koi that are fed a standard high protein diet are producing a large volume of ammonia and require healthy gills to eliminate it from their bloodstream. When Koi are then cooled in preparation to or during shipping, the ammonia builds in the bloodstream because the cooler temperatures have reduced respiration and therefore reduced ammonia transfer from the gills. This is why autointoxication may occur during transport. The elimination rate of ammonia is a function of the health of the gill structure and the metabolic levels of the Koi. Various extremes in water parameters, toxins and stress can cause


morphological changes to the gill structure and thus reduce its ability to eliminate ammonia. The gill structure can also be damaged by parasites, bacterial, viral and fungal diseases. The metabolic level of Koi can be reduced by cold water temperatures or the use of anesthesia. At low metabolic levels, respiration is reduced and so is the ammonia elimination rate. Understanding the danger of ammonia autointoxication can help in the formulation of a few rules to safely transport Koi. Fully fast Koi for a week prior to transport. Do not use anesthetics in transport water. Do not transport Koi in cooled water when not fully fasted. Maintain high water quality during transport by: neutralizing gill damaging agents like chlorine binding ammonia during transport. Minimize stress to Koi during the entire transport process. Do not transport Koi with existing gill damage. 1.3 Transport Water Composition Changes and Stress The hobbyist should always ensure that Koi are kept in excellent water quality. This is especially true in preparation for transport because poor water quality is a stressor, and can additionally cause physical impairment, such as damage to the gills caused by ammonia, and bleeding from the gills [11, 12]. The chemistry of the water within the bag is more complicated than simply the ammonia level, because the toxicity is linked to the pH. As the Koi breathe, more and more CO2 is trapped and builds up in the bag (which would normally de-gas into the atmosphere), or it is converted into carbonic acid. Either way, the pH decreases. Because there is much less of the toxic component of the ammonia at the ever decreasing pH in the bag, Koi can actually survive a much higher ammonia level while shipping than they could in the pond. When Koi bag water is tested after a 24 hour trip from Japan to the East Coast of the United States, the ammonia level may be very high and the pH may be very low. Koi can survive this due to the small amount of toxic ammonia at the low pH. CO2 gases-off when the bag is opened, which causes a subsequent upward pH shift and a subsequent increase in ammonia toxicity. That is why Koi should be removed from the bag promptly after it is opened. Floating the bag before opening it helps Koi adapt to any change in water temperatures. Some references suggest that pH should also be equalized between the new tank/pond and the bag water, but the damage done by the exposure to ammonia far exceeds the stress incurred by rapidly changing pH. In fact, Koi acclimate to pH changes within several minutes, and while it is a stressor, it is far less detrimental than the gill burning that occurs with exposure to ammonia. 1.4 Controlling Variations in Water Parameters Water quality parameters can differ significantly from the starting pond to the final pond or tank. Additionally, the parameters in the transport container are constantly degrading with time. Changes in water quality are one source of stress during and after transport.


Proper transportation of Koi involves being aware of the water parameters at each stage of transport and being prepared to adjust or manage the variations. Source water, bag water and destination water should all be tested before transport in order to better control the stress caused by changing water quality. The goal should be to try to attain pH differences of less than 0.2 pH points and temperature differences of less than 5 Degrees F. Where adjustments need to be made to narrow the variation, it is preferable to raise the lower parameter than to lower the higher one. A water additive that contains a buffer will help to reduce stress by controlling the pH shift during transport and is beneficial so long as the ammonia being produced by the Koi is bound (by another chemical) as soon as it appears . An ice pack that is insulated and taped to the top of the shipping box will help to keep a cooler water temperature, which is also beneficial in reducing stress. 1.5 Delayed Mortality Syndrome (DMS) Rapid changes in water parameters, including pH, temperature and hardness, can stress Koi even if the changes are within the Koi’s normal range of adaptation. Adding the stress caused by relatively small water quality changes to the stress caused by netting, confining and transporting Koi can result in death. Although the Koi may survive the transport and seem fine in the new environment, they can become sick several days to even weeks later. This is called Delayed Mortality Syndrome. It is often associated with opportunistic infections which take advantage of the stressed Koi’s diminished immune system. According to Noga [13],

“While infections that are caused by delayed mortality syndrome are usually evident within 2-5 days of the stressful event, they may not appear until over 1 week later. A critical period of about 2 to 3 weeks after the acute stress is the most likely time that fish will become sick because of DMS; thus close observation is warranted.”

The mechanism responsible for DMS is not well understood, although it is observed in most species of fish. While Koi are considered relatively resistant to DMS compared to other species, it is always prudent to quarantine Koi after shipping. During that time, Koi should be closely inspected for disease, and water may be salted to 0.3% (see Table #4, Appendix) if the Koi seem stressed or injured in order to ease the osmoregulatory load [13] so that the Koi have more energy reserves that they can recruit for healing. 1.6 Other Indications of Stress in Koi Stress can manifest itself differently in each Koi. Stress may be exhibited in Koi as general redness in the Koi’s skin, fins or bleeding from the gills. Stress causes capillaries to become visible because they are engorged with blood due to increased blood pressure. It can be seen as overall redness or red veining in the white skin of the Koi. Some Koi bleed from their gills [22] during a particularly stressful event. While neither of these symptoms will kill Koi directly, they indicate the extent of the problem. The altered chemistry and residual effects of stress are well known. One indicator of a Koi’s stress level is the level of cortisol in the blood [2, 9]. Cortisol levels in the blood increase after stressful situations, and significant mortality


rates have been seen in test subjects with increased cortisol levels [9]. A study by Svobodova et al [14] also confirms that the stress hormone cortisol is highly correlated with the increased susceptibility of fish to infection and mortality. An extrapolation of the trend shown in the chart from the research done by Iwama, Afonso and Vijayan [15] indicates that cortisol levels may take up to 60 hours to return to normal. During the period of elevated cortisol, the Koi will have a compromised immune system and may succumb to parasites, bacteria or fungus in the water. Stress also increases the permeability of the surface epithelium, including the gills, to water and ions, and thus induces systemic osmoregulation disturbances. Koi may become “stressed to death” due to hyponatremia, or low blood salts [9]. Stressors also affect the branchial structure of the gills and result in osmoregulation imbalance. The effects of such an osmotic imbalance can result in what is termed as “noninfectious dropsy”, a condition where the fish bloats. As an example, a Koi being unpacked at a Koi show was observed to be hugely bloated. The Koi was put in a .6% salt bath and returned to a more normal conformation after 4 hours. Increased permeability of the gills is one of the main reasons for the high vulnerability of fish to water pollutants. And, the more intense the stress, the more likely it is that the fish may finally be in a distressed state. The stress response in fish includes a marked increase in the oxygen uptake rate of the gills. Stress that is severe or chronic can cause harmful physical changes in the gill structure [5]. Additional stress may be fatal while the Koi is in a weakened state from a previous stressor, and as discussed earlier, stress may be thought of as cumulative. Studies have been done to determine the amount of time required for Koi to recover from a major stressor. While the evidence is not definitive because the act of drawing blood to test the Koi is, of itself, stressful and skews the results; it is thought that some of the physiological indicators may not return to normal for several months. Because there are so many uncontrollable variables in studying stress, the amount of time it takes for physiological indicators to return to normal may not be as certain as earlier research has suggested [15]. Koi do adapt to stress, but given the evidence, the authors of this paper do not recommend shipping Koi more often than every 3 months. Shipping Koi is defined as any time the Koi are bagged and boxed. Simply moving Koi within a premise does not constitute shipping, and the 3-month time period does not apply. Unless the health of the Koi is in peril, the authors of this paper would not recommend moving Koi more often than every 3 days, in order for the cortisol levels to return to normal. 1.7 General Adaptation Syndrome (GAS) Koi responses to stress are a part of a series of physiological reactions called the General Adaptation Syndrome. The term was first discussed in a paper by Selye [16] done in 1936. This syndrome is divided into three phases: the alarm reaction when stress hormones are released; a stage of resistance during which


adaptation occurs; and if the Koi cannot adapt, a stage of exhaustion followed by death. The stress response is described as being both adaptive and maladaptive [2]. For example, a primary response to stress is the release of cortisol. It makes the gills more permeable and allows for increased oxygen uptake. The fish have adapted by obtaining more oxygen from their gills when the body needs it for a fight or flight response. When the stress is severe or long lasting, the increased permeability of the gills results in electrolyte loss and increased water uptake (in freshwater fish). Thus, the stress response that was initially helpful, now becomes maladaptive, and threatens the fish’s life. Koi can easily adapt to all sizes of ponds and holding tanks as well as to a wide variety of water quality parameters. Many who have studied GAS in fish have also found that desensitization to stress reduces the physiological reactions to stress. It would be optimal to preclude the alarm reaction by having the Koi already adapted to netting and confinement before transport. Koi can easily be de-sensitized to nets. This is done in three phases, and starts by simply leaving the nets in the same place in the pond several hours a day while the Koi are being observed. This de-sensitization will probably take several weeks. Care must be taken so that all possible ways that Koi could become trapped in the net are considered and eliminated. If there is any danger of Koi becoming trapped, the nets should not be left in the pond, and nets should not be left if someone will not be there to observe the Koi and guarantee their safety. The Koi should be fed over the nets, and should be observed routinely swimming over and around the nets without reaction before advancing to the next step. Next, the nets should be slowly moved to a slightly different location on a daily basis for several weeks. The Koi should not be constrained by the nets – the purpose is to merely acquaint the Koi with net movement. Once the net movement does not cause alarm, the final phase can be initiated. The goal of this phase is to net the Koi and release them without stress. The Koi should first be slowly seined into a relatively large area such as 1/2 to 1/3 of the pond, and then immediately released. This should be repeated occasionally for several weeks, and eventually the Koi should not increase the speed of their swimming while being restricted, nor should they exhibit any other stress indicator. After several instances of calm behavior while being seined into a large area, the Koi can be further restricted to a smaller area, and then released. Once they are not agitated by being restricted to a small area, then they can be de-sensitized to a capture net. The Koi should be directed with the capture net to one corner, kept in the area for several seconds and released daily for several weeks, one Koi at a time. As the Koi adapt and are less agitated, they may be held for longer times or even bowled for a medical exam. Once Koi learn to accept netting, the lessons must be reinforced at least once a month. If the time is taken to adapt Koi to being netted, the Koi will be much less stressed by transport and even showing. During the training process, all Koi must be closely observed for disease. The goal is to adapt the Koi to stress slowly, so they do not succumb to parasites or other pathogens. If any of the Koi become ill during desensitization, or show excessive resistance to netting, such as bleeding from


the gills (a stress indicator), the desensitization process should be halted and only re-started when all the Koi have recuperated. 1.8 Parallels with Human Health Many large hospitals recognize that there is a short window of opportunity to stimulate the human immune system so that treatment is more effective. Hospital surgical ICUs amend other treatments with high doses of antioxidants for the first 3 days of care. Several Koi vets, including Dr. Sandra Yosha, DVM, PhD (see Biographies, Appendix) recommend the inclusion of an equal dosage of Vitamin C along with any antibiotic injection. Dr. Eric Johnson [17] recommends supplemental feeding of ascorbic acid (vitamin C) enhanced feeds as an ancillary treatment to all disease. The goal of the antioxidant is to stimulate the organism’s immune system so that the medicine works better. This holds for both Koi and people. Because antioxidants are routinely used in human ICUs when the patient’s immune system is extremely stressed, it makes sense to also use antioxidants when Koi are stressed. Koi stress easily, and the required handling for treatment or shipping is accompanied by even more stress. Thus it is critical to consider the effects of stress and include antioxidants as part of recuperation in any situation where Koi are handled. A good way to accomplish this is to ascertain that the feed being used is new, and has a high concentration of stabilized Vitamin C.

2.0 STRESS RESEARCH DONE FOR THIS PAPER Testing Koi for stress has always been problematic. By taking a blood sample, cortisol levels and thus stress levels can be accurately determined, but the drawing of blood is in itself extremely stressful. If the blood is drawn under anesthesia, the capture and restraint, induction and recovery from anesthesia all add to stress. If the goal of the blood draw is simply to determine cortisol levels, then blood testing may be useful. If the goal is to quantify stress or determine recovery time, a less invasive and stressful test is required to get meaningful results. A study done in 1976 by Smith and Ramos [18] proved the use of a cheap and commonly available human diagnostic test to quantify the level of stress in fish. Mullet, milkfish, bonefish and butterfly fish were tested, and it was found that all species released free hemoglobin into their skin mucus when stressed. The amount of hemoglobin in the mucus correlated to the known results derived by testing cortisol levels. The hemoglobin was detected by swiping the mucus with a Hemastix® Reagent Strip, and the change in color quantified the amount of stress. The strips look similar to pH test strips, and are made of plastic with a small pad on one end. The pad is swiped in the mucus on the fish’s back (when that section of the back is out of water), and the pad changes color to indicate the amount of stress nearly immediately. The color change is compared to a chart, and is a good indicator of the associated stress level. Smith and Ramos ascertained that the reaction in the Hemastix® was only caused by hemoglobin


in the mucus rather than by any water contaminates. The Hemastix® test is simple, quick and accurate and produces very little stress to the fish. The fish being tested must only be positioned such that an area on their back is out of water long enough to swipe a Hemastix®. Hemastix® are available over-the-counter at many pharmacies, and cost approximately $50 for 50 test strips. It was decided to use the Hemastix method to test the efficacy of the product called ULTIMATE®. The goal was to try to verify the manufacturer's claim that the product reduces stress in fish [49]. ULTIMATE® was the only product tested in association with this paper. The results of some simple testing done by Karen Wepfer Pattist indicate that the methodology suggested by Smith and Ramos does work with Koi. The number of Koi tested was not statistically significant, and no formal research paper could be written from the limited testing that was done. However, some of the general trends and observations from the testing were interesting, and perhaps worthy of more rigorous study in the future. The first test was done to ascertain that Koi would also release hemoglobin into their mucus when stressed, just as did the species in the Smith and Ramos paper. The Hemastix® clearly showed that hemoglobin was released nearly immediately when Koi are stressed, but it also showed that the test is extremely sensitive. The Hemastix® showed the maximum measurable amount of released hemoglobin with anything but the most gentle of netting and bowling. It was eventually possible to very slowly and carefully net and bowl the Koi such that no stress was detected by the Hemastix®. It was very easy to roughly net the Koi and obtain a test result of maximum measurable stress. The extreme sensitivity of the test made exact results hard to obtain, and each Koi can stress differently each time it is handled such that any results are difficult to verify. The Hemastix® reagent strips were developed to test human stool samples for minute quantities of blood. While the previous research ascertained the relationship between hemoglobin in the mucus and stress in fish, the measured changes in resulting stress level are probably more significant than a single test result in this case, due to the extremely small sample size. No Koi being sampled bled from the gills, but it is noted that blood polluted water would destroy test results. A sample of 6 large Koi and 10 small Koi were netted and bowled 5 days in a row. A log was kept of the results of each Koi. In general, it was found that large Koi stressed much less during netting and bowling than did small Koi. Some Koi always indicated higher stress, while some Koi always indicated lower stress. In another test, a Koi that had just tested low in stress was released, and more roughly netted and bowled again. The Hemastix® clearly indicated an increase in the stress level. The best way to assure that most of the Koi had the least amount of stress during netting and bowling was to move extremely slowly. It was not unusual to take up to 5 minutes to net a Koi out of a show tank in order for that Koi to indicate zero stress. It was never possible to get the lowest reading while testing Koi less than 8”. The remaining testing was done only on


large Koi that had become adapted to netting and bowling and would consistently have the lowest stress indication after being netted and bowled. Recovery time was tested next. Five larger (over 22”) Koi were stressed by chasing them with a net such that the Hemastix® test showed the highest stress level. The Koi were then put in a show tank with well-oxygenated, filtered water and very gently tested at approximately 12-hour intervals to see how long it took the Koi to indicate zero stress. 1 of the Koi indicated zero after 48 hours. 3 Koi tested zero at the 72 hour mark, and the remaining Koi still did not indicate zero stress after 96 hours. The test was repeated, except ULTIMATE® was added to the tank water. (for a discussion about ULTIMATE®, see Appendix) 6 hours after testing maximum stress, the Koi were re-tested, and 1 Koi had already reached zero stress. 2 more reached zero when tested after 12 hours, and 1 tested 0 after 24 hours. The last Koi in that test never reached the 0 level, and the testing was halted after 72 hours. It seems reasonable to say that the use of ULTIMATE® reduced stress recovery time in the majority of Koi that were tested. It is not clear which component of the ULTIMATE® was most beneficial in aiding stress recovery, but as the recovery tank had a filter that would have consumed ammonia, it may be hypothesized that the quicker recovery was based on the slime-coat enhancing properties of ULTIMATE®. Testing was then undertaken to see if the amount of time in a shipping bag increased stress. For this test, 5 larger (over 22”) Koi were very gently netted, bowled and bagged. The Koi were triple bagged alone in 40” bags, and the bags placed in regular shipping boxes. After 1 minute, the Koi were released from the bag, re-bowled and tested. All 5 Koi indicated maximum stress. Larger bag sizes and greater amounts of oxygen were tried with similar results. In fact, all Koi indicated maximum stress each time they were tested after they were left in the bag 1 minute, 1 hour, 4 hours or 8 hours. The test was repeated using ULTIMATE® as a bag additive at the rate of one teaspoon per gallon of water. All the Koi still tested maximum in stress in all but the 4 and 8 hour tests. In the 4 hour test, 3 Koi showed reduced stress. At the 8 hour test, 4 Koi showed reduced stress. From these tests, it is obvious that no matter how gently we net, bowl and bag our Koi, they will be stressed. Some Koi seem to calm down after being in the box for several hours, while others seem to continue to be highly stressed. ULTIMATE® seems to help reduce bag stress, if the Koi spend more than a few hours in the bag. Certainly, ULTIMATE® would bind the ammonia to assure that the Koi arrived without the added stress of Ammonia burns. Also, keeping the slime coat barrier in tact should have a beneficial effect by protecting the Koi from the invasion of pathogens while their immune system is compromised from the stress. There were many more tests that could have been performed. In general, the differences between individuals was at least as large as the differences derived from the testing, such that very large samples of similarly sized Koi would need to be used in order to gain reliable and significant results. While specific conclusions cannot be drawn, the existing observations would seem to indicate a beneficial effect from using ULTIMATE® in the recovery tank, and also as a bag additive. The Koi which were handled most often during the 3 months of testing


had less stress in general after being netted and bowled at the end of the testing than they did at the beginning, and had adapted quite well. Some Koi appear to rapidly acclimate to stressful conditions, while others acclimate much more slowly. Any Koi may have a previously compromised immune system, and the stressor may never be known. That is another reason why a large sample size would be needed to reduce the significance of a single, atypical result. Perhaps the most important fact that can be derived from this testing is that Koi do release hemoglobin into their slime coat extremely readily when stressed. Every aspect of handling and keeping Koi should be evaluated in an attempt to reduce stress where ever possible, especially as the additive effects of all the multiple stressors involved in shipping will have a detrimental impact on Koi. Hobbyists might find it useful to keep Hemastix® on hand and test their Koi whenever stress is suspected, but it must be noted that the stress of restraining the Koi to be tested can easily invalidate the results. It is possible to say that Hemastix® are an excellent tool for further research into the stress of Koi, and even an occasional test done by a general hobbyist could certainly be illuminating and useful in trying to learn how to handle Koi in the least stressful way possible. To achieve a zero result with the Hemastix® can certainly be considered an accomplishment and a testimonial to both the careful handling of that Koi, and the Koi’s adaptive response to handling.

3.0 PREPARATION FOR TRANSPORT 3.1 Attention to Stressors Transporting Koi involves moving them from the source water at the point of origin to the water in the transport container. At the destination, the Koi are moved again, this time from the transport container to the final destination-pond or tank. Stress and physiological stress-responses are caused by netting, handling and moving Koi, as well as by exposure to water changes [5]. Whenever Koi are stressed, cortisol is released and can be measured to indicate the severity of the stress. However, the important concept is to pay attention to the stress, not the cortisol level. Stress harms the Koi by decreasing the effectiveness of its immune system. A less than perfect immune system increases the risk of infection or other health problems to the Koi being transported, and may increase the possibility of transmitting disease to other Koi already at the destination [19]. Transferring fish from pond to pond is a common way to spread disease and parasites. Add to that equation stressed fish with suppressed immune systems that are highly susceptible to disease. 3.2 Fasting Koi are poikilothermic, and that makes them different from the other animals we keep as pets. The old term was “cold-blooded”, but that term has been outdated (see Glossary for further discussion). “Warm-blooded” animals, including humans, dogs and cats, maintain a consistent body temperature


regardless of the surroundings. Poikilotherms are different, because their metabolism is tied to the temperature of their environment [20]. Koi can survive water temperatures from about 35 to 90 [21]. In cool water temperatures, a Koi’s metabolism slows down and they appear to go dormant. That condition is correctly called torpor. Torpor is an endogenous suppression and activation of metabolic processes creating a dormant state where dormancy is interrupted with periodic arousals, indicating that torpor is not temperature driven. During the periods of arousal, fish will eat flora and fauna living on the bottom of the pond. When the Koi's metabolism is slowed, food takes longer to digest, and medicine takes longer to disperse and be circulated in their bodies. Warm-blooded animals cannot go without food for several weeks without harmful consequences, but Koi can. During winter, Koi in some areas of the country do not eat food fed by humans for over 6 months (most experts recommend not feeding Koi when the water temperature is below 50 degrees). Koi survive these long periods in cold water because their reduced metabolism requires less food, and the required food can be acquired by eating algae from the sides and bottoms of the pond. Even when the water is warmer, Koi are not harmed by fasting for a few weeks [23]. According to Stoskopf [6] “The most common trouble with fish during a vacation is overfeeding by an inexperienced and overzealous friend or neighbor.” Before shipment, Koi should be fasted for a week so that all food clears their systems. It is important that Koi owners understand that it is not appropriate to assume that our discomfort from a week of hunger would be the same for our poikilothermic pets [23]. Fasting prior to transport clears the intestine and thus reduces the possible pollution in the transport water [19]. The fasting/holding period needs to be long enough for the food to clear the gut and for the fasting to have its effect on the metabolism, i.e. the amount of ammonia released into the blood stream is reduced. This takes a few days. A Koi’s metabolism increases with feeding and digestion, resulting in increased oxygen demand [24]. Koi metabolism slows down during fasting, so fasted Koi will consume less oxygen during transport. Koi are evolutionarily adapted to intermittent feeding and the speed at which food clears the Koi’s gut is affected by water temperature and stress, and therefore it may take considerable time for fasting to produce a physiological effect. A study done by Phillips and Brockway [25] showed that using cooler transport water and fasting prior to transport had double benefits. First, less ammonia was released into the transport water and secondly, less oxygen was consumed by the fish during transport. While this study was done on trout, we can apply the same principles to Koi and many other fish being transported. If Koi are in ponds prior to transport, they will continuously consume algae in the absence of other food. A total fast can only be accomplished in a clean holding tank. Koi stress when held alone, so at least 2 Koi should always be kept in a holding tank, even if the extra Koi will not be transported [6]. 3.3 Holding Tank Considerations It is always critical to weigh alternatives when handling Koi. If Koi are being transported to a Koi Show, a quarantine tank will be required for the returning


Koi, and it is just as easy to set up the tank before the show and use it as a holding tank to fast the Koi. If the Koi are going on a one-way journey, it may be easier to leave the Koi in the pond and fast the pond, understanding that a total fast will not be achieved. If the proposed journey is shorter than 6 hours and ULTIMATE® is used as a bag additive, the bag pollution caused by the algae in the gut should be less of a problem. The short duration of the transport and the use of ULTIMATE® should mitigate the dangers of not achieving a full fast. 3.4 Holding Tank Optimally, Koi would be moved to a clean holding tank at least 1 week before transport. In the holding tank, Koi can be watched, fasted and allowed to recover from the stress of being captured and removed from the pond. The rest of the Koi in the pond may continue to consume a regular diet. Temperature, pH and total alkalinity in the holding water should be kept stable. The water should be tested daily, and baking soda and/or calcium may be added as needed. (see Appendix for a discussion on “Maintaining a Stable pH and Calcium” in a holding or quarantine tank) Koi should have adequate water volume so that they do not experience stress from crowding, and at least 100 gallons for each Koi over 15” is recommended [26]. A more accurate system would be to use the tank-loading formula developed to maintain good water quality at Koi Shows. (see Table #5, Appendix, for Tank Loading formula) Fish crowding is the behavioral requirement of fish for physical space, and is different from fish loading which is the carrying capacity of the tank water in terms of oxygen availability and toxin build-up. Both crowding and loading need to be considered when choosing a holding tank. There should be sufficient aeration to reach oxygen saturation at the given water temperature (see Table #2, Appendix). ULTIMATE®® should be used in the water, and will serve several purposes. First, it will neutralize any chlorine or chloramines from the supply water and it will bind all ammonia produced by the Koi. Secondly, it will replace any slime coat that may have been removed when the Koi was netted and handled, and third, it will bind heavy metals. Holding tank water changes should optimally be achieved by a slow, continuous trickle of fresh water rather than more stressful major water changes. Well water that contains no chlorine or chloramines should be used if possible. If the incoming water contains chlorine or chloramines, additional ULTIMATE® should be added at each water change to treat the volume of water changed. The total daily dose of ULTIMATE® will be determined by the amount of water changed and the number of Koi in quarantine. If ULTIMATE® is being used to both detoxify ammonia and to treat incoming water; an additional dosage may be needed if the combined ammonia, chlorine or chloramines present exceed 3 ppm [49]. (for product dosages, see Appendix for the ULTIMATE® Product Data Sheet) The tank should have filtration that is suitable for a fish habitat of that water volume. ULTIMATE® should not be used as a substitute for a working filter.


Adding Calcium to the water during both the holding and quarantine period may help stressed Koi [13, 48]. This requires knowledge of the calcium ion content of the water. Adding calcium to calcium rich water produces no benefits. Adding calcium too quickly to water with very low hardness can produce significant pH changes, which may irritate the Koi, but this is only a concern in very-low-hardness water, and is more of an issue if calcium hydroxide is used than if Calcium Chloride is used. Calcium is an electrolyte and reduces the permeability of the gills to water, thus reducing water and electrolytic flux [7]. The gills of stressed Koi have increased permeability due to the cortisol effect and up to about 80mg/L calcium can be used to offset the increased permeability. Again, all factors should be considered before adding anything to the water, but it should be understood that any reduction in oxygen uptake as a result of adding calcium is relatively unimportant compared to the dropsy and osmotic shock that calcium helps to prevent. Adding calcium chloride to water does not release chlorine ions which would have to be neutralized with de-chlorinator. (see notes, Table #4, Appendix) Note that calcium ion content is not the same as alkalinity (KH), or hardness (GH). The test for hardness (GH) does not distinguish which ions are causing the hardness. There are calcium ion specific test kits on the market manufactured by Hagen (sold under the Nutrifin label), Salifert, Aquarium Pharmaceuticals, and Red Sea Fish Pharmaceuticals, and others that can be used to easily and accurately determine the calcium ion content of water. 3.5 Attention to Slime Coat The primary defense that Koi have against pathogens is their slime coat. The slime coat or cuticle is part of the epidermis layer of the Koi’s skin. It contains natural antibiotics and provides a barrier to parasites and bacteria. Netting and bagging Koi in even the most careful way is bound to strip some slime coat. If there is a small gap in the slime coat and pathogens are present, Koi may become infected. While that infection is usually handled by the Koi’s immune system, if the immune system is weakened by stress, the probability of infection greatly increases. Small amounts of salt, such as the .3% that is recommended for use in transport bags and isolation tanks, also has the effect of increasing slime coat. This works because salt is a mild irritant which stimulates mucus secretion [50, 51, 52]. However, large amounts of salt, such as 1-3% that may be used as baths or dips, will strip slime coat [53], and would kill Koi in any prolonged exposure. A case can be made to enhance slime-coat and thus close the portal of entry for pathogens whenever Koi are handled. Older chemicals marketed for the purpose of increasing slime coat in Koi functioned by irritating the Koi into producing more slime. Not only did being exposed to the chemical irritants add stress to the Koi, but the greatly increased slime also coated the gills and stressed the Koi further by reducing their ability to extract oxygen from the water. Koi often suffocated, or quickly succumbed to pathogens, and the older slime-coat enhancing products fell from favor. Current technology has produced a product called “ULTIMATE®,” a polymer-based product that, according to the


product literature, preferentially replaces slime coat only where it is missing, and does not coat the gills [49]. If the portal of entry through the skin is closed and the Koi has an unbroken layer of slime coat, a Koi is much less likely to succumb to pathogens even if it is exposed when its immune system is challenged. The instantaneous replacement of the slime coat due to the polymer is in the water means that the primary portal of entry is always closed. The authors of this paper therefore suggest that ULTIMATE® be used before, during and after transport to minimize the likelihood that the stress of transport will later cause disease. Before Transport - ULTIMATE® is recommended while the Koi are being fasted before shipment During Transport - ULTIMATE® is recommended as a bag additive After Transport - ULTIMATE® is recommended as a recuperative aid for the period that Koi are quarantined after shipment. 3.6 ULTIMATE® and Gills To check the claim that ULTIMATE® does not coat gills [49], one of the authors of this study, Karen Wepfer Pattist, did a simple test. 6 Koi were anesthetized and gill scraped, and the slides observed under a microscope to check the level of mucus. These 6 Koi were then placed in a tank containing ULTIMATE® at 5x the dosage recommended on the label. After 24 hours, each of the Koi was again anesthetized and gill scraped, and no increase in mucus was observed under the microscope. The test was repeated after 48 hours and again after 72 hours with the same result. As a control, another Koi anesthetized and was placed in water containing another over-the-counter stress product. When that Koi was again anesthetized and tested after 24 hours, there was an estimated 12-fold increase in mucus. While the sample size was not statistically significant, and while the observations were subjective, the observer was a trained Koi Health Advisor that has seen many slides with excessive mucus secretions. There was no observable change in the amount of mucus in any of the slides from the Koi kept in ULTIMATE® as seen under the microscope. These results would tend to corroborate the claim that ULTIMATE® and Stress-X® (the component of ULTIMATE® responsible for replacing slime coat) does not tend to suffocate the Koi with excess mucus in the gills. (see Appendix for a more detailed discussion of ULTIMATE®)

4.0 TRANSPORT CONSIDERATIONS Next, the physiological responses of Koi to all aspects of transport will be considered, and recommended preparations and procedures for transport will be explained. 4.1 Minimizing Stress While Catching Koi


The use of a holding tank to fast Koi means that the Koi do not have to be moved until assistance is available, which allows the move to take place in an orderly fashion with reduced stress to the handlers as well as the Koi. The Koi can be leisurely moved to the holding tank in advance of transport, and then easily bagged from the tank. It is imperative that stress should be minimized as Koi are netted, bowled and bagged. Any handling of Koi is a stressor [14]. The key to minimizing stress is for all the people involved to move slowly. A Koi’s fight-or-flight response [26] will be triggered by any quick movements. While netting Koi, a slow and persistent demeanor is always preferable to any fast movement. Five minutes may seem forever to have to slowly move a net in order to get a Koi into the bowl, but taking five minutes to approach the Koi slowly and let them get used to the net is always faster than netting scared Koi. Once Koi are scared, they will swim very rapidly and it often takes more than 30 minutes for Koi to slow down enough to be captured. The frightened Koi will be stressed. Stressed Koi bleed more easily out of their gills, and any blood will instantly pollute the shipping water or holding tank water. Stress is easily observed in Koi with white in their pattern, as the stress brings capillaries to the surface, and the Koi may appear to blush (turn “pink”) or have discernable red veining in the white. The red muscle lies closest to the skin and has the most veins in it. Veining in the white muscle area may not be as visible, but does not necessarily indicate the absence of stress. “Pink” Koi are the result of some stressor, and can often be observed at Koi shows, and pink skin shows an obvious stress-related problem. Certainly, any Koi may be stressed without necessarily showing it and this symptom may be harder to see in Koi with background colors other than white. It is obvious that “pink” Koi have been stressed, and while the stressor may have been unavoidable, a “pink” Koi will not be judged as highly as it could have been. According to head judge Burt Ballou (see Biographies, Appendix), if the U.S. judge can’t tell what the skin quality is because of the “pinkness,” or if there are 2 Koi with comparable skin quality and all other factors are equal except that one is “pink,” the Koi with the “pink” skin will be judged as inferior. In Japan, “pink” skin is always judged as a fault, because showing Koi is thought to require the knowledge of how to make Koi look their very best, and “pink” skin shows an obvious stress-related problem. When bagging Koi, a new bag should always be used to avoid contamination. Koi should always be transported with a minimum of 2 bags, both to help maintain water temperature and as a safety precaution against leaks. A re-used bag may be employed as the second or outer bag, if it is desirable to save money. Some people prefer to move Koi from the bowl to the bag with a sock net. Using a sock net is another opportunity for stress. Only sock nets that hold water should be used. Sock nets that do not hold water will strip the slime coat at exactly the time when the Koi need all of their defenses to counteract the effects of stress. Even a sock net that does hold water will agitate the Koi, and must be well supported so that the spine of the Koi remains straight while the Koi is being


moved. Sock nets are designed to provide the fish a one-way travel path through the net. Koi should always enter a sock net nose first in order to minimize the danger of tearing fins during release. Another technique is to direct Koi to the handlers with a pole net, and then scoop them up using a transport bag directly from the water. A sock net could be substituted for the bag, but is not recommended as it introduces another handling step. Bagging directly from the tank is impractical unless the tank water is being used as transport water, and even if tank water is to be used, it is difficult to capture the right amount of water in the bag along with the Koi without agitating the Koi. This technique can panic skittish Koi, and must be performed slowly and gently. In general, the least stressful and safest system is to bag Koi from a bowl. It should be noted that spooked Koi are likely to try and jump from a bowl, and the bowl may be covered with a net for several minutes to see if the Koi will calm down before it is handled. If the bag water is to be different from the source water, the Koi may be lifted by hand into the bag. This is done more safely while the bowl is still floating in water, such that if the Koi jumps or is dropped, it lands in the water and the chance of injury is reduced. Choices for moving Koi to bags or transport containers:

• Bowl the Koi, and lift the Koi by hand directly from the floating bowl into the bag or transport container. This takes skill and ability, but if the Koi is dropped, it is likely to land back in the water.

• Bowl the Koi, then move the covered bowl out of the water to a location where access is better and lifting is easier, and then lift the Koi into the bag or transport container. This makes the lifting and moving easier, but if the Koi is dropped, it is likely to land on the ground.

• Bowl the Koi, then use a sock net to transfer the Koi to the bag or transport container. This will mean that at least some of the bowl water is used in transport, will add the additional stress of another step to the transport experience, and will remove some slime coat from the Koi.

• Bowl the Koi, and then bag the Koi directly from the bowl using the water in the bowl as transport water. This can be the least stressful to the Koi, but the water in the bowl needs to be suitable transport water. This system does not allow additives to be put into the bag water in advance.

• Use a pole net to direct the Koi, and then bag the Koi directly from the tank. This technique should only be used if tank water is to be used as transport water. This system does not allow additives to be put into the bag water in advance.

Things to consider:

How skilled and physically capable is the person that will lift and move the Koi by hand?

How much assistance is available to help lift and move the Koi and/or the bowl?

Is there a concern about dropping the Koi? Should the source water be used in the transport container?

4.2 Quick and Efficient Transport


Koi should be transported quickly and efficiently for two reasons. First, stress is minimized, and second, the physical harm inherent to the time the Koi are exposed to the contaminated water during transport is minimized. It should be noted that only the efficient transport of bagged Koi is being discussed, and speed in capturing and bagging Koi is not being advocated. This quickness and efficiency of transport requires proper preparation and planning. In order to make the transportation of Koi as quick and efficient as possible, plan for the following:

• Have all the proper Koi handling and transportation equipment ready. • Have enough assistance at each stage where the Koi are being handled. • Have the necessary water testing equipment ready, so that the water can be

tested for the critical parameters at each stage of the move. o Test the source water for Ammonia, Nitrite, pH, Alkalinity and

Temperature. o Test the transport water (if not using the source water) and the

destination water for pH, Alkalinity and Temperature. o Be prepared to adjust the pH/Alkalinity with baking soda, and the

temperature with heat or ice packs. • Know as much as possible about the transport effort at the receiving end.

Where is the fish going, how is it going to be handled? By whom? Water considerations? Water parameters??

The possibility of transportation delays must be considered. The water temperature should remain stabile and dissolved oxygen should be sufficient for the entire time that the Koi are bagged, regardless of reasonable delays. A sufficient supply of oxygen is even more important for larger Koi, as they will succumb to low oxygen long before smaller Koi [6]. It is always recommended to ship the Koi in good water and as much pure oxygen as possible. Stoskopf [6] says that “…oxygen is much more important than water, and there should be twice as much air space in the sealed bag as water space.” Koi are often shipped together in bags from dealers or farms to minimize shipping costs. In order to maximize the health of the Koi, it is recommended to ship each Koi in a separate bag. The use of bags smaller than approximately 20 x 40” is not advocated. Koi over 18” should be triple bagged. The bags should be placed in boxes that are lined with Styrofoam or other insulation on all sides. Many Koi farms suggest using coolers to maintain a more even temperature (instead of boxes) for valuable Koi. (A list of breeders and dealers interviewed can be found in Table #3, Appendix) 4.3 Organizing to Transport Koi The most important item needed for transporting Koi is organization. Before the Koi are to be handled, everything must be prepared. The need to keep the Koi in a holding tank to which ULTIMATE® has been added has already been discussed. The Koi are fasted during that time so that homeostasis returns to normal after the stress of being netted from the pond, and food clears their digestive system. Koi travel best at cooler temperatures. The optimal shipping temperature is around 45 degrees, according to a majority of Koi farms


interviewed in Table #3. During the holding/fasting period, ice can be used to lower the tank temperature slowly, by no more than 2 degrees in 12 hours. This takes constant vigilance, and if not carefully monitored, more harm than good can be done. If the temperature varies up and down rather than a gradual consistent decrease, the stress induced will compromise the Koi’s immune system before the more stressful event of being bagged and boxed. Before any nets enter the water, all bags and boxes should be prepared. Ice packs should be wrapped, to insulate the Koi from trauma and to stretch the time that the ice lasts, and should be waiting in the box. The goal of adding ice packs is to air-condition the container to maintain the existing water temperature for the entire time the Koi are in the box/cooler. The wrapped ice packs should be duct-taped to the sides or top of the box such that they cannot come into direct contact with the Koi bags. Koi bags should be doubled up (tripled for Koi over 18”), and transport water should be added to the inner bag. The quantity of water should be just sufficient to cover the Koi’s gills when the bag is at rest in the container. Thus the amount of water needed will depend on the size of the bag, the size of the Koi and the relative position of the bag in the container. To facilitate placing one Koi bag inside another, it is often helpful to place a small amount of water into the outside bag to hold it open while the next inner bag is inserted. ULTIMATE® and salt should be added to the shipping water before the Koi are lifted into the bags. (see note at end of section 5.11) Salt must be completely dissolved before Koi are added, as contact with un-dissolved salt may cause burns. If Koi are added to the bag along with water, the bag water should be tested for salinity and dissolved salt should be added as required to regain a concentration of .3%. The ULTIMATE® dose should be sufficient to neutralize at least double the expected amount of ammonia that will be produced by the Koi during transport, in order to allow for delays. The dosage must also account for any chlorine or chloramines that must be neutralized in the shipping water. A "dose" of ULTIMATE® is defined as 1 teaspoon per 10 gallons of water. That "dose" will neutralize 3 ppm of ammonia, chorine and chloramines in total [49]. For example, if the shipping water contains 3 ppm chlorine, and if it is known that ammonia will be released by the Koi during shipment, then at least a double dose of ULTIMATE® should be used. It should be noted that it is impossible to overdose ULTIMATE®, and that Koi can survive in pure ULTIMATE®. (see Appendix for more about ULTIMATE®) Shipping bags should be prepared by rolling down the top edge to make a cuff. The cuff will help keep the bags open while the Koi are being moved. Once all the containers are prepared, the last job is to net the Koi. As each Koi is carefully placed into its bag, the cuff can gently and slowly be straightened. Moving slowly will help keep the Koi calm and reduce stress. When the bag is filled with the desired amount of oxygen, seal the bag with bands, and place Styrofoam on top of the bag. Lastly the box is closed and sealed. If there is a shortage of hands, it may be easier to prepare all the bags and boxes at once like an assembly line. If all the Koi are to be bagged before oxygen is added, place a few twists in the neck of the bags once the Koi is added, and then the


twisted bag neck can be temporarily wedged between 2 flaps of the box. It is not recommended to leave the bags open once the Koi are inside, as the Koi may jump. Leave plenty of air in the bags while working on other Koi. Ewy [19] suggests that Koi should be transported in a dark environment, so several sheets of newspaper should be used to cover a bag containing Koi until the box is properly sealed. Bagged Koi must be kept in the shade. (see Section 6, Equipment and Supplies, for a thorough discussion of each phase of bagging and boxing) 4.4 Bag/Transport Tank Water Since Koi are acclimated to the water at the point of origin, it is common practice to use the source water as transport water to eliminate the additional stress of adjusting to different water quality. The choice of transport water is critical. Even fresh, clean water can have a different pH or temperature from the pond water, and Koi stress from differences in water parameters [11, 13, 26]. Municipal water may contain chlorine or chloramines. Well water should be heavily aerated for several hours or even days to boost the oxygen and remove harmful gasses such as carbon dioxide and hydrogen sulfide, and to prevent gas supersaturation [27]. Any water may contain harmful levels of heavy metals. If the water quality in the originating holding-pond or tank is excellent, then it is the preferred choice to be used for transport. If the water quality is less than optimal, the issues of the original water should be weighed against the problems of the available fresh water, and a decision should be made that minimizes detrimental effects. 4.5 Un-bagging Koi The primary goals while un-bagging the Koi at their destination should be to: 1) Remove them quickly and safely from the polluted bag water 2) Make sure that as little-as-possible bag water ends up in the destination water. The bag water will be extremely polluted by ammonia and carbon dioxide. The water will be low in pH and high in organic material. Koi should be removed from the bag as quickly as possible, and the bag water should be properly disposed of. The bag with the Koi should be floated for 15 to 30 minutes to help reduce significant temperature differences between the bag and destination water. This must be done in the shade, or the Koi can literally poach in the extremely high heat that quickly builds up inside a bag left in the sun. Koi often stress and become quite agitated when the bag is removed from the box. Again, one must weigh the dangers between the damage inflicted by an extremely agitated Koi in a bag of polluted water against the shock caused by prematurely releasing the Koi. An agitated Koi can and will damage fins by thrashing in a bag. There is also the possibility that a broken fin ray may puncture the bag itself. But, releasing Koi into water that is more than 5 degrees different may shock them [20], especially warmer to cooler, and will certainly harm their immune system [26]. A variety of sudden changes in a Koi’s environment can trigger a stress reaction, but various genetic, developmental, physical, nutritional and environmental factors can have a modifying effect on the magnitude and duration of the stress response [2].


Koi should significantly calm down within 5 minutes while floating in the bag. If they are still highly agitated after 5 minutes, they should probably be removed from the bag despite the detrimental effects of temperature shock, if the water is within 10 degrees. Once the temperature has equalized between the bag and the destination water, the Koi should safely be removed from the bag. It is not recommended to open the bag and add destination water into the bag. Instead, the Koi should be removed immediately. Moving the Koi safely is the primary concern. It is always fastest and less stressful to the Koi to cut the bags open with a sharp scissors, being careful not to nick the Koi. Never re-use any bag that has had Koi in it. Even if the bags are not cut open, always cut the bands with a sharp scissors or diagonal clipper. Never try and remove the bands so they can be reused, as the activity required will agitate and stress the Koi. Once the bands are removed, the top of the bags should be gently rolled into a cuff, down to nearly the level of the water. The bag may be held by an assistant to keep from tipping. At this point, an experienced Koi handler will reach into the bag and gently lift the Koi over the bag edge and place it into a bowl for examination and photography or directly into the destination water. The bag and all the polluted water should be lifted from the water and safely disposed of. Care should be taken to find an adequate location to dispose of the polluted water (which may contain chemicals and salt). The bag water should not be allowed to run off into area waterways. Some Koi Shows require bag water to be dumped into a holding tank and disinfected before disposal. Check for any local requirements before disposing of the bag water. No Koi Show allows the Koi owner to add any chemical or substance, even salt, to the water in their show tanks without permission from the Show Chairperson. If the bag is floating in an easily accessible tank where the Koi will be released, and there is no danger of dropping the Koi into anything but water, the Koi should be lifted by hand, as this method will remove the least amount of slime coat and is less stressful to the Koi because it can be done quickly. The Koi may be poured into a bowl along with the bag water, and then lifted into the destination water. While using the bowl may help prevent bag water from polluting the destination water, there is the added danger of the Koi hitting the bowl if it is dropped. If the Koi must be lifted over hard ground and there are no experienced Koi wranglers present, several other options may be preferable to the risk of dropping the Koi. Once the bag is cut open, squeeze an opposite corner of the bag to protect the Koi from being cut, and clip off the small, protruding corner of the bag with a scissors. Drain some of the water from the bag, and use the bag as a sock net to move the Koi. Be sure to support the Koi by holding both ends of the bag such that the Koi’s spine remains straight. When the destination is reached, place the bag in the water and allow the Koi to exit from the open end (not the end with the cut corner). This system is preferable to dropping the Koi, but will remove some slime coat, may cause fin damage, and will allow some polluted shipping water to contaminate the destination water. The Koi should optimally leave the bag facing the open end, but that may not always be possible. Koi often become severely agitated as the water is drained from the bag, and that stress will add to the existing multiple stresses of netting, bagging and transport.


Koi should never be moved with a net. Koi that are moved in a net may sustain trauma through either spinal or internal injuries, and should always be moved by hand or other methods that will guarantee that their spines are kept straight, and their weight is well supported. (see Appendix for web sites with articles about moving large Koi) Using a sock net to dip into the bag and move the Koi is another alternative. It will also strip slime coat and will carry at least some of the polluted bag water into the destination water. Often, the system which causes the least stress to the humans involved will also end up causing the least stress to the Koi. All these options should be considered and a plan formed before the bag is touched. When Koi are removed from a bag, they may exhibit shock by suddenly becoming very still. At this point, they should be rested in excellent water with ULTIMATE® added. At a Koi show, stress-reducing chemicals will already be in the water, and the Koi should recover well, and be swimming normally within an hour. It is not unusual to observe Koi huddled together on the bottom in a dark corner of a show tank. These Koi are clearly uncomfortable with their surroundings, and may be exhibiting behavior that indicates they could have come from darker or more opaque water, and feel very vulnerable in the clean shallow tank water at the show. Learning to observe Koi’s normal behavior in their home pond will provide one of the best indicators that they are exhibiting abnormal behavior because something is wrong. The Koi could be in shock or severely stressed from the transport experience or from the differences between their home water and the destination water. When ever Koi are observed sitting on the bottom, especially with their pectoral fins clamped to their sides, Koi keepers should question the cause of the distress and take steps to minimize it. 4.6 The Recovery and Quarantine Tank The recovery or quarantine tank is the final step in the Koi transport process. This research report will only cover some brief details here. Most of the same concepts apply equally to new arrivals at a location as to Koi returning from a show, exhibition or veterinary surgery. When Koi are being moved within a given location, such as moving fish from indoors to outdoors or from pond to pond, the transport time and stress may be minimal and there may be no need for recovery or quarantine. The reasons to use a recovery tank are somewhat different than the reasons for quarantine. In recovery, stress is reduced so that fish can regain homeostasis (osmotic and acid-base balances) and adapt to new water parameters. This allows the immune functions and feeding patterns to return to normal. In quarantine, the fish are isolated so that care takers can evaluate the health and the condition of the fish and treat any disease or illness. This is easier to do in a tank isolated from other fish populations, and it prevents the spread of pathogens to those other populations. A working filter or ULTIMATE® should be used to assure the Koi are not additionally stressed from ammonia in the tank. While flow-through is an option for other Koi keeping scenarios, it has too many bio-security risks to be used in any situation when Koi are being isolated.


The recommendations for the management of water quality, control over fish crowding and the use of ULTIMATE® apply equally to recovery tanks, quarantine tanks and holding tanks. Refer to the Holding Tank section for details. For quarantine tanks, time and temperature are also important. Cover [28], the head of Project KHV for the Associated Koi Clubs of America, recommends:

“Quarantine all new fish, and those returning to the facility, at 70° F to 75° F for at least three weeks.”

Griffith recommends a quarantine time of 4 weeks [29] at 75 degrees F [30]. Temperatures may vary between 70 and 80 degrees, but if the water temperature varies outside of that range, it is recommended to start the 4 weeks again so that the entire quarantine period maintains 70 – 80 degrees F [30]. The 4 week time period allows for complete treatment of any parasites because the parasites will have gone through one entire life cycle. 3-4 weeks at 75 degrees will also allow for the detection of fish infected with viruses like Koi Herpes Virus [30, 31]. (for a list of web sites with information on quarantine, see Appendix) At the end of the isolation period, a pond owner may take the opportunity to add a Koi from their existing pond collection into the isolation tank. New Koi may introduce different gut flora to the pond, and while those bacteria may not be harmful in the long run, they may cause the existing pond inhabitants to go through a period of adaptation where they appear lethargic. The existing Koi may have the same effect on the new Koi, so a period of observation and adaptation may be a good reason to extend the isolation until both the new and existing Koi are behaving normally. In general, Koi being held in recovery and quarantine tanks should be fed very lightly. Use a fresh, high-protein, high-quality complete Koi food, and offer no more than a few pellets to each fish twice a day. The goals are to both keep the fish eating and interested in food, and to minimize waste. The pellet size should be small enough that the smallest fish can easily crush several of the pellets at the same time. Commercial Koi food pellets are hard and can be quite sharp. Larger sizes can cause sores in the Koi’s mouth, as Koi are rarely willing to delay consumption long enough for the pellets to soften in the water. If a large pellet size is the only option available, pre-soften larger pellets by placing them in water for 2-3 minutes before feeding [32]. The time required to pre-soften food varies with the manufacturer, and close observation will be required because some food can go from hard to disintegrated within a few minutes. In a pond, Koi may be fed small quantities of fresh fruits and vegetables as a supplement to their pelletized staple diet [20]. This is not recommended practice for a quarantine or recovery tank because these foods produce a higher quantity of waste which can quickly degrade water quality. (see Appendix for more on Koi food) 4.7 Transport Injuries An important factor to consider when transporting Koi is the correlation between stressful events such as injuries, and how injuries may increase susceptibility to pathogens later on. Physical injuries could include split or broken fins, swelling from bruising, penetrating wounds and loss of scales. Wounds can occur from


netting, jumping and hitting sharp objects, rough handling, stressing in the bag or transporting incorrectly. Being prepared and using the proper tools may prevent injury and will decrease the severity of injuries. Never lift Koi out of water with a regular net. Koi grow to be quite heavy fish for their size and their spines need the support of the water. Lifting Koi with a net can cause a spinal cord injury, damaged scales, loss of slime coat and split fins. If Koi jump onto the ground or hit the side of the pond, very similar damages can occur. Traumatic injuries include those that are noticeable on the body or fins. If the Koi becomes injured in any way, it should be bowled to examine the extent of injuries and to facilitate treatment. That Koi should be transported at a later time, after all injuries are healed and the Koi is less stressed. If a Koi has jumped and impacted a hard surface, rinse the Koi off well with water, and check the gills to be sure they are free of dirt. Any close-by water source may be used for a quick rinse, if necessary. Never rinse gills with anything other than water. If the Koi has been knocked unconscious, support them gently with both hands, and aid them to swim slowly forward in a circle to revive them. The Koi may be held over an airstone instead. To avoid damaging their fragile gills, Koi should always be helped to swim in a normal, forward motion. However, if the Koi has been out of water for some time, check the gills to see if they are stuck together. If the gills are stuck together, then a gentle backwards movement with the operculum open is warranted. If the Koi do not recover within half an hour, place them in a closed bag containing good water, pure oxygen, 0.3% salt that has been completely dissolved (see Table #4, Appendix), and 1/2 cup of ULTIMATE®. Float the bag in the shade until the Koi recovers, which may take up to 24 hours. If there is swelling on the brain, the Koi will most likely not recover. Swelling can be caused by either bruising (concussion) or the lack of oxygen being received by the brain (anoxia) when a fish is out of water. Anoxia interrupts cell function including control of cell volume. As cell volumes increase, there is also an increase in intracranial pressure. When this pressure exceeds the blood pressure, blood flow to the brain will stop (a state called ischemia). At this stage, the brain cannot be saved even if the organism is re-oxygenated [33]. A Koi’s heart may continue to beat even after an hour out of water, but if blood is not flowing to the brain, the fish will die. In some cases, swelling may be effectively reduced by an injection of the steroid Dexamethasone. Dexamethasone is also used to treat Koi that have swelling from cold and electrical shock [34]. (information on Dexamethasone can be found in Section 6.10, Emergency Kit) Jumping may also lead to injuries such as torn fins, missing scales or blunt trauma wounds. If a Koi has any injury, it should not be transported. Treatments themselves may strip additional slime coat, so any recuperation should be done with ULTIMATE® in the water to help replace slime coat. Temporary treatments for an injured area could include sedating the Koi and cleaning the area with Iodine or hydrogen peroxide, making absolutely certain that neither substance gets in the eyes, nose, gills or mouth. A topical antibiotic can be applied over a


wound [35]. Whenever Koi suffer damaged skin or loss of slime coat, their bodies will have to work harder to prevent the influx of water from their environment. Koi’s blood is about .9% salt, just like human blood. Because water tends to move to areas where the concentration of salt is higher, Koi must continuously battle becoming over-hydrated. Salting the water to 0.3% will lessen the gradient that the Koi’s body must work against to resist the uptake of water, and will reduce osmotic stress. Care should be taken to completely dissolve salt before adding Koi, as direct contact with salt may burn Koi skin. The salt will also work to stimulate slime coat production [50, 51, 52]. The optimal temperature for a Koi’s immune system is 74-76 degrees [21, 36], and slowly raising the temperature of a quarantine/sick tank to near 75 degrees will help Koi to heal quickly. Keep a close watch over the Koi for the next few days in a quarantine or sick tank. Observe the healing of the injuries, and check to see if the injured Koi is isolated itself, as this can be a sign that they are getting worse. Having the Koi in a quarantine/sick tank is an opportune time to scrape the Koi, and check for parasites using a microscope. If the Koi is not healing well, check for stressors, including the presence of parasites. All stressors will impede the immune system and prevent the Koi from healing. Parasites can be much more easily treated in the smaller volume of water in a quarantine/sick tank than in a pond. Continue to check for infection around the wound site until it has healed over with new white skin. If any redness develops around the wound, Antibiotic food may be used according to package directions. To avoid resistance, it is generally suggested that antibiotic food be used no longer than a two weeks. If the Koi have stopped eating, sometimes freeze dried krill can be fed to entice them to eat. Split fins or broken rays can occur from overcrowding in the transport bag, from a sudden stop or from transporting incorrectly. Small splits in fins often heal on their own, and younger Koi tend to heal faster than older Koi. Feeding nutritional food, keeping perfect water and time are usually all that are needed for Koi to recover and heal from fin injuries. A fin that is split all the way down to the body may not heal without intervention. Seek professional help to repair the fin.

5.0 TRANSPORT ADDITIVES The water in the transport bag or tank can be chemically treated by the use of additives to help achieve optimal water quality. Different additives are formulated to accomplish various results. Bag Additives should be added to the water before the Koi is placed into the bag. Bag additives can turn out to be more harmful than helpful. One thing is certain – the water chemistry in a transport bag or tank is constantly changing. It is important to examine the goal of each of the additives as it pertains to transporting Koi.


Oxygen, while the most important item added to the bag, is not counted as an additive. It is important to remember that if the Koi are agitated and stressed during shipment, they will breathe faster and use up the available oxygen in the water more quickly. Excessive movement will dislodge slime coat which will coat the Koi’s gills and limit oxygen uptake. 5.1 Detoxification of Ammonia Koi are constantly producing ammonia, even at rest, and even when fasted.

“It is important to know that 75% of the ammonia secreted by the fish originates from the gills, and only 25% from urine and excrements. This implies that fish constantly secrete ammonium, even when they are not fed (e.g. during wintertime) [37].”

This ammonia builds up to toxic levels quickly in the confines of a transport bag. Thus, even perfectly ‘aged and aerated’ well water will eventually kill Koi if the Koi are left in the bag too long. As stated before, ammonia is less toxic at lower pH and in cooler water. (see Table #1, Appendix) Any additive that raises pH will actually increase ammonia toxicity. Any additive that detoxifies Chloramines, without also binding the released Ammonia, may increase the toxicity of the bag water. Ammonia toxicity depends on other factors as well, including the health of the Koi’s gills, their respiration rate, the temperature of the water and the salinity of the water. If the Koi’s gills are compromised before shipping, ammonia in the transport bag will continue to damage the gills, and it will take less ammonia to do more damage. Ammonia causes burning to the secondary lamellae where oxygen is transferred. 5.2 Detoxification of Chlorine and Chloramines Either Chloramines (“combined available chlorine”) or Chlorine (“free available chlorine” or hypochlorite, OCl-) is used by municipal water supplies to disinfect drinking water, making it safe for human consumption. Only water safe for Koi should be used in transport, and thus it is assumed that municipal water containing chlorine or chloramines must be treated. If Chlorine alone needs to be neutralized, then Sodium Thiosulfate can be used. Any time Sodium Thiosulfate is used, the water should be tested for residual Chlorine by means of a Chlorine test kit after all the Sodium Thiosulfate has dissolved and the water has been circulated. If Chloramines are used by the water supply and if that water is treated as if it contained only Chlorine, one chemical bond will be broken when the Chlorine component is detoxified, but the Chloramines will then release ammonia into the water. Additional binder may be required, as compared to source water with only chlorine or ammonia. Water treated for Chloramines should be tested for both residual Chlorine and Ammonia. This testing can be done as soon as the binder is mixed into the water, because the chemical bonds form on contact. An additive that detoxifies chlorine and ammonia at the same time would also bind ammonia produced by the Koi during transport [38]. 5.3 Slime Coat Enhancement


The need for slime coat enhancers when handling Koi has been discussed earlier in this paper. Even calm Koi that do not move much in the bag will lose some slime coat due to general banging around during transport. Koi that are stressed from poor water quality tend to be more agitated in the bag, and may lose more of their slime coat. Smaller Koi shipped together will bang against each other in the bag and lose slime coat. The slime coat released into the water degrades water quality by increasing organics and additionally stresses the Koi by clogging their gills, thus making it more difficult for them to extract oxygen from the water. Any chemical marketed as a slime-coat enhancer that works by irritating the Koi to produce more slime coat causes additional stress by coating the gills and making it even more difficult for the Koi to breath. Any pathogens in the transport water have free entry to attack the Koi in any area in which the slime coat is missing, and in warm water, harmful organisms may reproduce fast enough to damage the Koi even during relatively short transports. Any areas where a Koi’s slime coat is missing from netting, handling or rubbing against the bag will allow pathogens a portal of entry. Many antibiotics, sedatives and disinfectants have the derogatory effect of reducing slime coat, and damaging gill function. If a Koi’s slime coat is comprehensive and at healthy levels, it will help protect the Koi from pathogens. It is always a better strategy to prevent damage than to treat it. If slime coat is being replaced as soon as it is missing, the Koi will be safer from pathogens. 5.4 Treatment of Osmotic Stress As mentioned earlier, Koi must expend considerable energy keeping water out of their systems. The process of maintaining the water-to-salt balance in Koi is called osmoregulation. Even at rest, Koi must excrete excess water that enters primarily through their gills, and work at retaining salt to maintain osmotic balance [39]. Excess water in their bodies may overload their kidneys and cause death. Koi experiencing osmoregulation problems may appear to be bloated or even look like pinecones, with their scales sticking out, away from their bodies, and their eyes may bulge. While there may be many causes for osmoregulatory distress and failure, a simple way to reduce osmotic stress is to add salt (sodium chloride) to the water up to 0.3 % (see Table #4, Appendix). Salt must be completely dissolved before Koi are added. The salt reduces the gradient that the Koi’s body must fight to excrete water [39]. The addition of salt to the water decreases the amount of effort that is required to resist the uptake of excess water and maintain salt, thus increasing the amount of energy that the Koi has available to repair damage caused by stress. Other electrolytes including Calcium may also be used. (see Appendix on “Maintaining a Stable pH and Calcium” for a list of substances that may be used to increase Calcium) The salt will also work to stimulate slime coat production [50, 51, 52]. 5.5 Detoxification of Heavy Metals


Heavy metals often found in well water include copper, lead, zinc, chromium and mercury. While most municipal water is treated for heavy metals, copper or lead can find its way back into transport water if they are used in the plumbing. As long as water flows through the pipes regularly, the lead or copper does not build up in the water. If a pipe has not been used for a long time, copper may have leached into the adjacent water, and may reach toxic levels if that water is used to transport Koi. Always run water to waste for 10 minutes before filling transport bags or tanks. Heavy metals may be more toxic in a transport bag or tank because the severity of their detrimental effect depends on the ever changing water quality. 5.6 Disinfecting the Water Antibiotics such as bactericides, fungicides, and other disinfectants are sometimes used as bag additives to decrease the bacterial and fungal count in the bag [19, 43]. The authors of this paper would not recommend using disinfectants, because they are very hard to control and may have adverse effects. Many disinfectants that are effective enough to reduce parasites or bacteria are toxic to Koi, especially in the every changing chemistry of the shipping bag, or over a prolonged shipment period. Some substances interact with each other, and either nullify the good effects of each or become toxic in combination. The changing water pH can cause substances to become more or less effective or lose effect entirely, while others precipitate out of solution. Antibiotics used as bag additives have their own problems. According to Griffiths, “There are no real safe antibiotics; all antibiotics can kill if used inappropriately [40].” Antibiotics are targeted killers. Using a single antibiotic may have no effect, as it may not be effective against the existing strain of bacteria. Griffiths continues, “The mixing of antibiotics is not really desirable and should be discouraged wherever possible [40].” There are good bacteria that populate Koi as well, and antibiotics are indiscriminate killers of all susceptible bacteria – both good and bad. (for a further discussion of water borne antibiotics, see Appendix) 5.7 Buffering the Water Buffering is sometimes added to the water to keep the water stable while Koi are being transported. Changing water parameters are often linked to stress in Koi. The most common buffering agent is baking soda, or sodium bicarbonate [19]. Buffering transport water keeps the pH from dropping from the acidic affects of carbon dioxide and ammonia. A buffer will also increase total alkalinity as it stabilizes the water by keeping the pH constant. However, buffering is only recommended in transport water where all ammonia is being bound. If ammonia is not being bound, it is critical that the pH be allowed to drop so that the ammonia is less toxic. If ULTIMATE® is added to the transport water as recommended, it is performing the job of an ammonia binder. It is not necessary or desirable to use a buffering agent along with ULTIMATE®.


5.8 Increasing Oxygen Certain chemicals called oxidizers will react with water to release oxygen. If more oxygen is available, a Koi has to expend less energy to extract it from the water. Oxidizers may produce oxygen, but many also have been shown to damage the secondary lamellae of the Koi’s gills. In order for gills to work effectively, they contain millions of passages just wide enough for a single blood cell to pass through [8]. Blood cells are infused with oxygen in the narrow passages. An oxidizer can damage the structure of the passages, similar to the damage done by acids. While Koi gills have remarkable healing characteristics, once the damage has reached a certain level, it is irreversible. Hydrogen peroxide, a common oxidizer, was observed by Johnson [41] to reduce slime coat. Any oxidizer used for the period of transport may be doing much more harm than good. 5.9 Sedation Sedatives and anesthetics which relax the Koi and decrease their respiration rate have been used as bag additives [42]. While adding a sedative or anesthetic might initially sound like a good idea, some chemicals that cause sedation in Koi can have unpredictable effects for various reasons such as water chemistry, temperature, size of the Koi and the amount of fat in the Koi [1].

“Because drug dose and exposure time are often cumulative, it is difficult to maintain a uniform depth of anesthesia [during transport]. One reason for this is that levels of anesthesia may continue to accumulate in the brain and muscles even after blood levels have attained equilibrium [1].”

A high initial dose of sedative may stop a Koi from breathing. As Koi are sedated, the first stage is excitation, often accompanied by hyperactivity while the Koi fights to regain normalcy [1]. Excitation and increased activity will raise blood pressure and respiration. This could cause the Koi to inflict damage to themselves in the confines of the bag, or even to damage the bag itself. Another danger, as discussed under the Ammonia Stress section, is that the use of anesthetics will ultimately depress the metabolic level of the Koi, and they may not be able to expel enough ammonia to overcome ammonia autointoxication, which may be fatal. For all of these reasons, anesthetics or sedatives should never be used as bag additives. 5.10 Darkness Keeping the Koi in the dark during transport has been proven to reduce stress, and reduced stress means that Koi will be calmer and move less within the bag or tank. A calm Koi will not bang around as much, and will arrive with more slime coat intact [19]. 5.11 Additive Conclusions & Recommendations


In conclusion, an ideal bag additive would: Detoxify ammonia and chlorine Preferentially replace slime coat only where it is missing Continuously replace slime coat Detoxify Heavy Metals Buffer the water such that the water quality parameters are stable during transport Treat Osmotic stress An ideal bag additive would not: Risk damaging the Koi’s gills Decrease the Koi’s slime coat Excite the Koi or increase activity level Destroy beneficial bacteria Have effects that vary over time, temperature or water quality Depress the Koi’s metabolic rate and make it susceptible to ammonia autointoxication Any chemical that prevents one problem but causes another, it is not a good candidate as a transport additive. All goals can be achieved through the use of ULTIMATE® and salt as transport additives. The ideal shipping system would include: Good base water Ice packs, well insulated, adhered to the upper sides or top of the container ULTIMATE® sufficient to remove chlorine, chloramines, and ammonia produced by the Koi for twice the expected transport time Salt at .3% (see Table #4, Appendix), dissolved before Koi are added Pure oxygen to fill approx. 2/3 – 3/4 of the bag, leaving 5-10% bag space for expansion Koi kept in the dark during transport (If a transport tank is used instead of bags, an air pump with air stone can be used to provide oxygen. All other recommendations remain the same.) ULTIMATE®, along with the addition of salt up to 0.3% meets all the goals for transport additives, and there are no detrimental side effects. ULTIMATE® cannot be overdosed and cannot harm the Koi. The Koi should emerge from the transport bag or tank with a perfect layer of slime coat, and undamaged gills. If the Koi are transported in the dark, they should be calm and will have greatly reduced stress. Note: that the use of ULTIMATE® and salt may cloud water with certain chemistries, but clouding does not affect the effectiveness of either the ULTIMATE® or the salt [49].


6.0 EQUIPMENT AND SUPPLIES This section will deal with the supplies and techniques that have been found to most effectively transport Koi. 6.1 Equipment and Supply List: A Seine net to gather Koi into one corner of the pond A Koi pole net to guide Koi into a bowl A bowl of sufficient size to easily hold the largest Koi Koi Bags: at least 20” x 40”, 3 mil thick, clear plastic Bag Additives (ULTIMATE® and Salt) Strong Rubber Bands or Koi bag closure bands and applicator tool Ice packs, well wrapped in newspaper, bubble-wrap or other insulation Boxes lined on all sides with Styrofoam, or coolers Oxygen bottle with hose and a sterilized fill nozzle Emergency supplies 6.2 Seine net Horizontally, the seine net should be at least 4 feet larger than the maximum width of the pond, and in the vertical dimension, it should be 3 feet taller than the deepest part of the pond. A larger net is always preferable to one that is too small. The bottom should have sufficient weight such that the entire bottom edge of the net will remain on the pond bottom while the net is being slowly pulled through the water. The top of the net should be capable of extending at least 3 feet (higher is always better) above the water in order to turn back any jumping Koi. The mesh should be soft and the holes small enough to assure that the smallest Koi cannot become stuck. The mesh should be knotless to avoid injuring the Koi It is often easiest to roll each side of the net onto long poles that can be more easily guided along the pond sides, and thus extra width of the seine is recommended. The seine is gently placed in the water along the back edge of the pond, opposite to the area where the Koi will be removed, and allowed to fall to the bottom. A minimum of 2 people are required to seine most ponds. The assistant on each side of the pond then wraps their side of the net onto a pole, such that the floats on the top of the net extend above the water by 3 feet. The pole is held tight up against the wall on the side of the pond while it is slowly advanced from the back of the pond to the front. Wrapping the net on poles prevents gaps in the sides of the seine while the net is being moved, and assures that the bottom of the net stays on the bottom of the pond while allowing the top to extend above the water line to turn back jumping Koi. It is imperative that the seine is moved very slowly through the water. The slower that the seine is advanced, the easier it is to keep the edges of the net or the poles against the sides of the pond, and the easier it is for the weights to hold the net onto the bottom of the pond. Any gaps will quickly be exploited by the Koi, as Koi learn very quickly. If the Koi escape the net once, they will try again, and each time they are successful, the more determined they will become to escape


the next time. Koi that are calmly and efficiently collected with a seine don’t learn to jump or try to escape. If a Koi reacts to confinement by jumping and is turned back by the net, it will be less likely to jump again. Each time Koi are seined and people interact with them, there is an opportunity for the Koi to learn and to influence how the Koi behave. If the Koi have a low-stress experience, they will be even less stressed the next time the net is introduced. If they panic and jump and need to be re-seined several times, they will be in much worse physical condition from the stress, and it will be much harder to catch them the next time. The Koi should be collected into one corner of the pond where it will be easy to reach them with the pole net. Seine nets may even be used in irregular-shaped ponds, so long as the net can make several wraps around the poles being held on each side, and is still wide enough to stretch across the widest area of the pond. As the helpers advance to narrower areas, the extra net may be slowly wrapped onto the poles, and then unwrapped again where the pond gets wider. Extra care and time should be taken with irregular-shaped ponds. Ponds with islands or central features are not as suitable for seining. To seine ponds with islands, 2 seines are required. One is left in place to block the water between the island and one edge. The other net is dropped into the water near the stationary one, and is worked in a circle back to the stationary net to trap the Koi between the two nets. Seine nets may be adjusted for different bottom configurations by changing the amount of weight at the bottom of the net. Most seine nets can be rigged to ride over bottom drain covers without catching. Deeper and smoother ponds may require additional bottom weight. If the bottom is rocky or very irregular, weights may need to be removed. The heavier the bottom weights, the more quickly the seine net may be moved. Light seine nets require tremendous patience in order to move slowly enough to keep the net on the bottom at all times. 6.3 Pole Net The Pole Net should be a net specifically designed for handle Koi. The head of the net should be large and round and the netting should be quite shallow. A standard size Koi net would have a 25-inch diameter head, and the netting would be less than 10” deep. Most Koi nets have a 6-8 foot long handle. The length of the handle or pole should be compatible with the depth and size of the pond. Handles may be as long as 20 feet, although large nets with longer handles become heavy and hard to maneuver. The netting should be designed for Koi and should never be larger than a 3/8-inch mesh. The mesh should be made of polyester, and coated to resist barb penetration and scaling. The area on the frame where the mesh netting is attached should be designed in such a way as to prevent any possible injury to the Koi. Many ingenious methods have been developed to protect Koi from being injured, and there are many excellent net designs available to choose from. The net should be black to decrease its visibility to the Koi. The handle should be at least 2” in diameter and designed to reduce twisting. If the handle is made of wood, it should be oval in shape to resist twisting due to the resistance of the large head being moved through the water. If it is made of aluminum, it should


have a foam grip. Some nets have hollow metal handles which can fill with water and become very heavy, hindering the maneuverability of the net. Wooden handles float, and make the net lighter and easier to use. The head of the net should be larger than the largest Koi. The larger the head, the harder it will be to move through the water, and the slower it will be, due to water resistance and drag. Larger nets will try to twist as they are being moved. It is much easier to catch small Koi with larger nets. Smaller Koi can dart around very quickly and easily escape smaller nets. The key to catching Koi is to move the net very slowly under the Koi until the Koi is selected, and then use the net to guide the Koi towards the bowl. Position the net under the Koi’s head and as the Koi turns, continue to slowly follow the head (rather than the tail) while guiding the Koi to the surface. Koi catching is somewhat of an art, and good Koi wranglers learn to watch the Koi’s body language to indicate which way the Koi is going to turn. Following the head and moving slowly works well for most people handling larger Koi. The net should never touch the Koi, and keeping that goal in mind should help to keep all motion slow and calm. It is often desirable to use a net large enough for the net to then be used to cover the bowl while it is being moved, thus preventing injury to the Koi as a result of jumping. Koi should never be lifted out of the water with a net, regardless of the Koi’s size. All Koi depend on water to support their body weight, and the internal organs or larger Koi can be seriously damaged by being lifted them without proper support. 6.4 Koi Bowl Most Koi bowls are round and made of heavy blue plastic. The color blue has been found to provide the best contrast to Koi colors, and is the only color used at Koi shows. The blue color makes an excellent background for photographing Koi. The bowl should be larger in diameter than the length of the largest Koi. While the Koi is being guided towards the bowl by one person, another person should be holding the Koi bowl. The bowl is tipped vertically, so that 1/2 the bowl is in the water and 1/2 is out of the water. The open side of the bowl should face towards the Koi, with the bowl back facing the bowl handler. The biggest mistake made by ‘bowlers’ is that only the lip of the bowl is kept in the water, and the Koi must be forced up and over the lip to get into the bowl. The Koi then panics because the bowl does not have enough water, and no matter how carefully the Koi was guided to the bowl, it is now highly stressed. Keeping the bowl 1/2 in the water, and swimming the Koi into the bowl slowly, will greatly reduce stress. Once the Koi is in the bowl, the bowl is allowed to slowly tip to its normal appearance and float on the water. The bowl will be nearly full of water. More water may be added by tipping the edge of the bowl nearest the ‘bowler’ under water and allowing water to flow in to the desired level. Water may be removed by slowly raising the back of the bowl up in the air, and allowing excess water to run out of the front of the bowl towards the ‘bowler.’ This does not require strength - the total weight of the bowl should be supported by the water as it is floating. If the bowl is tilted slowly, the Koi will swim towards the flow into the back of the bowl and will not be poured out with the water. Some Koi have a tendency to jump from a bowl that has deep water. Generally, just enough water


should be used to cover the Koi’s dorsal fin. Once in the bowl, the Koi should be examined for health, and then bagged directly from the bowl. Most Koi farms will gently turn the Koi upside down and examine the belly for signs of disease. A small camera can be used instead, and will cause much less stress than the handling required to examine the Koi’s belly. If a Koi is agitated and resists handling for examination or being picking up, it often helps to make the Koi dizzy for a moment. This can be accomplished two different ways. First, one hand may be placed next to the Koi’s head and other hand near the tail to gently spin the Koi several times about the axis of its middle, until the Koi has lost its orientation and will stay in one place when it stops spinning. If the Koi bowl is in the water or on a very smooth surface, it may be preferable to simply use both hands to spin the bowl. This system has the advantage of not having to handle the Koi or accidentally remove slime coat, but may take a bit longer. In the moment of confusion after spinning, most Koi can usually be handled without a battle. If the bowled Koi passes its examination, it may be bagged. If there are any areas of redness or signs of disease or damage, the Koi should be moved to a quarantine tank and treated. Only healthy Koi should be shipped. 6.5 Koi Bags and Number-of-Koi-per-Bag Koi bags should be made of heavy, 3-mil clear plastic. Heavier plastic will make the bag hard to seal with the bands, and thinner plastic may stretch and tear when the bag is moved. Double bagging is recommended for Koi under 18”, and triple bagging is recommended for Koi over 18”. Small bags should not be used, even for small Koi. While there are many different sizes of bags available, most Koi farms would recommend using a bag that is 20-24” wide by 40-48” long. The general rule is that each Koi over 12” should go in its own bag with approximately 3-5 gallons of water. Koi should not be shipped together when the final health of the Koi is the only concern. Increased crowding in bags increases the stresses from the whole transport experience, and may contribute to Delayed Mortality Syndrome. For the purposes of transporting Koi to shows, all Koi should be packed individually. The innermost Koi bag should always be new. If it is desired to re-use bags, they should only be used as the 2nd and 3rd bags and never contact the Koi. Re-used bags may harbor disease, and it is very hard not to contaminate the new inner bag, and allow the possibility to pass on disease. Preferably, no Koi bag should ever be reused to insure that all bags are free from contamination. Bags should never be stored where they can be splashed by water containing Koi, again to avoid contamination. For all Koi bags, just enough water should be added to cover the gills (the dorsal fin may be slightly out of water), and the remainder of the bag should be filled with pure oxygen. Koi bags, after being banded, should always be at least 4” longer than the Koi, allowing 2” at the nose and 2” at the tail to buffer injuries. Larger bags and wooden crates will be required for Jumbo Koi. Some Koi Farms also suggest that the corners at the bottom of the bag be tucked under, as small Koi have been trapped and suffocated in these areas.


When moving Koi over 12”, both ends of the bag should be supported so that the spine of the Koi is kept straight during the entire process. Moving large Koi may require 2 or even 3 people to keep the Koi’s spine straight and to lift the larger amount of water. (information for this paragraph was given by the breeders and dealers interviewed in Table #3, Appendix) 6.6 Rubber bands Each Koi bag, whether it is used as the first, second or third bag should be sealed with three bands. To properly seal the bags, leave an 8-10” tail after the inside bag is filled with pure oxygen. This tail should be very tightly twisted to assure the best seal with the band. Either heavy duty rubber bands or special Koi bagging bands may be used. Typical rubber bands are 3.5-inch long and just under 1/4” wide. The width of the band is important – thinner bands will tend to cut the bags, and thicker bands are hard to get tight. If starting with a 3.5-inch rubber band, it is easier if the band is twisted in half so it is double-thick. It should still be possible to stretch it easily over the twisted neck. The band is stretched and twisted and placed over the neck again to make 4 thicknesses, and then as many more times as possible. This takes quite a bit of strength. This first band should be placed low on the neck away from the open end to prevent the neck from untwisting. The tightly-twisted tail of the bag should be doubled over and a second and third band should be used to very tightly secure the tail, thus providing a secondary seal. Each bag should be banded the same way, always starting with the inside bag and working towards the outside bag. A good adage to remember is that bags and rubber bands are cheap, while Koi are expensive. If hand strength is an issue, it may be easier to use a tool and Koi bagging bands. The tool looks like a pliers, but works in the opposite way, such that when the handles are squeezed together, the head opens. The head has 1 or 2 pins on each side. When the head is closed, a small and very stiff Koi bagging band is placed around the pins. The handles are squeezed together, and the band is expanded. The handles must be held against the force of the band while the band is slipped around the twisted neck of the Koi bag. These bands should always be used in a group of 3, just like regular rubber bands, with the second and third bands used around the doubled-back tail of the bag. These bands will need to be cut to be removed, which will likely result in pierced bags. Some people prefer to twist the neck of the bag, then double it back and use a single band only on the doubled neck. This system is not recommended. It is difficult to get a good seal against the doubled neck, and a disaster may occur if there is band failure. (information for this paragraph was given by the breeders and dealers interviewed in Table #3, Appendix) 6.7 Ice Packs Ice packs can be found in many sizes and shapes. It is always preferable to use one larger ice pack than many smaller ones. Ice melts faster if it has more exposed surface area, so larger ice packs last much longer. The ice pack is used as an air-conditioner in the Koi box, and should not be in direct contact with


the Koi bag. Ice packs should be wrapped in multiple layers of newspaper, bubble wrap or other insulation. Insulated ice packs prevent any sharp edges from puncturing the Koi bags, prevent injury to a jumping Koi, and slow the melting rate of the ice to make it last longer. Each standard 16 ounce reusable gel ice pack (usually 6 1/2” x 5 1/2” x 1”) will last between 4 and 8 hours, depending on the air and water temperatures. For short shipments under 6 hours, one ice pack should be sufficient. If the shipping time is longer than 6 hours or if the air temperature is above 75, multiple ice packs should be used. (information for this paragraph was given by the breeders and dealers interviewed in Table #3, Appendix) 6.8 Containers Koi should not be transported only in bags. The bag with the water and fish needs to be supported in a sturdy container. Unsupported bags roll around with the slightest movement, and even the forces encountered when making a corner can shift a heavy bag so violently that it will burst the bag or band. Koi can be badly injured in a sudden stop, and may severely damage fins during even a short transport without the stability provided by a container. It is important to have the correct size box for the Koi Bags that are being used. The Koi bags should lie on their sides on the bottom of the shipping container, with the banded necks at one end, and the bottom of the bags at the other end. Care should be taken to assure that there are no wrinkles in the bag. The bag should fill the entire bottom of the box so that it will not shift during transport. The box itself should have a minimum 300 lb. crush pressure rating, or at least be double walled. While Koi are not particularly heavy, water is. Water weighs over 8 lbs per gallon, and filled transport boxes usually weigh over 30 lbs. If that weight is dropped, it will exert more than enough pressure to burst a standard cardboard box. Most Koi shippers recommend using a heavy plastic liner on the bottom of the box that extends at least 6 inches up the sides. The liner will contain any water leaks and prevent the water from destroying the box. The box should be clearly marked to indicate the right side up, and should be labeled to indicate that live fish are enclosed. All sides of the box including the top should be insulated with Styrofoam to help maintain the air-conditioning effect from the ice pack(s). If ice packs are put on the bottom of the box, a heavy piece of Styrofoam should be placed over the ice packs to support the Koi bags. If the Styrofoam tips and is unstable on top of the ice packs, the ice packs can be secured to the sides or top of the box with several pieces of duct tape. Coolers are used instead of boxes for the transport of valuable Koi. The coolers are much more crush resistant, hold water, and will keep a more consistent inside temperature in warmer conditions. Some have built-in handles and easy-to-close lids that make coolers easier to work with. Ice packs should be well secured to the sides or top of the cooler with duct tape. Empty space in both boxes and coolers should be filled with bubble wrap so that bags cannot shift during the journey.


Koi containers should always be placed perpendicular to the direction of travel. This is especially important with large Koi that cannot turn around in their bags. If the container is parallel to the direction of travel and makes a sudden stop, the Koi may badly injure its nose or tail. Shipping Koi sideways is more comfortable for the Koi and prevents injury. It is never advisable to put Koi containers in car trunks. A trunk may be subject to extreme temperatures – both hot and cold. Bubble wrap should be used to line all sides around bags that contain heavy or large Koi. The bubble wrap will greatly cushion any sudden movements and prevent injury to the Koi. This is more important in a hard sided cooler, but should also be practiced when using a box for heavy or large Koi. (information for this paragraph was given by the breeders and dealers interviewed in Table #3, Appendix; also see [19, 38, 43]) 6.9 Oxygen The amount of pure oxygen in the shipping bag is more important to Koi survival then the amount of water. The purpose of the water is to provide a medium to get oxygen to the Koi’s gills. Water holds very little oxygen. Increased oxygen in the water makes it easier for the Koi to breathe during their journey. The best way to make sure that there is enough oxygen is to replace the air in the bag with pure oxygen. Air contains only 20% oxygen. Filling a Koi bag with pure oxygen provides 80% more oxygen that can be dissolved into the water for the Koi to breathe, thus allowing the Koi to survive for a very long time in a very small amount of water. A larger Koi Bag simply can hold a greater amount of oxygen. In large shipping bags filled with pure oxygen, medium sized Koi have been known to survive over 72 hours when the properly prepared shipping boxes were delayed. Cold water can hold more oxygen (see Table # 2, Appendix) than warm water, and is another reason to ship Koi in cooler water, and add pure oxygen to the bag. Koi owners should keep a tank of pure oxygen for emergencies and transport. A cylindrical oxygen tank can be obtained from any welding supply store, or smaller tanks may be purchased from any home improvement store. Filled tanks from welding stores are usually rented. If a large tank is chosen, the company will come out and refill the tank at the user’s request. Small empty tanks are usually exchanged for filled ones. When transporting oxygen cylinders, it is imperative that they be snugly restrained and transported on their sides so that they cannot fall and the regulator cannot become dislodged. Damaged oxygen tanks may be extremely hazardous, and all precautions must be taken each time a tank is moved. All tanks should have a regulator and gauge that shows the pressure of the oxygen and the amount of oxygen remaining in the tank. Make sure the filler hose is long enough to easily fill Koi bags without the cylinder being in the way. The hose should have a special air-filling valve at the end. Once the main tank valve is turned on, the air is supplied by gently squeezing the filler-valve at the end of the hose. Koi Bags need to be filled slowly, or the agitation and noise will scare the Koi. Always remember to turn off the main tank valve when filling is complete, or the tank may be empty the next time it is needed. Filler-valves on hoses are not meant to hold the pressure of the tank for long periods. Remember to sterilize any portion of the filler valve that comes into contact with Koi water. It is recommended to use a short length of plastic tubing attached to


the filler valve, which should be changed or disinfected between shipments to diminish the risk of spreading pathogens. Oxygen is only added to the inner Koi bag; the one actually containing the water and Koi. The bag should be purged of air and filled with pure oxygen. To accomplish this, hold the neck of the bag loosely in one hand. Use the other hand to very gently press the surface of the bag onto the water, allowing all the air to escape from the neck of the bag. Then grip the neck of the bag tightly to prevent any room air from entering the bag. Carefully insert the plastic tube attached to the air-filling nozzle inside the neck of the bag, and clamp the bag tightly around the nozzle to prevent room air from entering the bag. Slowly squeeze the air-filling valve and expand the bag to nearly full. Oxygen should be added along the side of the bag slowly enough to not distress the Koi. When the bag is nearly full, remove the air-filling valve and quickly twist the neck of the bag to prevent the pure oxygen from escaping. Place the bands on the inner bag according to the above instructions, and then band the outer bag(s). If the Koi are to be transported by air, it is imperative to leave some space in the bag, and not completely fill the bag with oxygen. The air inside the bag will expand if the transport vehicle travels to higher ground and the outside air pressure decreases. An airline baggage compartment is kept pressurized – but only to a pressure corresponding with 8,000 feet. If the Koi are bagged at sea-level and then the container is placed in a baggage compartment, the reduced pressure of the compartment air (similar to 8000 feet) will allow the higher pressure air inside the bag to expand about 5%. If sufficient room is not left inside the bag for this expansion, the bag will burst, and the water will leak out, killing the Koi. A Koi bag prepared for air shipment should have at least a 10% space for expansion. If Koi are transported by ground, 5% expansion room should be left if the terrain changes by less than 5000 feet, and a 10% expansion space should be left if the terrain will change by more than 5000 feet. The only time it is acceptable to completely fill a bag with air such that the bag feels hard, is when driving Koi across town. A good rule of thumb is that a bag filled with oxygen should always have 10% expansion space. 6.10 Emergency Kit It is always advisable to carry an emergency kit when transporting Koi. That kit should include a 5 gallon container of shipping water, a small bottle of ULTIMATE®, extra Koi bags, extra bands, and a small oxygen cylinder with filler valve. While the bags and bands will fold into a tiny packet, traveling with an oxygen cylinder can be a risk some owners prefer not to take. Some owners choose instead to carry a cooler and a small battery operated air pump and air stone. Small battery operated air pumps with air tubing and an air stone are sold in the fishing departments of large discount stores, and are used for bait buckets. 2 D-cell batteries will run one of these small air pumps and produce enough air to keep a Koi alive for several hours. If a bag were to burst or leak, the emergency kit should contain all the equipment needed to keep the Koi alive until the destination is reached.


If a Koi is found alive, but has been out of water for some time, the Koi can sometimes be saved by giving an IM injection of equal parts Dexamethasone and liquid Vitamin C (recommended by Dr. Sandra Yosha, DVM, PhD – see Biographies, Appendix). The syringe, drugs and proper dosing should be obtained from a local Koi veterinarian. Note that Dexamethasone is a steroid that can quickly reduce the swelling to tissues caused by exposure to air [43], but can have detrimental effects on the Koi’s immune system. Thus, it may be paired with Vitamin C to minimize the detrimental effects. Once the shot is given, the Koi must be revived. To revive the Koi, water must be moved in the mouth and out the gills, as would occur in a normal swimming motion. This can be accomplished by holding the Koi upright and slowly moving the Koi forwards through the water until it tries to swim on its own. (Never swim a Koi backwards through the water, as it may damage the gills.) Alternately, a tiny fountain-size water pump can be used. A small piece of soft plastic tubing is attached to the pump outlet and the end is put into the Koi’s mouth. While it may take Koi as long as 45 minutes to revive, Koi have been known to make full recoveries from being dry for up to an hour. 6.11 An Alternative – Live Haul Tanks Most Koi farms use live haul tanks to transport Koi between locations. These tanks are often custom made to fit onto farm trucks, but there are also reasonably-priced models that are available in moderate sizes for the consumer. Consumer models feature a frame that is constructed of sturdy PVC pipe, which supports a tank made of heavily-treated plastic fabric - identical to that used for show tanks. Ideally, a transport tank would have soft sides to minimize any impact damage to the Koi. A spare truck battery along with a power inverter may be used to run any efficient air pump and air stone. The truck battery and inverter may also be used for power emergencies around the home and pond. Be sure that the air pump is located far from the vehicle exhaust such that the intake cannot pick up carbon monoxide which is very soluble in water and very deadly to Koi. When choosing the water to be used in transport, the considerations are the same whether the water is used for bags or live-haul tanks, and shipping tank water should also be amended with ULTIMATE® and salt before the Koi are added. Most Koi farm managers use transport tanks, and are convinced that the Koi are less stressed when transported by tank. It is certainly far less labor-intensive to move Koi into-and-out-of a transport tank than to properly bag and box the Koi. Tank weight is the primary consideration, as not all vehicles have the facilities to carry or hold the weight securely. These tanks are quite unwieldy, and weigh over 400 lbs with only 50 gallons of water. Note that water weighs 8.3 pounds per gallon, and a fully loaded transport tank may exceed the weight limits of many vehicles and trailers. Larger transport tanks often contain baffles. While these may restrain water from sloshing and creating dangerous driving conditions, they may also injure Koi [44]. While metal tanks are less likely to be damaged or broken, the welds in a tank may break if the truck-bed warps. These issues are usually handled by the use of several smaller tanks, placed side by side.


Tanks with large doors or lids make loading and unloading much easier and reduce the rough handling sometimes associated with net dipping, especially in tight quarters. Extreme care must be taken to reduce stress while getting Koi into and out of transport tanks. All transport tanks should have safety features including overflow-drains and air vents that allow the dispersal of carbon dioxide, a by-product of Koi waste. All transport tanks must have an aeration system to ensure the addition of adequate dissolved oxygen into the water, and precautions should be made to have a back-up power source for the aeration. An overcrowded transport tank without oxygen quickly becomes deadly to Koi.

7.0 SUMMARY – 50 Things You Should Know About Transporting Koi 7.1 Stress and Koi Stress caused by handling and transporting Koi can be considerable. Multiple stressors have an additive effect. Each stressor may reduce the Koi’s ability to tolerate subsequent or additional stressors. Koi may succumb to Delayed Mortality Syndrome within a 2-3 week period after the acute stress of shipping. Koi may not fully recover from all the stresses involved for shipping for several months, and should thus not be shipped any more frequently than every 3 months. Koi are very adaptable and may be conditioned to adapt to normally stressful events, such as netting and bowling. Stress in Koi may be tested and quantified by swiping a Hemastix® Reagent Strip in the slime coat, and reading the color-change from the insert. Disruptions in the slime coat are thought to be one of the most important factors for why Koi succumb to pathogens after stress. ULTIMATE® does not coat the gills of Koi, but does appear to reduce the time it takes Koi to recover from stress. ULTIMATE® cannot be over-dosed. 7.2 Preparation to Transport Prior to shipment, Koi should be fasted for a week; in a holding tank, with a buddy.


Treat the holding tank with sufficient ULTIMATE® to neutralize any chlorine/chloramines plus all ammonia produced by the Koi. Maintain stable temperature, pH and total alkalinity in the holding tank, and add calcium to reach 80 ppm if it does not already exist in the water. Do not crowd Koi in a holding tank – allow 100 gallons per adult Koi. To minimize stress caused by different water quality parameters, test and adjust the critical parameters for all the water before the move: - Test the source water for Ammonia, Nitrite, pH, Alkalinity and Temperature. - Test the transport water (if not using the source water) and the destination water for pH, Alkalinity and Temperature. The goal should be to try to attain pH differences of less than 0.2 pH points and temperature differences of less than 5 Degrees F. Where adjustments need to be made to narrow the variation, it is preferable to raise the lower parameter than to lower the higher one. Ammonia and Nitrite should always be zero. Koi fasted for a week prior to shipping will produce less ammonia and require less oxygen in the bag. Never transport Koi with gill damage or any existing sign of injury or disease. Always carry emergency supplies while shipping Koi. The emergency kit should contain a 5-gallon container of shipping water, a small bottle of ULTIMATE®, extra Koi bags, extra rubber bands, and a small oxygen cylinder with filler valve. 7.3 Netting and Bowling Koi Take your time while netting Koi in order to keep stress at a minimum. Move slowly and steadily the entire time you are working with the Koi so that the ‘fight or flight’ response is not stimulated. Before bowling any Koi, have all the handling equipment ready and the transport equipment prepared. Use a seine net to gather Koi into one corner of the pond, where the Koi pole net can easily reach them. Use a Koi pole net to slowly guide the Koi into a Koi bowl, preferably being controlled by another person. Bag Koi directly from the bowl when possible. If the source water is to be used as transport water, the bag may be dipped into the bowl to collect the Koi. If the bag water is to be different than the source water, move the Koi by hand.


7.4 Shipping Bags Always use a new bag to hold the Koi to minimize the risk of spreading pathogens. Previously used bags may be employed as the 2nd or 3rd bags, if desired. For smaller Koi and shorter trips, use 2 shipping bags. For larger Koi or longer trips, 3 bags should be used. Always run water to waste for 10 minutes to flush any build-up of heavy metals from the pipes before using it as shipping water. Use a double-dose (per label instructions) of ULTIMATE® in the shipping water. Use a 4x dose of ULTIMATE® if chlorine or chloramines will need to be neutralized in the shipping water. Salt the shipping water to .3%. Add the salt and ULTIMATE® to the bag water and dissolve salt completely before Koi are added. Do not use disinfectants, antibiotics, oxidizers, anesthetics or sedatives as bag additives in shipping water. In the shipping bag, use just enough treated water to cover the Koi’s gills. Fill the remainder of the shipping bag with pure O2, leaving a 10% expansion space. Fill slowly along the side of the bag to avoid scaring the Koi. Bag each Koi separately in a 20” x 40” bag, or larger bag for larger Koi. There should be a minimum of 2 extra inches of space for the Koi at each end of the bag. Bubble-wrap should be used to line all sides around bags that contain heavy or large Koi to reduce the possibility of injury from a sudden stop during transport. Bubble wrap should be used to prevent any Koi bag from shifting during transport. Use 1 rubber band or Koi bagging band on the twisted neck of the bag, then double the twisted bag neck over, and use 2 more bands to secure it. Band each shipping bag separately. Once Koi are bagged, they should be kept dark and must be kept in the shade. Transport Koi in the dark.


7.5 Shipping Containers Either insulate all sides of Koi boxes with Styrofoam, or use a cooler to ship Koi. If cardboard boxes are used to ship Koi, the box should be double-walled or have a minimum 300 lb. crush rating. Use approximately one 16 ounce reusable gel ice pack (usually 6” x 5 1/2” x 1”) per 4 hours of shipping time to keep Koi cool during shipment. Ice packs should be insulated with newspaper or bubble-wrap and secured with duct tape to the upper part of the container to provide air-conditioning. Ice should never be in contact with the part of the bag holding the water. Place shipping containers perpendicular to the direction of travel. Never ship Koi in a car trunk due to extreme temperatures. If live-haul or transport tanks can be safely secured and carried by a vehicle, they may be used as an alternative to boxes and bags. Tanks must have an aeration system with backup power source, and should have overflow drains and air vents. Extreme care must be taken to reduce stress while getting Koi into and out of these tanks. 7.6 Un-bagging Koi When Koi arrive at their destination, place the bags into the quarantine tank and allow the bags to float in the shade for 15-30 minutes to equalize temperatures. Gently cut open the bags, and immediately lift out the Koi by hand such that none of the bag water gets into the quarantine or destination tank. Dispose of the bag water safely. 7.7 Recovery and Quarantine after Transport Use ULTIMATE® in the quarantine tank in sufficient quantity to: Neutralize all ammonia produced by the Koi Neutralize existing chlorine/chloramines. Maintain stable temperature, pH and total alkalinity in the quarantine tank, and add calcium to reach 80 ppm if it does not already exist in the water.


If the Koi are highly stressed or injured by the shipping, add .3% salt to the quarantine tank to reduce osmotic load and allow the Koi to recruit more resources for healing. Dissolve salt completely before Koi are added. Do not crowd Koi in a quarantine tank – allow 100 gallons per adult Koi. Quarantine the Koi for 30 days at 75 degrees. Offer no more than a few food pellets twice a day for each Koi in the Quarantine tank.


References [1] S. D. Coyle, R. M. Durborow and J. H. Tidwell, “Anesthetics in aquaculture,”

SRAC – Southern Regional Aquaculture Center, Publication Number 3900, November 2004.

[2] B. A. Barton, “Stress in fishes: a diversity of responses with particular

reference to changes in circulating corticosteroids,” University of South Dakota Department of Biology and Missouri River Instititute, Vermillion, South Dakota, 2002.

[3] T. D. Bartelme, “Stress in fish, part 1: what is stress in fish?” [Online].

Available: http://www.advancedaquarist.com/issues/july2004/short.htm, 2001.

[4] T. D. Bartelme, ”Stress in fish, part 2: why should you care about stress in

fish?” [Online] Available: www.advancedaquarist.com/issues/sept2004/fish2.htm, 2003.

[5] S. Bonga, “The stress response in fish,” University of Nijmegen Department

of Animal Physiology, Nijmengen, The Netherlands, 1997. [6] M. K. Stoskopf, “Environmental requirements and diseases of carp, Koi, and

goldfish,” in Fish Medicine, M. K. Stoskopf, Philadelphia, PA, W. B. Saunders Company, 1993, pp. 454-461.

[7] E. J. Noga, Fish Disease Diagnosis and Treatment, Iowa State Press

Edition, Iowa, Iowa State Press - A Blackwell Publishing Company, 2000, pp. 62-66.

[8] E. J. Noga, [7] pp. 22-25. [9] T. Sink, R. Strange, H. Kattesh and A. Mathew, “Linking stress to increased

mortality of channel catfish at varying concentrations of Edwardsiella ictaluri,” University of Tennessee Department of Animal Sciences, Knoxville, Tennessee, 2006.

[10] Z. Svobodova and B. Vykusova, “Diagnostics prevention and therapy of fish

diseases and intoxications,” Research Institute of Fish Culture and Hydrobiology, Vodnany, Czechoslovakia, 1991.

[11] Z. Svobodova, R. Lloyd and J. Vykusova, “Water quality and fish health,”

Food and Agriculture Organization of the United Nations, Rome, Italy, [Online]. Available: http://www.fao.org/docrep/009/t1623e/t1623e00.htm, 2003.


http://www.advancedaquarist.com/issues/july2004/short.htm

http://www.advancedaquarist.com/issues/sept2004/fish2.htm

http://www.fao.org/docrep/009/t1623e/t1623e00.htm

[12] R. Passovoy, “Snag ‘em, bag ‘em, and drag ‘em,” Midwest Pond and Koi Society, [Online] Available: http://www.mpks.org/articles/BobPassovoy/drag_em.html, n.d.

[13] E. J. Noga, [7] pp. 238-239. [14] Z. Svobodova, P. Kalab, L. Dusek, B, Vykusova, J Kolarova and D.

Jaaouskova, “The effect of handling and transport on the concentration of glucose and cortisol in blood plasma of common carp,” Research Institute of Fish Culture and Hydrobiology, Czech Republic, 1999.

[15] G. K. Iwama, L. Afonso and M. M. Vijayan, “Stress in fish,” presented at

AquaNet Workshop on Fish Welfare, B.C., Canada, 2004. [16] H. Selye, “A syndrome produced by diverse nocuous agents,” McGill

University Department of Biochemistry, Montreal, Canada, 1936. [17] E. L. Johnson, Koi Health and Disease, 2nd Edition, Athens, Georgia,

Reade Printers, 2006, p. 88. [18] A. C. Smith and F. Ramos, “Occult haemoglobin in fish skin mucus as an

early indicator of stress,” Journal of Fish Biology, volume 9, issue 6, pp. 537-541, 1976.

[19] G. Ewy, “Koi transportation, how to move Koi from here to there,”

Associated Koi Clubs of America, [Online]. Available: http://www.akca.org/library/transp.htm, n.d.

[20] N. Saint-Erne, Advanced Koi Care, Las Vegas, Nevada, Metagraphics,

2003, pp. 89-94. [21] B. G. Hecker, “Carp, Koi and goldfish taxonomy and natural history,” in Fish

Medicine, M. K. Stoskopf, Philadelphia, PA, , W. B. Saunders Company, 1993, pp. 442-447.

[22] S. Cover, “Koi physiology,” for the KHA Program, [Online] Available:

http://www.akca.org/kht/physiol.htm, 2005. [23] M. Smutna, L. Vorlova and Z. Svobodova, “Pathobiochemistry of ammonia

in the internal environment of fish,” [review], Research Institute of Fish Culture and Hydrobiology, Chech Republic, 2002.

[24] G. A. Wedemeyer, Physiology of fish in intensive culture systems, New

York, New York, Chapman and Hall, 1996, pp. 15 &16. [25] A. M. Phillips and D. R. Brockway, “Effect of starvation, water temperature

and sodium amytal on the metabolic rate of brook trout,” The Progressive Fish-Culturist, volume 16, issue 2, pp. 65-68, October1954.


http://www.mpks.org/articles/BobPassovoy/drag_em.html

http://www.akca.org/library/transp.htm

http://www.akca.org/kht/physiol.htm

[26] B. N. Bolon, “Special anatomy and physiology of goldfish, carp and Koi,” in Fish Medicine, M. K. Stoskopf, Philadelphia, PA, W. B. Saunders Company, 1993, pp. 447-450.

[27] E. J. Noga, [7] pp. 224-225. [28] S. Cover, “Koi Herpesvirus (KHV),” 2006, [online] Available:

http://www.akcaprojectkhv.org/index.php?option=com_content&view=frontpage&Itemid=1, under the pull-down “KHV Resources,” “Koi Herpesvirus.”

[29] D. Griffiths, Step by Step Advanced Koi Diagnosis and Treatment, UK, Des

Harris Associates, 2006, p. 6. [30] S. Yosha, “Update on Koi herpes virus (KHV) for the Koi hobbyist,” insert to

Koi USA, March/April 2003, [online] Available: http://www.akcaprojectkhv.org/index.php?option=com_content&view=article&id=46-Koi-herpes-virus-khv-a-threatening-disease-&catid=4-khv-resources&Itemid=20.

[31] K. H. Hartman, R. Yanong, B. D. Petty, R. Francis-Floyd and A. C. Riggs,

“Koi herpes virus (KHV) disease,” University of Florida IFAS - The Institute of Food and Agricultural Sciences, [Online] Available: http://edis.ifas.ufl.edu/vm113, 2004.

[32] C. Neaves, “Nutrition” for the KHA Program, [Online] Available:

http://www.akca.org/kht/nutrit.pdf, 2002. [33] A. Linden, M. Verhoye and G. E. Nilsson, “Does anoxia induce cell swelling

in carp brains? In vivo MRI measurements in Crucian Carp and Common Carp, The Journal of Neurophysiology, volume 85, number 1, pp. 125-133, January 2001, [online] Available: http://jn.physiology.org/cgi/content/full/85/1/125.

[34] E. L. Johnson, [17] p. 28. [35] E. L. Johnson, [17] pp. 86-87. [36] E. L. Johnson [17] p. 194. [37] M. Lammens, The Koi Doctor, Your guide to keeping healthy Koi, A-

Publishing, 2004, p. 20. [38] T. C. Crosby, J. E. Hill, C. V. Martinez, C. A. Watson and R.E. Yanong, “On-

farm transport of ornamental fish,” UF/IFAS - University of Florida Institute of Food and Agricultural Sciences, Florida, n.d.

[39] D. Griffiths, [29] pp. 127-129. [40] D. Griffiths, [29] p. 107.


http://www.akcaprojectkhv.org/index.php?option=com_content&view=frontpage&Itemid=1

http://www.akcaprojectkhv.org/index.php?option=com_content&view=frontpage&Itemid=1

http://www.akcaprojectkhv.org/index.php?option=com_content&view=article&id=46-koi-herpes-virus-khv-a-threatening-disease-&catid=4-khv-resources&Itemid=20



http://edis.ifas.ufl.edu/vm113

http://www.akca.org/kht/nutrit.pdf

http://jn.physiology.org/cgi/content/full/85/1/125

[41] E. L. Johnson [17] p. 135. [42] T. C. Crosby, J. E. Hill, C. V. Martinez, C. A. Watson and R.E. Yanong,

“Preparation of ornamental fish for shipping,” UF/IFAS - University of Florida Institute of Food and Agricultural Sciences, Florida, n.d.

[43] E. L. Johnson [17] p. 119. [44] G. L. Jensen, “Transportation of warmwater fish; equipment and guildlines,”

SRAC – Southern Regional Aquaculture Center, Publication Number 390, 1990.

[45] L. J. Stewart, “Nutrition of Koi, carp and goldfish,” in Fish Medicine, M. K.

Stoskopf, Philadelphia, PA, , W. B. Saunders Company, 1993, pp. 461-470. [46] E. L. Johnson [17] p. 80. [47] K.M. Treves-Brown, Applied Fish Pharmacology, The Netherlands, Kluwer

Academic Publishers, 2000. [48] E. J. Noga, [7] p. 72. [49] J.F. Kuhns and T. Pyle, AquaScience Research Group, Inc., North Kansas

City, MO, personal communications, August 2007 - September 2008. [50] E. L. Johnson [17] p. 129. [51] R.T. Di Giulio and D.E. Hinton, The Toxicology of Fishes, Boca Raton,

Florida, Taylor & Francis, 2008, p. 76. [52] C.S. Tucker and J.A. Hargreaves, Biology and Culture of Channel Catfish,

Amsterdam, The Netherlands, Elsevier 2004, p. 353. [53] E. J. Noga, [7] p. 295.


Glossary Ammonia autointoxication A rise in the ammonia levels in the blood caused by the reduced ability to eliminate ammonia. It is always recommended to fast Koi prior to transport, because when there is less ammonia in the bloodstream from the processing of food, there is less opportunity for autointoxication to occur. Various extremes in water parameters, toxins, damage to gill structure caused by pathogens and stress may all reduce respiration which reduces the ammonia transfer from the gills. The elimination rate of ammonia is a function of the health of the gill structure and the metabolic levels of the Koi. When Koi are then cooled in preparation to or during shipping, the ammonia builds in the bloodstream because the cooler temperatures have reduced respiration and therefore reduced ammonia transfer from the gills. This is why autointoxication may occur during transport. Chloramine When chloramine treated water is treated with a standard de-chlorinating agent, such as Sodium Thiosulfate, it releases toxic ammonia into the water. The ammonia must either be removed by the filtration system, or through the use of an ammonia binder. NH2Cl is commonly used in low concentrations as a disinfectant in municipal water systems as an alternative to chlorination. This application is increasing. Chlorine (sometimes referred to as Free Chlorine) is being displaced by chloramine, which is much more stable and does not dissipate from the water before it reaches consumers. NH2Cl also exhibits less of a tendency to convert organic materials into chlorocarbons such as chloroform and carbon tetrachloride. Such compounds have been identified as carcinogens and in 1979 the U.S. EPA began regulating their levels in U.S. drinking water. Furthermore, water treated with chloramine lacks the distinct chlorine odor of the gaseous treatment and so has improved taste. Cold Blooded The term is now outdated in scientific contexts. Cold-blooded creatures were, initially, presumed to be incapable of maintaining their body temperatures at all. Cold-blooded animals are now called either poikilotherms or ectotherms. Koi are properly classified as poikilotherms. (see Poikilotherm) Disease A condition of ill health or malfunctioning, especially one having particular symptoms. Hyponatremia Low blood salts, or an electrolyte disturbance (disturbance of the salts in the blood) in which the sodium concentration is too low caused by excess water accumulating at a higher rate than it can be excreted.


http://en.wikipedia.org/wiki/Concentration

http://en.wikipedia.org/wiki/Disinfectant

http://en.wikipedia.org/wiki/Municipal_water_system

http://en.wikipedia.org/wiki/Chlorination

http://en.wikipedia.org/wiki/Chlorine

http://en.wikipedia.org/wiki/Water

http://en.wikipedia.org/wiki/Chlorocarbon

http://en.wikipedia.org/wiki/Chloroform

http://en.wikipedia.org/wiki/Carbon_tetrachloride

http://en.wikipedia.org/wiki/Carcinogen

http://en.wikipedia.org/wiki/EPA

http://en.wikipedia.org/wiki/Electrolyte_disturbance

http://en.wikipedia.org/wiki/Sodium

Ectotherm, Ectothermic A pattern of temperature regulation where the animal’s body temperature depends mainly on using behavior to control the exchange of heat with the environment. A term which signifies that their heat (therm) comes from outside (ecto) of them. Examples of ectotherms are alligators and lizards that make use of the sun, shade and water as needed to adjust their body temperature. All ectotherms are also poikilotherms. Koi are poikilotherms, but not ectotherms. (see Poikilotherm) Hemoglobin Hemoglobin (also spelled haemoglobin and abbreviated Hb or Hgb) is the iron-containing oxygen-transport metalloprotein in the red blood cells of vertebrates. Hemoglobin transports oxygen from the lungs or gills to the rest of the body, such as to the muscles, where it releases the oxygen for cell use. Hyponatremia Low blood salts, or an electrolyte disturbance (disturbance of the salts in the blood) in which the sodium concentration is too low caused by excess water accumulating at a higher rate than it can be excreted. Moving Koi Moving Koi within a premises. Unless the health of the Koi is in peril, the authors of this paper would not recommend moving Koi more often than every 3 days, in order for the cortisol levels to return to normal. (see Shipping Koi) Osmoregulation, Osmoregulate Keeping the body's fluids from becoming too dilute or too concentrated. Osmoregulation is the active regulation of the osmotic pressure of bodily fluids to maintain the homeostasis of the body's water content; that is it keeps the body's fluids from becoming too dilute or too concentrated. Pathogen Infectious agent, or more commonly germ, is a biological agent that causes disease or illness to its host. The term pathogen is derived from the Greek "that which produces suffering." The body contains many natural defenses against some of the common pathogens in the form of the immune system and by some "helpful" bacteria present in the body's normal flora. However, if the immune system is damaged, or if the "good" bacteria are damaged in any way (such as the use of antibiotics being taken to kill other pathogens), pathogenic bacteria (that were being held at bay) can then proliferate and cause harm to the host. Poikilotherm, Poikilothermic An animal without effective autonomic temperature regulation such that their body temperature varies widely as a function of ambient temperature. Koi are poikilotherms. The outdated term “cold-blooded” does not describe how the animal accomplishes thermal regulation. If the animal’s behavior causes them to make use of sun, shade, and water to help regulate their internal temperature, for example, then they would ectothermic-poikilotherms. (see Ectotherm)


http://en.wikipedia.org/wiki/American_and_British_English_spelling_differences#Simplification_of_ae_.28.C3.A6.29_and_oe_.28.C5.93.29

http://en.wikipedia.org/wiki/Iron

http://en.wikipedia.org/wiki/Oxygen

http://en.wikipedia.org/wiki/Metalloprotein

http://en.wikipedia.org/wiki/Red_blood_cell

http://en.wikipedia.org/wiki/Vertebrate

http://en.wikipedia.org/wiki/Lung

http://en.wikipedia.org/wiki/Gill

http://en.wikipedia.org/wiki/Muscle

http://en.wikipedia.org/wiki/Electrolyte_disturbance

http://en.wikipedia.org/wiki/Sodium

http://en.wikipedia.org/wiki/Osmotic_pressure

http://en.wikipedia.org/wiki/Homeostasis

http://en.wikipedia.org/wiki/Water

http://en.wikipedia.org/wiki/Germ

http://en.wikipedia.org/wiki/Biological_agent

http://en.wikipedia.org/wiki/Disease

http://en.wikipedia.org/wiki/Illness

http://en.wikipedia.org/wiki/Host_%28biology%29

http://en.wikipedia.org/wiki/Greek_language

http://en.wikipedia.org/wiki/Immune_system

http://en.wikipedia.org/wiki/Bacteria

http://en.wikipedia.org/wiki/Human_flora

http://en.wikipedia.org/wiki/Antibiotic

http://en.wikipedia.org/wiki/Bacteria

Shipping Koi Any time the Koi are bagged and boxed. The authors of this paper do not recommend shipping Koi more often than every 3 months. (see Moving Koi) Stress A condition in which the dynamic equilibrium of animal organisms, called homeostasis, is threatened or disturbed as a result of the actions of intrinsic stimuli, commonly defined as stressors [2]. Stress may be quantified as the amount of disturbance of homeostasis. Stress should be considered cumulative - becoming greater by stages or additions. Stress is typically not a single phenomenon. Stress Response Koi’s physiological response to stress or stressors. Koi display a wide variation in their physiological responses to stress. Catecholamine changes (primarily adrenalin and noradrenalin) are typically an immediate response to stress and cause increases in cardiac output, blood sugar, respiration, oxygen uptake, and blood flow to the gills. Corticosteroid changes (primarily cortisol) are typically a response to chronic stress and affect the Koi’s osmotic balance and metabolism. The stress response in fish includes a marked increase in the oxygen uptake rate of the gills. Stress that is severe or chronic can cause harmful physical changes in the gill structure [5]. Stress also increases the permeability of the surface epithelium, including the gills, to water and ions, and thus induces systemic osmoregulation disturbances [9]. Stressors affect the branchial structure of the gills and result in osmoregulation imbalance. Stressors Any chemical, physical or perceived threat that triggers a stress response. Stressors can be multiple or continuous and vary in intensity. The more intense the stressor, the more likely it is that the fish may finally be in a distressed state. Torpor Torpor is an endogenous suppression and activation of metabolic processes creating a dormant state where dormancy is interrupted with periodic arousals, indicating that torpor is not temperature driven. During the periods of arousal, fish will eat flora and fauna living on the bottom of the pond. In cool water temperatures, a Koi’s metabolism slows down and they appear to go dormant. That condition is correctly called torpor. Koi do not hibernate, they go into torpor.


A P P E N D I X


Table #1 Relative Ammonia Toxicity as a Function of Temperature and pH

The table above shows relative ammonia toxicity as a function of temperature and pH. Because the un-ionized form of ammonia (NH3) is the toxic form, and because this form is more prevalent at higher pH and temperatures, ammonia becomes dramatically more toxic with increasing pH & temperature. In the table above, the un-ionized form is expressed as a percentage of the total ammonia. The coloration indicates the approximate toxicity of a 1 ppm ammonia level to healthy Koi. From Roark’s web site: www.click2roark.com


http://www.click2roark.com/

Table #2

Solubility of oxygen in water in milligrams per liter Atmospheric Pressure, mmHg

Temp F

Temp C 760 730 700 670 640 610 32.0 0 14.6 14.0 13.4 12.8 12.3 11.7 33.8 1 14.2 13.6 13.1 12.5 11.9 11.4 35.6 2 13.8 13.3 12.7 12.2 11.6 11.1 37.4 3 13.4 12.9 12.4 11.8 11.3 10.8 39.2 4 13.1 12.6 12.0 11.5 11.0 10.5 41.0 5 12.7 12.2 11.7 11.2 10.7 10.2 42.8 6 12.4 11.9 11.4 10.9 10.4 9.9 44.6 7 12.1 11.6 11.1 10.7 10.2 9.7 46.4 8 11.8 11.3 10.9 10.4 9.9 9.5 48.2 9 11.5 11.1 10.6 10.2 9.7 9.2 50.0 10 11.3 10.8 10.4 9.9 9.5 9.0 51.8 11 11.0 10.6 10.1 9.7 9.2 8.8 53.6 12 10.8 10.3 9.9 9.5 9.0 8.6 55.4 13 10.5 10.1 9.7 9.3 8.8 8.4 57.2 14 10.3 9.9 9.5 9.0 8.6 8.2 59.0 15 10.1 9.7 9.3 8.8 8.4 8.0 60.8 16 9.8 9.5 9.1 8.7 8.3 7.9 62.6 17 9.6 9.3 8.9 8.5 8.1 7.7 64.4 18 9.4 9.1 8.7 8.3 7.9 7.5 66.2 19 9.3 8.9 8.5 8.1 7.8 7.4 68.0 20 9.1 8.7 8.3 8.0 7.6 7.2 69.8 21 8.9 8.5 8.2 7.8 7.5 7.1 71.6 22 8.7 8.4 8.0 7.7 7.3 7.0 73.4 23 8.6 8.2 7.9 7.5 7.2 6.8 75.2 24 8.4 8.0 7.7 7.4 7.0 6.7 77.0 25 8.2 7.9 7.6 7.2 6.9 6.6 78.8 26 8.1 7.8 7.4 7.1 6.8 6.4 80.6 27 7.9 7.6 7.3 7.0 6.6 6.3 82.4 28 7.8 7.5 7.2 6.8 6.5 6.2 84.2 29 7.7 7.3 7.0 6.7 6.4 6.1 86.0 30 7.5 7.2 6.9 6.6 6.3 6.0

Source: USGS website = http://water.usgs.gov/software/DOTABLES/


http://water.usgs.gov/software/DOTABLES/

Table #3 Bagging Survey Results Each of the 4 authors of this report talked to several Koi Farms to ask them questions about the guidelines they followed when shipping Koi, the average size of shipping bags they used, and the number of each size Koi they would put in a bag. The farms were asked if they used or would recommend the use of bag additives, and if they had any advice for hobbyists when transporting Koi. Below is a compilation of what the farms said. Farm Bag Size Barstow Koi Farm 48 Brett’s Koi Farm 13x13x24 Hammock Koi Farm 20 x 44 Harrison’s Koi Farm 36 x 48 Kodama Koi Farm 20 x 40 Koda Koi Farm (Japan) 30 Koi Acres 42 Morning Light Koi Farm 24 x 48 Mystic Koi 12x12x28 Purdin Koi Farm 21 x 41 San Joaquin Koi Farm 20 x 48 All of the breeders and dealers interviewed recommend the use of ice to keep Koi cool during shipping, as well as fasting the Koi before shipment. Eight of the farms recommended the use of ULTIMATE® as a bag additive. All Koi over 18” should be triple bagged. Oxygen is much more important than water – use as much O2 as possible, and just enough water to cover the gills. While the number of Koi per bag varies depending on the time the Koi are expected to be in the bag, in general, the breeders agreed that 10-20 small Koi (under 10”), 2-3 medium Koi (10-20”) and 1 large Koi (over 20”) could be bagged together. But if shipping price was not considered (as it would not be in transporting your own Koi to a show by car), all of the breeders would agree that bagging each Koi separately will minimize stress and damage.


Table #4 Salt Amounts to get a solution of .3% table salt in a given volume of water Gallons of Water Salt Weight in lbs for .3% Useful volumes (Table Salt) 1 gal .025 1.9 teaspoons (t.) 2 gal .05 3.8 teaspoons (or 1

Tablespoon plus <1 teaspoon

3 gal .075 5.7 teaspoons, or <2 Tablespoons (T)

4 gal .1 2.5 Tablespoons 5 gal .125 3T. + 1/2 t. 10 gal .25 1/4 c + 2 T. + 1 t. 20 gal .5 3/4 c. + 2 t. 25 gal .625 ~1 cup (c.) 50 gal 1.25 ~ 2 cups 100 gal 2.5 ~ 4 cups 500 gal 12.5 ~ 20 cups 1000 gal 25 ~ 40 cups 2000 gal 50 --- 3000 gal 75 --- 4000 gal 100 --- 5000 gal 125 --- 10,000 gal 250 --- NOTES: 1) Salt should always be dissolved completely before Koi are added to the water. Direct contact between salt and Koi skin may cause irritation and even burns. 2) The term “salt” in this paper always refers to Sodium Chloride. This may take the form of common table salt, Kosher salt, Iodized salt (the iodine in salt does not harm Koi) or large bags of rock salt. 3) Adding Sodium Chloride (salt) does not release dangerous chlorine ions into the water (which would have to be neutralized with a de-chlorinator). Calcium Chloride also does not release chlorine. They release chloride, which may be beneficial to Koi.


Table #5 Recommended Tank Loading Restrictions for Koi Shows Show Tank Loading How Many Fish Can I Put into a Show Tank? By Dick Roemer Over the years MAKC has developed a method of estimating the fish load or biomass in a show tank. The method we use is called “fish points” and are a conservative but rough approximation, based on the average size of each size category. Each Koi under 8” counts as 0.3 fish points. Each Koi 8-12” counts as 1 fish point. Each Koi 12-16” counts as 2 fish points. Each Koi 16-20” counts as 5 fish points. Each Koi 20-24” counts as 8 fish points. Each Koi 24-28” counts as 12 fish points. Each Koi over 28” counts as 18 fish points A six-foot diameter tank should not contain more than 20 fish points, 25 at a maximum. An eight-foot diameter tank should not contain more than 35 fish points, 45 at a maximum.


Product Data Sheet for ULTIMATE®




Appendix ULTIMATE® ULTIMATE® has several components. The primary ones are the ingredients for ClorAm-X® (the old AmQuel®), plus the ingredients of Stress-X® (the old NovAqua®). ULTIMATE® also buffers the water and detoxifies heavy metals. It is perhaps useful to know a bit of product history. AquaScience Research Group, Inc. created the original products AmQuel® (used to detoxify ammonia, chlorine and chloramines and neutralize heavy metals) and NovAqua® (the first polymer- based, slime-coat-replacer and water-conditioner). Kordon marketed these two products for AquaScience in the early 70s and several years later decided to sever relations with AquaScience. Kordon owns the trademarks to the product names only, and AquaScience kept the patents and proprietary formulas for the products. Kordon came up with new formulations, which they still had (and have) the right to call AmQuel® and NovAqua®. The products produced by AquaScience Research Group, Inc. that contain the original molecules are now called ClorAm-X® and Stress-X®. ClorAm-X® is available in a food-fish-safe, powdered form, and does not need to be reconstituted before it is added to Koi ponds or tanks. ClorAm-X® and Stress-X® are combined with electrolytes and a chemical that boosts alkalinity to form ULTIMATE® . AquaScience Research Group, Inc. now markets their products through other companies such as Koi Care Kennel and Hikari. (information for this paragraph came from conversations and e-mail with J.F. Kuhns, the owner of AquaScience Research Group, Inc. [49]) There may be other products that are as safe and effective as ULTIMATE®. They may be substituted for ULTIMATE®, and users should follow the dosing instructions on the labels. At the time of the writing of this paper, the authors could find no scientific tests that had been performed on the other products, so their names are not included. When other research has been published, an update to this paper may include those results. Koi can live in pure ULTIMATE®. Several hobbyists as well as Norm Meck (see Biographies, Appendix) have tested this fact by placing a number of Koi in a tank of pure ULTIMATE®, and added nothing but oxygen (from an air pump and air stone) for a month. During that time, the Koi were fed regularly, and no effort was made to remove any feces from the tank. While the water became opaque from the feces and solids, water quality tests continued to indicate zero ammonia (and nitrites) at the end of the month. Obviously, this would not work indefinitely without replenishing the ULTIMATE®, as eventually, all the chemical bonds would be used up, and there would be no more bonding sites available to bind ammonia as it was being produced by the Koi. Once the ULTIMATE® is used up, ammonia levels will rise to become toxic. The research done for this paper indicates that the manufacturer’s claim that ULTIMATE® reduces stress may be valid. These finding must be verified by


rigorous scientific research in order to prove them. The authors of this paper felt that enough research had been done on ULTIMATE® to warrant passing this information along to hobbyists, with the hope that these initial results will spark other research projects that will test products that may be of use to the Koi hobby. ULTIMATE® may be used for holding tanks and quarantine tanks when a fully cycled filter is not available, but should never be a substitute for a cycled filter system. The goal of a holding or quarantine tank is perfect water, in order for Koi to be observed or to recuperate from stress or illness. A newly started filter takes weeks to cycle, and during that time, ammonia stress will add to the Koi’s problems. A filter started with chemicals and heat will take a week or so to adjust to the slight difference in the ammonia produced by Koi, and will still leave trace ammonia in the water. A fully cycled filter taken from a pond may harbor parasites or diseases from the pond. Even with a filter, it is recommended to use some polymer-based slime-coat enhancer, and ULTIMATE® may be used to treat the water whenever there is a need to hold Koi for a short duration of time. The biggest problem with creating a good quarantine tank is inadequate water quality from poorly cycled filters, and many quarantine tanks are more harmful than beneficial. Using ULTIMATE® can address the needs of the Koi without the risks of an inadequate filter. Research for this paper suggests that most Koi recover from stress more quickly when ULTIMATE® is used in the water. ULTIMATE® is now the only chemical used at many Koi shows throughout the world. All holding tanks, quarantine tanks and show tanks still require the use of aeration. ULTIMATE®, ClorAm-X® and some other ammonia reduction products will give false readings with Nessler-type ammonia test kits. Those kits are characterized by scales that read from yellow to red. When using a Nessler test kit, all of the ammonia in the water will still be read. The detoxification of ammonia works by binding the molecular ammonia, NH3, to the hydroxymethane portion of the ClorAm-X® molecule. The resulting aminomethane molecule is not toxic to Koi or other fishes. ULTIMATE® must be tested with salicylate or indophenol test kits which normally have scales that read from yellow to blue-green. Salicylate or indophenol test kits read only the residual ammonia left in the water that is harmful to Koi. All forms of ammonia (both the toxic and non-toxic ammonia molecules) are equally as useful to grow colonies of bacteria to ‘start’ a filter. ULTIMATE® and ClorAm-X® are also incompatible with Winkler dissolved oxygen (DO) tests and will give false or zero readings. An indigo/carmine dissolved oxygen test kit or a dissolved oxygen meter should be used with ULTIMATE®.


Appendix Maintaining Stable pH and Calcium In a Holding, Recovery or Quarantine Tank pH A holding, Recovery or Quarantine tank typically has a high fish density (pounds of fish per gallon of water) and may have less than perfect circulation and aeration. This can lead to constantly changing pH levels and potentially a significant drop in pH that would be fatal to the fish. Maintaining a safe alkalinity level (above 100 ppm) will result in a stable pH. The best tool for this job is sodium bicarbonate (i.e. "Arm & Hammer" baking soda). Sodium bicarbonate is quite basic and will quickly raise the pH of the tank to 8.4. One pound of sodium bicarbonate per 1000 gallons of water will raise the alkalinity (KH) by 71 ppm. A gradual increase in alkalinity (20 ppm per day) is recommended because ammonia becomes rapidly toxic at increasing pH levels, even so-called "trace" amounts can cause serious harm to fish. If a large alkalinity change is needed, do so over a period of several days. This gradual increase in alkalinity can be accomplished by adding 4-1/2 ounces (128 grams) of baking soda per 1000 gallons per day until the 100 ppm level is reached. One should test daily and re-dose as needed to maintain the 100 ppm or higher alkalinity level. Calcium To aid in stress recovery, 80 ppm calcium should be maintained, as per the discussion on page 3 in the Holding Tank section. For tanks with calcium content less than 80mg/L any of the following chemicals can be used: * Calcium Chloride (generally used to melt ice) (does not release chlorine ions) * Calcium Hydroxide (Slaked Lime) * Calcium Oxide (Lime) * Calcium Sulfate (Gypsum) The dose for Calcium Chloride is 1 teaspoon per 200 US gallons. Dissolve the chemical in a 5-gallon bucket of quarantine tank water and sprinkle it throughout the tank. If spray bars are available, run them for several hours to ensure good mixing. Measure the pH and the calcium several hours later. Apply repeat doses no sooner than 12 to 24 hours apart (keeping the pH stable) until the water has reached the 80mg/L calcium content level.


Appendix Koi Food Notes Koi food should always be fresh, as some of the necessary vitamins degrade quite quickly over time [32]. The authors of this study would recommend only buying food with a manufacturing date stamped on the bag, and replacing food that is more than 6 months old. A good strategy would be to purchase small amounts of food on a frequent basis, and only buy enough food for a few months supply. Feed should be stored air-tight in a cool, dark dry place [20, 32]. Vitamins can be adversely affected by exposure to heat, air and metals [45, 32]. Feed should not be refrigerated or frozen, as the thawing process will allow condensation to occur. Damp food will quickly become moldy, and moldy food is not only unappetizing to the Koi, it can make the Koi sick from deadly toxins [27, 32] as well. If the climate is hot, and the only cool storage option is a refrigerator, then only the amount of food required for a single feeding should be removed, while the main supply remains cold. It is the temperature change that allows harmful condensation to occur. Water Borne Antibiotics The recommended way of introducing antibiotics to Koi is either by feed or injection [46] but not usually as substances dissolved into the water. Although a common practice, water borne antibiotics are inappropriately used in the aquarium trade. Bioavailability varies from drug to drug and where absorption through the gills does occur, it can be affected by water chemistry and temperature making the delivery of the drug unpredictable [47].


Appendix Web sites with information about moving large Koi: http://www.geocities.com/koifla/transporting_koi.htmhttp://www.pond-doctor.co.uk/longbaggingkoi.htmhttp://www.petsparade.co.uk/articles/?a=16http://www.fishdoc.net/faq/faq_details.php?faqId=180http://www.koicymru.co.uk/pro11.htmhttp://www.koicymru.co.uk/stocking.htm Web sites with information about quarantine procedures: http://edis.ifas.ufl.edu/FA099http://sakoia.org/Aggressive%20Quarantine%20Protocol.pdfhttp://www.akca.org/kht/health%2020051.pdfhttp://www.koivet.com/index.php?view=article&catid=41%3Akoi-how-to-information&id=64%3Akoi-quarantine-avoiding-sick-koi&option=com_content


http://www.geocities.com/koifla/transporting_koi.htm

http://www.pond-doctor.co.uk/longbaggingkoi.htm

http://www.petsparade.co.uk/articles/?a=16

http://www.fishdoc.net/faq/faq_details.php?faqId=180

http://www.koicymru.co.uk/pro11.htm

http://www.koicymru.co.uk/stocking.htm

http://edis.ifas.ufl.edu/FA099

http://sakoia.org/Aggressive%20Quarantine%20Protocol.pdf

http://www.akca.org/kht/health%2020051.pdf

http://www.koivet.com/index.php?view=article&catid=41%3Akoi-how-to-information&id=64%3Akoi-quarantine-avoiding-sick-koi&option=com_content

http://www.koivet.com/index.php?view=article&catid=41%3Akoi-how-to-information&id=64%3Akoi-quarantine-avoiding-sick-koi&option=com_content

Biographies Information on People used as information sources for

this paper Norm Meck Koi Club of San Diego Norm teaches the Pond Water Chemistry module of the KHA course Dr. Sandra Yosha, DVM, PhD

Dr. Yosha is a nationally recognized veterinarian in Koi health medicine, and is a 1986 graduate of Tufts’ School of Veterinary Medicine. Sandra now lives in Florida and works at the Auburndale and Highland Square Vet Clinics.

Burt Ballou Burt has been a certified AKCA judge since 1997, and been in the Koi hobby for over 35 years. He was the AKCA chairman 3 times, and Koi USA Features Editor. Burt teaches the Pond Building module for the KHA course.


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