LL May 2016 - Tezasvi Publicationstezasvipublications.com/LLPDF/may2016.pdf · LIVESTOCK LINE, MAY 2016 3 VOL.10 ISSUE 1 MAY 2016 INDEX OF ADVERTISEMENTS 1. Alltech Biotechnology

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3LIVESTOCK LINE, MAY 2016

VOL.10 ISSUE 1 MAY 2016

INDEX OF ADVERTISEMENTS

1. Alltech Biotechnology Pvt. Ltd. Title Cover I I

2. B.V. Bio-Corp. Pvt. Ltd. Title Cover I

3. Jaysons Agritech Pvt. Ltd. Title Cover IV

4. Polygov 4

5. Vetoquinol Title Cover III

B. Shiv Shankar - Managing PartnerB. Kishore Kumar - Media ExecutiveB. Shailajaa - Circulation ManagerJ. Upender Rao - Marketing Manager South TelanganaSathyendranath - Marketing Manager North TelanganaK. Sudarshan - Head, Designing DepartmentP.N. Nithin - Incharge - PhotographyK. Raghuramaraju - Publication Consultant (09440231211)

1. Alltech ..... 5- Press Release

2. Biological Control Of GIN ..... 6-11- Aiman Ashraf

3. Cell Culture – An Overview ..... 12-15- Maninder Singh Sheoran

4. Diseases of Fish Caused ..... 16-17- Dr.Phaniraj.K.L

5. Effect of Mycotoxins 18-19- Raju Kushwaha

6. General Guidelines ..... 20-23- Tawheed Ahmad Shafi

7. Helmimthiasis in Fish ..... 24-28- Dr.Phaniraj.K.L

8. Homeopathic Approach ..... 29- Saraswat Sahoo

9. Internal Protozoan ..... 30-31- Dr.Phaniraj.K.L

10. Mammary Gland..... 32-36- Arvind Ku. Pandey

11. Preservation of wild ..... 37-39- S. Chaurasia

12. Role of Micronutrients ..... 40-41- Raju Kushwaha

13. Stress - Its Role ..... 42-45- Supriya, S.

14. Transport Myopathy ..... 46- Aasif Ahmad Sheikh

CONTENTS

Editor : B. KALYAN KUMARAssociate Editor : B. SHIV SHANKAR

Printed, Published and Owned by B. Shiv Shankar, Printed at Karshak Art Printers, 40, A.P.H.B. Blocks, Vidyanagar, Hyderabad - 500 044. India.Published at 2-1-444/16, 1st Floor, O.U.Road, Nallakunta,Hyd-44. Editor: B. Shiv Shankar.

TECHNICAL EDITORIAL BOARD

Dr. P.K. Shukla, Jt.Commissioner Poultry, G.O.I., New Delhi.

Dr. V. RAMA SUBBA REDDY, Retd. Professor, Agrl. Uni. Hyd.

Dr. D. NAGALAKSHMI, Asst. Professor, S.V.V.U. Hyderabad.

Dr. S.T. VIROJI RAO, Sr. Scientist, AGB, S.V.V.U. Hyderabad.

Dr. M. KISHAN KUMAR, Sr. Scientist, S.V.V.U. Hyderabad.

Dr. M. KOTESWARA RAO, Vet. Asst. Surgeon, RAHTC, KMNR.

Dr. P.K. SINGH, Asst. Prof. (A.N.), Bihar Vet. College Patna.

Dr. S. NANDI, Sr. Scientist, CADRAD, IVRI, Izatnagar, U.P.

Dr. INDRANIL SAMANTA, Lecturer (Micro), WBUAFS, Kolkata.

Dr. M. KAWATRA, Sr. Manager-Bayer Animal Health, Thane (W), Mumbai.

Dr. DEVENDRA S VERMA, Tech. Mgr, Biomin Singapore B'lore.

Dr. R.K.S. BAIS, Sr. Scientist, CARI, Izatnagar, Bareilly.

Dr. VIJAY KUMAR M, Asst. Prof., Vet. College Bidar.

Dr. MD MOIN ANSARI, Asst. Prof., SKUAST, Srinagar, J&K.

Dr. AZMAT ALAM KHAN, Asst. Prof., SKUAST, Srinagar, J&K.

Dr. S K MUKHOPADHAYAY, Asst. Prof., (Vety Pathology) WBUAFS, Kolkata.

Dr. SUBHA GANGULY, Scientist, AICRP-PHT, Kolkata Centre.

Dr. MD.MOIN ANSARI, Asst. Professor/Scientist (Sr Scale), SKUAST, Srinagar, J&K.

Dr. AIJAZ AHMED DAR, Ph.D. Scholar, IVRI, Izatnagar, Bareilly.

Dr. SARADA PRASANNA SAHOO, Ph.D. Scholar, IVRI, Izatnagar.

Livestock Line may not necessariltysubscribe to the views expressed in the Articles

published herein.

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4LIVESTOCK LINE, MARCH 2016


Alltech agrees to acquire Keenan,Ireland’s leading farming solutions manufacturer

Establishes a comprehensive Irish-based, globally-minded farming solutions and animal nutrition offering

Accelerates product innovation through combined technology, research and on-farm machineryproduction, delivering greater value to farmers

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[DUNBOYNE, Ireland] – Alltech has agreed to acquire Keenan,a leading farming solutions company in Ireland. Keenan, nowpart of the Alltech family of companies, is the 14th acquisitionfor Alltech globally since 2011.

“The Keenan group has long been a friend of Alltech. This isa story about two great Irish; globally-minded companiescoming together. As an Irishman, I am delighted to welcomeKeenan to our family, because together we can deliver greatervalue to our global farming customers with a wider variety oftechnological solutions,” said Dr. Pearse Lyons, founder andpresident of Alltech. “Between Alltech’s primacy in scienceand Keenan’s manufacturing strength and technologicalknow-how, we have a winning combination for deliveringgreater farm efficiency and profitability direct to our farmingcustomers.”

Alltech and Keenan have identified possible growthopportunities together, which may include nutritionaltechnologies and feeding programmes focused on feedefficiency and herd health as well as advanced rationformulation.

“This is an exciting time for Keenan to join us here at Alltech,”stated Alric Blake, CEO of Alltech. “Alltech is looking foravenues to better deliver the Alltech brand to farmers andprovide nutritional solutions to those who directly benefitfrom their use, whether in animal or crop production. Scienceand technology are at the forefront of everything we do. Thisnew journey with Keenan further strengthens our ability todeliver on-farm nutrition solutions.”

Keenan will continue to be headquartered in Borris, Co.Carlow, Ireland. Together, Alltech and Keenan employ nearly300 people in Ireland and close to 5,000 globally.

About Alltech:

Founded in 1980 by Irish entrepreneur and scientist Dr. PearseLyons, Alltech improves the health and performance of people,animals and plants through nutrition and scientific innovation,particularly yeast-based technology, nutrigenomics and algae.With nearly 100 manufacturing sites globally, Alltech is theleading producer and processor of yeast and organic traceminerals, and its flagship algae production facility in Kentuckyis one of only two of its kind in the world.

The company’s guiding ACE principle seeks to developsolutions that are safe for the Animal, Consumer and theEnvironment and is actively supported by close to 5,000 teammembers worldwide.

Alltech is the only privately-held company among the topfive animal health companies in the world. This is a source ofcompetitive advantage, which allows Alltech to adapt quicklyto emerging customer needs and to stay focused on advancedinnovation and long-term objectives. Headquartered justoutside of Lexington, Kentucky, USA, Alltech has a strongpresence in all regions of the world. For further information,visit www.alltech.com/news.

About Keenan:

Established in 1978, Keenan is a respected leader in ethicaland profitable farming solutions, focused on maximising feedefficiency. Over the course of nearly four decades in business,Keenan has earned a particularly strong reputation formanufacturing quality mixer wagons.

Keenan interprets data for more than 1,000,000 cows fromclose to 10,000 farms in 25 countries around the world,representing one of the world’s largest field databases ondairy feed efficiency.

Keenan prides themselves on continuous investment into newtechnology for the benefit of their customers globally. Thecompany has evolved throughout the years, combiningcutting-edge technological developments with breakthroughnutritional expertise.

A keen advocate for environmental sustainability, Keenanhas developed a range of solutions to enable farmers toovercome agricultural production challenges, improve rumenhealth and feed efficiency. Based in Borris, Co. Carlow, Ireland,Keenan employ 204 people (169 in Ireland).

For further information, visit www.keenansystem.com

The Keenan farm mixerwagon, also known as a“green machine,” hasearned a reputation for itsreliability and service.Keenan mixer wagons,together with InTouchtechnology are designed todeliver the optimal on farmfeed mix consistently.Alltech confirmed itsacquisition of Keenan;Keenan is the 14th

acquisition for Alltechglobally since 2011.

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Biological Control Of GIN Of LivestockWith Special Reference To Nematophagous Fungi

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The helminth parasites of veterinary importance belongto two phyla namely Platyhelminthes & Nemathelminthes.These parasites cause serious infections in ruminants. Theyutilize host’s nutrients and non nutrient substances and alsodamage host tissues causing mortality, morbidity andproduction losses in animals.

The advent of various anthelmintics and their relativelyeasy availability has brought about a sort of revolution incontrolling parasitic diseases, but there have been seriousconcerns about the use of these chemicals like anthelminticresistance, residual effects and also the popularity of organicfarming. India is slowly emerging as the resistance epicenterof South Asia (Sanyal, 1998). The global tempo ofdevelopment and extent of anthelmintic resistance inhelminths of small ruminants in particular, indicates that thestrategies developed and implemented over the period of last40-50 years have been incorrectly applied (Van Wyk, 2001).So, in order to become practically & ecologically sustainable,parasite control schemes need to be based on the principlesof integrated pest management (Waller, 1993). Towards thisobjective, significant advances have recently been made in:

1. Development of worm vaccines

2. Breeding of animals for parasite resistance and

3. Biological control exploiting predacious fungi

Present status of different control strategies

1. Chemical Control

Three crucial reasons for opting alternative parasitecontrol strategies including biological control are drugresistance, residues in food and environmental degradation.Frequent and haphazard use and over-reliance on chemicalsare the causes of the drug resistance. The benzimidazolesand their prodrugs are subjected to close scrutiny becausesome are known teratogens. There is greater environmentalworry regarding the avermectins than other anthelmintics.

2. Non Chemical Control

2.1. Worm vaccines

Although vaccines have been developed against someparasitic diseases e.g., Lungworm infection but for most ofthe important parasitic diseases no vaccine is available dueto the following difficulties:

� Lack of knowledge about antigens that induce protectiveimmunity

� Inability to obtain parasitic antigen in bulk (cannot becultured in lab)

� Evasion of host’s immune response

� Extreme antigenic complexity & antigenic variation ofparasites

2.2. Grazing Management

Grazing management strategies have been demonstratedto be useful to alleviate the impact of GI nematodes in livestock(Stromberg & Averbeck., 1999; Barger., 1999). The variousstrategies of grazing management are:

i. Pasture burning

ii. Pasture rotation

iii. Alternate grazing

iv. Maintaining stocking rate

Unfortunately these strategies have not been adaptedto their full extent, perhaps due to the ease for the farmers touse drugs and secondly, the increasing demand for land.

2.3. Nutritional Management

This is now a well-established fact that supplementationof the diet with additional protein does not appear to affectinitial establishment of nematode infection but the patho-physiological consequences are generally more severe onlower planes of protein nutrition (Coop & Holmes, 1996).Although many aspects of the interaction between nutritionand helminth parasites have been established but manyfeatures remain to be examined.

1Aiman Ashraf, 2I. M. Allaie, 2Z. A. Wani and 3R. A. ShahardarDivision of Veterinary Parasitology, F.V.Sc. & A.H., SKUAST-K,

Shuhama Campus, Srinagar-190006Present address:- 1M.V.Sc. Scholar,2Asstt. Professor cum Jr. Scientist, 3Prof. and Head, Division of VeterinaryParasitology, F.V.Sc. & A.H., Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir,

Shuhama Campus -190006 (J&K). Corresponding author e-mail: [email protected]


Biological control

The first well planned and successful biological controlattempt was made in California (USA) in 1888 against an insectpest, Icerya purchasi (cottony cushion scale) usingladybeetle, Rodolia cardinalis. Biological control isoperationally defined as the action of natural enemies whichmaintain a host population at levels lower than would occurin the absence of the enemies. Biological control can be dividedinto two broad categories, namely, natural and applied. Asthe name implies, natural biological control is affected bynative (or coevolved) natural enemies in the normalenvironment. Applied biological control exists due to humanintervention. It is further divided into classical (theintroduction of exotic natural enemies) and augmentative(enhancement of natural enemies already in place). Classicalprocedures have been found to reduce the population of nontarget organisms also. So proper expertise and screening ofthese organisms is required before introducing theseorganisms in a new environment. Augmentative procedureswhich generally involve manipulation of either theenvironment or the natural enemy itself are likely to be theapproaches for biological control of nematode parasites oflivestock.

Candidates for biological control of nematode parasites

All nematode parasites of livestock have a lifecyclewhich involves not only the parasitic phase within the animalhost, but also a free-living stage on pasture. All stages arepotentially vulnerable to attack by biological control agents,but that the free-living component of the parasite’s life-cycleoffers the best promise. A large range of organisms havealready been identified that are capable of egg and larvaldestruction and some of these are likely to be commerciallyexploited in the short-term. These organisms can exert theireffects indirectly by habitat (dung) destruction or directly byusing the free-living stages as a food source.

Indirect biological control candidates

1. Birds

Certain birds seek out coprophilous arthropods as a sourceof food (McCracken,1993). In doing this they can break openand scatter large deposits of cattle and horse faeces, thusallowing much quicker desiccation of the faecal material thanwould otherwise occur in the intact pats thereby helping indestroying the feeding ground of nematode larvae. But thenumber of birds involved in this activity is unlikely to besufficiently large enough to have a measurable and consistenteffect on parasite numbers on pasture.

2. Dung beetles

Dung beetle activity is directly correlated with reduction innumber of infective nematode larvae recovered from faecesand surrounding herbage, for livestock that produce largefaecal masses, i.e. cattle and horses. These dung beetles maketunnels in the dung and convert it into balls and then bury it.The effects of dung beetles is highly influenced by prevailingweather conditions so dung beetles could not be relied uponas an aid in parasite control.

3. Earthworms

Earthworms play an important and often dominating role inthe structural decomposition and disappearance of cattledung. Different species are attracted to dung pats, after initialdisruption and aeration by dung beetles and coprophilic fliesfrom which they feed. But these earthworms proliferate incool and moist conditions only which becomes a limitationfor their use as biological control agents.

Direct biological control candidates

1. Microarthropods

The effects of dung beetles can be augmented by Macrochelidmites (Order: Mesostigmata) which are carried to dung byflies and beetles on which they are phoretic. These mitesutilise dipteran eggs and larvae as their primary source ofprey, however it has been shown that they also includenematodes in their diet and that they rely on nematodes duringthose seasons when flies do not occur. These have not beenfound reliable subsequently.

2. Protozoa

The actively feeding stages of animal parasitic nematodes arethe small (most species less than 1 mm) ephemeral, first andsecond larval stages found only in the dung. The predatorysoil amoeba Theratromyxa weberi is capable of feeding onnematodes. It flows over the nematode body and assimilatesit within 24 hours. But they are slow-moving compared withnematodes and are sensitive to low soil water potentials,conditions under which nematodes may thrive, so theeffectiveness of protozoa is reduced. All these considerationsmake it highly unlikely that protozoans will offer opportunitiesfor biological control of nematode parasites of livestock.

3. Viruses

Control of several important insect pests like Hymenopteraand Oryctes is by highly species-specific forms ofBaculovirus. Consequently, although important viralpathogens against free living stages of animal parasiticnematodes are likely to exist, but formidable hurdles like


identification of such viruses and their usage method are strongdisincentives in the pursuit of this form of biological control.

4. Bacteria

Certain bacteria like Myxobacteria spp. & Pasteuria(bacillus) penetrans have been shown to be effective agentsof biological control. Myxobacteria spp. only affect rhabditidnematodes, and have potential applications in animalnematode control while as Pasteuria (bacillus) penetrans iseffective against plant nematodes only. Thus utilisation ofbacteria is still very juvenile idea in livestock farms andrequires experimental studies & trials before being applied infarms.

5. Fungus

Although all agents of biological control are of intrinsicinterest, it is within fungi that an effective and commerciallyacceptable biological control agent for nematode parasites islikely to be found. Fungi that exhibit anti-nematode propertieshave been known for a long time. They consist of a greatvariety of species, which include nematode-trapping(predacious) fungi, endoparasitic fungi and fungi that invadenematode eggs.

SOURCES OF NEMATOPHAGOUS FUNGUS

Nematophagous fungi have been found in all regions of theworld and have been reported from agricultural, garden and

forest soils, and are especially abundant in soils rich in organicmaterial. A simple method of obtaining nematophagous fungiis to use the soil sprinkling technique in which approximately1g of soil is sprinkled on the surface of a water agar platetogether with a suspension of nematodes added as a bait.The plates are observed for 5–6 weeks under a microscope atlow magnification and examined for trapped nematodes,trapping organs and conidia of nematophagous fungi.

CULTURING OF FUNGUS

Nematophagous fungi can be cultured on 2% corn mealagar (CMA) plates containing 0.02% tetracycline.

METHODS OF USING FUNGUS

1. Application to animal bedding and dung pats: In thismethod fungal chlamydospores are applied to the animalbedding and dung pats which are the grounds of larvalgrowth (Waller & Taira.,1994).

2. The best method is to allow animal to feed on fungus byincorporating fungus into animal feed. The fungus mustbe able to pass through the gut of the animal so that itcomes out viable with the faeces and is able to controllarval growth outside (Gronvold et a1.,1993).

3. Another method has been developed in which feedblocks are created which contain chlamydospores. Thismethod is also fairly effective.

MECHANISM OF ACTION

1. PREDACEOUS FUNGI: These form specialized mycelial

structures i.e. traps in the form of nets, branches, knobs or

rings (constricting) or (non-constricting) and attack

nematodes either by adhesion or mechanically.

a. Recognition and host specificity

The question of how nematophagous fungi recognize

their prey is complex. It appears that there are recognition

events in the cell–cell communication at several steps of the

interaction between fungus and nematode, which might elicit

CLASSIFICATION OF NEMATOPHAGOUS FUNGI

TYPE OF FUNGUS EXAMPLES TARGET NEMATODES

PREDACEOUS Arthobotrys oligospora, A. superba, Cooperia, Dictyocaulus,

A. conoides, A. totor, Strongyloides H.contortus, T.colubriformis,

Monacrosporium eudernatum, Oesophagostomum spp.,

Duddingtonia flagrans H.placei, O.ostertagi, Cooperia spp.,

Dictyocaulus spp., H.contortus,

T.colubriformis, Nematodirus spp.

ENDOPARASITIC Drechmeria coniospora, Ostertagia spp., H.contortus,

Harposporium anguillulae T.colubriformis

EGG PARASITIC Paecilomyces lilacinus, Ascaris spp., Trichuris spp.,

Verticilium clamydosporium Nematodirus spp.


a defined biochemical, physiological or morphological

response.

b. Attraction

Nematodes are attracted by compounds released from

the mycelium and traps of nematode-trapping fungi, and the

spores of endoparasites. Fungi that are more parasitic appear

to have a stronger attraction than the more saprophytic ones.

c. Adhesion

The three-dimensional nets are surrounded by a layer of

extracellular fibrils even before the interaction with the

nematodes. After contact, these fibrils become directed

perpendicularly to the host surface, probably to facilitate the

anchoring and further fungal invasion of the nematode.

d. Penetration

The adhesion of the traps to the nematode results in a

differentiation of the fungi. A penetration tube forms and

pierces the nematode cuticle. This step probably involves

both the activity of hydrolytic enzymes solubilizing the

macromolecules of the cuticle (Ahman et al., 1996) and the

activity of a mechanical pressure generated by the penetrating

growing fungus. The nematode cuticle is composed mainly

of proteins including collagen, and several proteases have

been isolated from nematophagous fungi that can hydrolyse

proteins of the cuticle.

e. Digestion and Storage of Nutrients

Following penetration, the nematode is digested by the

infecting fungus. Once inside the nematode, the penetration

tube swells to form a large infection bulb (Veenhuis et al.,

1985). The development of the bulb and trophic hyphae occurs

in parallel with dramatic changes in the ultrastructure and

physiology of the fungus. Lectin (Rosen et al., 1997) and

lipid droplets are involved in the assimilation and storage of

nutrients obtained from the infected nematode.

Constricting Rings

The trapping mechanism of constricting rings is

completely different. When a nematode moves into the ring,

it triggers a response such that the three cells composing the

ring rapidly swell inward and close around the nematode. The

reaction is rapid (0.1 s), irreversible, and is accompanied by a

large increase in cell volume leading to an almost complete

closure of the aperture of the trap. Following capture, the

fungus produces a penetration tube through nematode cuticle.

Inside the nematode a small infection bulb is formed from

which trophic hyphae develop.

2. ENDOPARASITES: These are the obligate parasites of

nematodes having least capacity to grow outside the body of

the host. They produce spores which attack nematodes either

by adhesion or following ingestion by the nematodes.

Endoparasites with encysting zoospores (motile spores):

They produce spores which are ingested by the nematodes.

Because of their special shapes, the spores get stuck in the

oesophagus and from there initiate infection of the nematodes

e.g Harposporium anguillulae.

Endoparasites with adhesive spores (non-motile spores): They

produce non-motile spores which adhere to, penetrate and

infect passing nematodes in the soil e.g Drechmeria

coniospora.

3. EGG PARASITES: These attack non motile stages of

parasites i.e. eggs and are thus effective against those

parasites which have a long survival time within the egg

outside the body of host in the environment. Nematodes which

lay their eggs in groups (sedentary parasites) are more

vulnerable than migratory forms. Hyphae of egg parasitic

fungi grow towards the eggs and appressoria (thickened and

flattened structure) are formed on the hyphal tips which

penetrate the egg shell. The fungi then digest the contents of

the egg, both immature and mature (containing juveniles) egg.

These egg parasites are less effective once there is a juvenile

within the egg. Fungus has chitinase enzyme, which digests

the chitin present in egg shells but not in cuticle and thus it is

able to affect eggs only and not the nematode (Lysek et al.,

1987).

CURRENT SCENARIO OF FUNGUS AS BIOLOGICAL

CONTROL CANDIDATE

Currently, the work on biological control of nematode parasites

of livestock is almost exclusively associated with the nematode

destroying microfungus, Duddingtonia flagrans. The reason

for this is that it has three very important attributes:

1. The ability to survive gut passage of livestock.

2. A propensity to grow rapidly in freshly deposited

dung.

3. Possesses a voracious nematophagous capacity.

Control is effected by the fungus capturing the infective larval

stages before they migrate from dung to pasture to complete


their life cycle following ingestion by grazing animals. Field

evaluation of this concept for a range of livestock species in

a variety of geo-climatic regions, has been underway for the

last decade and a number of potential stumbling blocks on

the path towards product registration have largely been

overcome. Firstly scaling-up of production of D. flagrans to

produce commercial quantities of spore material is possible

(Gillespie, 2002). Secondly, using D. flagrans as a nematode

control agent has no adverse effects on the environment

(Yeates et al., 1997; Yeates et al., 2002; Faedo et al., 2002;

Knox et al., 2002; Yeates et al., 2003; Waller et al., 2005).

Thirdly, it has been established that D. flagrans is ubiquitous

and that very close genetic similarity exists between isolates

from widely separated localities (Faedo, 2001; Skipp et al.,

2002), suggesting a clonal population worldwide. The

commonly used means of deployment of D. flagrans spore

material is by a feed additive. To achieve optimal results, the

fungal spores need to be continuously shed in the dung of

animals at the same time that contamination of pasture with

parasite eggs occurs. Thus daily supplementation of fungal

material is recommended during the predetermined period of

time that biological control is to be effected.

ADVANTAGES OF BIOLOGICAL CONTROL

� Biological control is an economically sustainable

method.

� All the methods of biological control have very

little or no side effects.

� Agents of biological control are self perpetuating.

� The biological control methods once established

are permanent.

� They leave no residual effect unlike chemical

agents.

� They do not interfere with the concept of organic

farming which is in much vogue nowadays.

DISADVANTAGES

� It requires subsequent use of parasiticides as it is

not efficient enough to remove the severe

infections.

� The biological control methods take time to get

established thus they act very slowly as compared

to other methods.

� The biological control methods are very

unpredictable as they have still not been

deciphered completely; their effect can be erratic

being highly effective at one place and not as

effective at another similar place.

CONCLUSION

For the vast majority of grazing livestock industries,

anthelmintics will always remain the cornerstone of effective

parasite control programmes. However, the issue of

anthelmintic resistance in nematode parasites, particularly in

small ruminants, is becoming an increasingly urgent problem.

The use of grazing management strategies, combined with

anthelmintic treatment, may well result in better parasite

control at less cost, but may not reduce significantly the

selection pressure for the development of resistance.

Development of anthelmintic resistance is testimony to the

remarkable biological plasticity of nematode parasites. Just

as strains of parasites have evolved to survive one type of

selection pressure (e.g., anthelmintics), then they are perfectly

capable of doing the same with biological control. There are

several documented cases of failure of sheep–cattle alternate

grazing strategies despite initial success of this strategy.

Biological control is not sufficiently advanced to have been

extensively used in long-term field trials for this to emerge,

but it is reasonable to assume that strains of parasites could

be selected whose free-living stages could avoid capture by

D. flagrans. Thus it is important to recognise that any specific

worm control strategy, whether it is chemotherapeutic or non-

chemotherapeutic, will be unsustainable when used in

isolation. Parasite control schemes that integrate as many

different control methods that are practically, financially and

economically feasible are the only way to ensure long-term

sustainability. Within this objective, grazing management

strategies and biological control are important components

for future parasite control schemes in grazing livestock

enterprises throughout the world.

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FAO Animal Production and Health Paper 141, FAO, Rome,

pp.54-65.

Skipp, R.A., Yeates, G.W., Chen, L.Y. and Glare, T.R., 2002.

Occurrence, morphological characteristics and ribotyping of

New Zealand isolates of Duddingtonia flagrans, a candidate

for biocontrol of animal parasitic nematodes. New Zealand

Journal of Agricultural Research., 45: 187-196.

Stromberg, B.E. and Averbeck, G.A., 1999. The role of parasite

epidemiology in the management of grazing cattle.


Van Wyk, J.A. 2001. Refugia- overlooked as perhaps the most

potent factor concerning the development of anthelmintic

resistance. Onderstepoort Journal of Veterinary Reseach.,

68: 55-67.

Veenhuis, M., Nordbring-Hertz, B. and Harder, W., 1985. An

electron-microscopical analysis of capture and initial stages

of penetration of nematodes by Arthrobotrys

oligospora. Ant v Leeuwenhoek., 51: 385-3398.

Waller, P. J., 1993. Towards sustainable nematode parasite

control of livestock Veterinary Parasitology., 48: 295-309.

Waller, P.J., Schwan, O., Ljungstrom, B.L., Rydzik, A. and

Yeates, G.W., 2005. Evaluation of biological control of sheep

parasites using Duddingtonia flagrans under commercial

farming conditions on the island of Gotland,Sweden.

Veterinary Parasitology.,126: 299-315.

Waller, P.J. and Taira, N., 1994. Proceedings Japanese Society

of Veterinary Science Annual Conference,Tokyo, p.136

Yeates, G.W., Waller, P.J. and King, K.L., 1997. Soil nematodes

as indicators of the effect of management on grasslands in

the New England Tablelands (NSW): effect of measures for

control of parasites of sheep. Pedobiologia., 41: 537-548.

Yeates, G.W., Dimander, S.O., Waller, P.J. and Höglund, J.,

2002. Environmental impact on soil nematodes following the

use of either ivermectin sustained release boluses or the

nematophagous fungus Duddingtonia flagrans to control

nematode parasites of cattle in Sweden. Acta Agriculturae

Scandinavica Section A - Animal Science., 52: 233-242.

Yeates, G.W., Dimander, S.O., Waller, P.J. and Höglund, J.,

2003. Soil nematode populations beneath faecal pats from

grazing cattle treated with the ivermectin sustained-release

bolus or fed the nematophagous fungus Duddingtonia

flagrans to control nematode parasites. Acta Agriculturae

Scandinavica Section A - Animal Science., 53: 197-206.

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CELL CULTURE – An OverviewU

What is Cell Culture?

Cell culture refers to the removal of cells from an animal orplant and their subsequent growth in a favorable artificialenvironment. The cells may be removed from the tissuedirectly and disaggregated by enzymatic or mechanical meansbefore cultivation, or they may be derived from a cell line orcell strain that has already been established.

TYPES OF CELL CULTURE

Primary Culture

Primary culture refers to the stage of the culture after the cellsare isolated from the tissue and proliferated under theappropriate conditions until they occupy all of the availablesubstrate. At this stage, the cells have to be sub cultured(i.e., passaged) by transferring them to a new vessel withfresh growth medium to provide more room for continuedgrowth..Primary cell culture could be of two types dependingupon kind of cells in culture-Adherent cell and Suspensioncells. Adherent cell - cells shown to require attachment forgrowth are said to be anchorage dependent cells. Suspensioncells-cells which do not require attachment for growth or don’tattach to the surface of the culture vessels.

Secondary culture

When a primary culture is subcultured it becomes secondaryculture or cell line.subculture (passage) refers to the transfer

of cells from one culture vessel to another culture vessels.

Cell Line :After the first subculture, the primary culturebecomes known as a cell line or sub-clone. Cell lines derived

from primary cultures have a limited life span (i.e., they arefinite)

Finit vs Continuous Cell Line

Normal cells usually divide only a limited number of timesbefore losing their ability to proliferate, which is a geneticallydetermined event known as senescence; these cell lines areknown as finite. However, some cell lines become immortalthrough a process called transformation, which can occurspontaneously or can be chemically or virally induced. Whena finite cell line undergoes transformation and acquires theability to divide indefinitely, it becomes a continuous cellline.

Culture Conditions

Culture conditions vary widely for each cell type, but theartificial environment in which the cells are cultured invariablyconsists of a suitable vessel containing the following:

• a substrate or medium that supplies the essentialnutrients (amino acids, carbohydrates, vitamins,minerals)

*Maninder Singh Sheoran1, Sandeep Kumar2, Sandeep Gera3 ,C.S Patil2, Kapil Dev1

1Mvsc Scholar, 2Assistant Professor, 3Professor ,Department of Veterinary Physiology And Biochemistry, LUVAS Hisar

*Corresponding Author: [email protected]


• growth factors

• hormones

• gases (O2, CO

2)

• a regulated physico-chemical environment (pH,osmotic pressure, temperature)

Types of Cell Culture Media

Animal cells can be cultured eitherusing a completely natural mediumor an artificial/synthetic mediumalong with some natural products.

Media Type Examples Uses

Biological Fluids plasma, serum, lymph, human placental cord serum,

amniotic fluid

Natural media Tissue Extracts Extract of liver, spleen, tumors,

leucocytes and bone marrow,

extract of bovine embryo and chick embryo

Clots coagulants or plasma clots

Balanced salt PBS, DPBS, HBSS, EBSS Form the basis of

solutions complex media

Artificial media Basal media MEM DMEM Primary and

diploid culture

Complex media RPMI-1640, IMDM Supports wide range

of mammalian cells

Table 1. Types of natural and artificial media.

Natural Media

Natural media consist solely of naturally occurring biological

fluids. Natural media are very useful and convenient for a

wide range of animal cell culture. The major disadvantage of

natural media is its poor reproducibility due to lack of

knowledge of the exact composition of these natural media.

Artificial Media

Artificial or synthetic media are prepared by adding nutrients

(both organic and inorganic), vitamins, salts, O2 and CO

2 gas

phases, serum proteins, carbohydrates, cofactors .

Artificial media are grouped into four categories:

Serum containing media

Fetal bovine serum is the most common supplement in animal

cell culture media. It is used as a low-cost supplement to

provide an optimal culture medium. Serum provides carriers

or chelators for labile or water-insoluble nutrients, hormones

and growth factors, protease inhibitors, and binds and

neutralizes toxic moieties.

Serum-free media

Presence of serum in the media has many drawbacks and can

lead to serious misinterpretations in immunological studies.

A number of serum-free media have been developed . These

media are generally specifically formulated to support the

culture of a single cell type and incorporate defined quantities

of purified growth factors, lipoproteins, and other proteins,

which are otherwise usually provided by the serum. These

media are also referred to as ‘defined culture media’ since the

components in these media are known .

Chemically defined media

These media contain contamination-free ultra pure inorganic

and organic ingredients, and may also contain pure protein

additives, like growth factors . Their constituents are

produced in bacteria or yeast by genetic engineering with the

addition of vitamins, cholesterol, specific amino acids, and

fatty acids.

Protein-free media

Protein-free media do not contain any protein and only contain

non-protein constituents. Compared to serum-supplemented


media, use of protein-free media promotes superior cell growth

and protein expression and facilitates downstream purification

of any expressed product. Formulations like MEM, RPMI-

1640 are protein-free and protein supplement is provided when

required.

Morphology of Cells in Culture

Cells in culture can be divided in to three basic categories based on their shape and appearance .

Fibroblastic (or fibroblast-like) cells are bipolar or multipolar, have elongated shapes, and grow attached to a substrate.

Epithelial-like cells are polygonal in shape with more regular dimensions, and grow attached to a substrate in discrete

patches.


U

Lymphoblast-like cells are spherical in shape and usually

grown in suspension without attaching to a surface.

Applications of cell culture

Cell culture is one of the major tools used in cellular andmolecular biology, providing excellent model systems forstudying the normal physiology and biochemistry of cells(e.g., metabolic studies, aging), the effects of drugs and toxiccompounds on the cells and mutagenesis and carcinogenesis.The major advantage of using cell culture for any of theseapplications is the consistency and reproducibility of resultsthat can be obtained from using a batch of clonal cells.

I. Model System :Cell culture are used as model system tostudy basic cell biology and biochemistry, to study theinteraction between cell and disease causing agents likebacteria, virus, to study the effect of drugs, to study theprocess of aging and also it is used to study triggers forageing.

II. Cancer Research : The basic difference between normalcell and cancer cell can be studied using animal cellculture technique, as both cells can be cultured inlaboratory. Normal cells can be converted into cancercells by using radiation, chemicals and viruses. Cellculture can be used to determine the effective drugs forselectively destroy only cancer cells.

III. Virology : Animal cell cultures are used to replicate theviruses instead of animals for the production of vaccine.Cell culture can also be used to detect and isolateviruses, and also to study growth and development cycleof viruses. It is also used to study the mode of infection.

IV. Toxicity Testing : Animal cell culture is used to studythe effects of new drugs, cosmetics and chemicals onsurvival and growth of a number of types of cells.Especially liver and kidney cells. Cultured animal cellsare also used to determine the maximum permissibledosage of new drugs.

V. Vaccine Production : Cultured animal cells are used inthe production of viruses and these viruses are used toproduce vaccines. For example vaccines for deadlydiseases like polio, rabies, chicken pox, measles andhepatitis B are produced using animal cell culture.

VI. Genetically Engineered Protein: Animal cell culturesare used to produce commercially important geneticallyengineered proteins such as monoclonal antibodies,insulin, hormones, and much more.

VII. Replacement Tissue or Organ: Animal cell culture canbe used as replacement tissue or organs. For exampleartificial skin can be produced using this technique totreat patients with burns and ulcers. However researchis going on artificial organ culture such as liver, kidneyand pancreas. Organ culture techniques and researchare being conducted on both embryonic and adult stemcell culture. These cells have the capacity to differentiateinto many different types of cells and organs.

VIII. Genetic Counseling: Fetal cell culture extracted frompregnant women can be used to study or examine theabnormalities of chromosomes, genes usingkaryotyping, and these findings can be used in earlydetection of fetal disorders.

IX. Genetic Engineering: Cultured animal cells can be usedto introduce new genetic material like DNA or RNA intothe cell. These can be used to study the expression ofnew genes and its effect on the health of the cell. Insectcells are used to produce commercially importantproteins by infecting them with genetically alteredbaculoviruses.

X. Gene Therapy: Cultured animal cells can be geneticallyaltered and canbe used in gene therapy technique. First cells areremoved from the patient lacking a functional gene ormissing a functional gene. These genes are replaced byfunctional genes and altered cells are culture and grownin laboratory condition. Then these altered cells areintroduced into the patient.

XI. Drug Screening and Development: Animal cell culturesare used to study the cytotoxicity of new drug. This isalso used to find out the effective and safe dosage ofnew drugs. Now these tests are being conducted in 384and 1536 well plates. Cell-based assay plays an importantrole in pharmaceutical industry.

Conclusion : A single cell is the building block for life. Thegenetic material of each cell in the body - itself composed of100 trillion cells - holds the secret to inherited diseases, suchas Tay Sachs, cystic fibrosis and other complex diseases likeheart disease. Tissue culture is free of the variations thatmight arise in the whole organism - in response to normal andinduced experimental stress. But now cell culture techniqueplays an important role in research and development of drugdiscovery and also helps in improving the health and qualityof life of patients suffering from dangerous diseases like cancer,genetic disorders.


DISEASES OF FISH CAUSED BY FUNGUSU

Fungal infections (fungal infections are called mycoses)

are among the most common diseases seen in temperate fish.

Because fungal spores are found in all fish ponds and create

problems in stressed fish. Possibility of fungal infections are

Poor quality of water, Poor hygiene, Fish that are injured have

other diseases, Dead fish/large amounts of decomposing

organic material in the pond. Poor water quality can also lead

to an increase in fungal infections in an otherwise healthy

fish population. Most fungal infections only attack the external

tissues and only few fungal infections that will infect the

internal organs of fish.

Saprolegniasis

Saprolegnia can act as a primary pathogen infecting fish

that have not shown signs of previous damage. This diseases

is also called as Cotton wool fungus disease, The most common

presentation of water mold infection as relatively superficial,

cotton like growth on the skin or gills. Such lesions usually

begin as small, focal infections that can rapidly spread over

the surface of the body. New lesions are white and over time

will become red, brown, or green.

This disease attacks are temperature-dependant (temperature

ranging from 32° to 95°F but seem to prefer 59° to 86°F) usually

occurring at low temperatures.

A. Caused by various groups of aquatic fungi; primarily

Saprolegnia, Achlya, and Aphanomyces.

B. Saprolegniasis affects all species and ages of freshwater

and estuarine fish.

C. Clinically, affected fish develop white to brown cotton

like growths on skin, fins, gills and dead eggs. This

organism is an opportunist that will usually grow over

previous ulcers or lesions. Diagnosis is by finding broad

nonseptate branching hyphae that produce motile

flagellated zoospores in the terminal sporangia.

D. In the Atlantic menhaden, gizzard shad, and some other

marine fishes, this fungus may present as an ulcerative

mycosis that may progress to a deep necrotic lesion

involving the muscle. Histologically there is an intense

granulomatous inflammation with broad (7 to 14 micron),

nonseptate hyphae.

E. Most fish die due to osmotic or respiratory problems if

the affected area of skin or gills is large.

F. The fungi are normal water inhabitants that invade the

traumatized epidermis. Improper handling, bacterial or

viral skin diseases, and trauma are the major causes of

the disease. It is interesting to note that temperature

has a significant effect on the development of infections.

Most epizootics occur when temperatures are below the

optimal temperature range for that species of fish.

Saprolegniasis is mainly a secondary infection seen after

damage to the fish integument. Water pollution and

overcrowding like other predisposing factors were also

include. George, et al. (1998) reported that typical

saprolegniosis lesions grow surface of the skin, they usually

do not penetrate deeply into muscle. The area of skin and gill

damage determines the severity of the disease.

The disease show symptoms like -Fish fungus appears

as gray or white patches on the skin/gills, they may become

brown/green (later stage) as they trap sediment and

Saprolegnia normally establishes as small, focal infections

that then spread rapidly over the body or gills.

Treatment: Fish are removed from the water they appear

to have a “slimy” matted mass growing out of the skin and

scales. Use the 100mg/ litter strong malachite green solution

to clean the lesion and apply a waterproof cream.

Branchiomycosis (Gill rot)

A. Caused by two species Branchiomyces sanguinis and

B. demigrans.

B. Primarily a problem in carp, rainbow and brown trout,

and eels.

C. Affected fish usually show respiratory distress. There

is prominent gill necrosis caused by thrombosis of blood

1Dr.Phaniraj.K.L M.V.Sc., Ph.D., and 2KISHOREKUMAR

Complete postal address: 1Assistant Professor, Department of Veterinary Microbiology, Veterinary College,Karnataka Veterinary Animal and Fisheries Sciences University. SHIVAMOGGA.

E-mail address of the corresponding author: [email protected]


vessels in the gills. Histologically, the identification of

nonseptate branching hyphae with an intrahyphal

eosinophilic round body (apleospores) in and around

blood vessels of the gill is diagnostic.

D. The disease occurs most commonly in overcrowded

ponds with abundant organic matter and high ammonia

levels. Usually warm water temperatures (20-25°C) bring

about the disease.

Ichthyosporidiosis

Gustafson and Rucker (1956) reported that Ichthyosporidium

is a fungus, but it manifests itself internally. It primarily attacks

the kidneys and liver, but it spreads everywhere else. The

disease symptoms are, The fish may become sluggish, lose

balance and eventually show external cysts or sores. For the

Treatment, can use 1% Phenoxethol solution added to food

or Chloromycetin added to the food has also been effective.

A. Ichthyophonus hoferi; large 10 250 micron spores which

may germinate to form large hyphae (similar to the

hyphae of Saprolegnia).

B. This fungus infects all species of fish.

C. Clinically the fish are emaciated with small round

occasionally ulcerated black granulomas in the skin.

Scoliosis is occasionally observed. Internally, numerous

granulomas are observed in many visceral organs.

Microscopically, the lesion consists of granulomas with

encysted large PAS positive spores. Occasionally large

irregular shaped hyphae are observed.

D. Transmission is unknown, but believed to be due to

ingestion of contaminated feed.

Exophiala sp.

A. Exophiala salmonis and E. psychrophila; these fungal

organisms have hyphae that are septated, irregular in

width and branched.

B. This disease is observed in many species of fresh and

saltwater fish. E. salmonis has become an organism of

increased importance in caged cultured salmonids.

C. Clinically the fish become darker and lethargic, with

erratic and whirling swimming behavior. Occasionally

dermal nodules are present. Numerous round yellow to

white granulomas are present in visceral organs (liver,

kidney, spleen) with prominent enlargement of the

posterior kidney common. Histologically, branched,

irregular width, septated hyphae are present in the

lesions.

D. Transmission is unknown.

Exophiala sp: Exophiala salmonis and E.psychrophila ; these

fungal organisms have hyphae

that are septated, irregular in width and branched (Robert,

1989). Both fungal diseases infected the many species of fish.

Symptoms:

1. Fish become darker and lethargic, with erratic and

abnormal swimming behavior.

2. Round yellow to white granulomas are present in visceral

organs like liver, kidney and spleen with prominent

enlargement of the posterior kidney

Reference:

1. Roberts R.J: Fish Pathology, Bailliere Tindall, London,

Second edition, 1989.

2. Ferguson H.W.: Systemic Pathology of Fish, Iowa State

Press, Ames, Iowa, 1989.

3. Anderson B.G.: Atlas of Trout Histology, Wyoming

Department of Fish and Game, 1974.

4. Fox J.C.: Laboratory Animal Medicine, Academic Press,

1984.

5. Magaki G., Rebelin W.E.: The Pathology of Fishes, The

University of Wisconsin Press, 1975.

6. Wolf K.: Fish Viruses and Fish Viral Diseases, Cornell

University Press, London 1988.

7. Tucker C.S.: Channel Catfish Culture, Elsevier Science

Publishers, Amsterdam, 1985.

8. Principal Diseases of Farm Raised Catfish, Southern

Cooperative Series Bulletin No 225, 1985.

9. Wales J.H.: Microscopic Anatomy of Salmonids. An

Atlas, United States Department of the Interior, Resource

Publication 150, 1983.

10. Grizzle J.M.: Anatomy and Histology of the Channel

Catfish, Auburn Printing Co, 1976.

U


Effect of Mycotoxins on the production performance of Dairy CowsU

Dairymen work to keep their cows healthy and productive,which again, isn’t always easy. Naturally occurring toxiccontaminants in feedstuffs, which can adversely affect animal

performance and health, are an ever present threat. Thatmycotoxins suppress the immune system and affect the normalfunctioning of major organs including the rumen, intestinal

tract, liver, kidneys, reproductive system, nervous system,etc. is well documented. Down on the dairy farm, the incidenceof diseases such as displaced abomasum, ketosis, retained

placenta, metrites, mastitis and fatty livers increases withmycotoxin exposure. Mycotoxin induced diseases seldomrespond if at all to veterinary therapy and result in increasing

losses if only veterinary solutions are pursued. Furthermore,ration adjustments and management changes (grouping, cowmovement, stall allotment, etc.) are of little value although

they may be a factor in predisposition to mycotoxicoses.Initially, mycotoxins, such as aflatoxins and trichothecenes,act on the immune system (number of macrophages,

lymphocytes and erythrocytes) reducing the animal’sresponse to challenges. At higher levels they affect rumen(reduced concentration of microorganisms, decreased rumen

motility) and other organ functions.

Another aspect that should be taken into account is thehigher incidence of lameness on dairy farms contaminatedwith mycotoxins. Lameness alone in dairy farms already causes

large financial losses due to a decreased milk production,impaired reproductive performance and higher culling andveterinary costs. In a study a positive relationship was

established between aflatoxin contamination of feed, lameness(subclinical laminitis) and impaired fertility (cystic ovaries).For these animals with a completely developed fore stomach

system, the rumen fluid content is, for certain mycotoxinssuch as ochratoxin A, zearalenone, T-2 toxin,diacetoxyscirpenol and deoxynivalenol, a detoxifying barrier

with protozoa being significantly more active than bacteria.For this reason, it is often thought that ruminants are protectedagainst the harmful effects of mycotoxins due to the action of

ruminal microorganisms. However, other aspects should betaken into account before disregarding mycotoxins’ hazardouseffects in ruminants. First of all, for some of these toxic

Raju Kushwaha*, Muneendra Kumar, Vinod Kumar, Debashis Roy, Shalini VaswaniDepatment of Animal Nutrition,

College of Veterinary Science and Animal Husbandry, DUVASU, Mathura-281001*e-mail: [email protected]

compounds namely aflatoxin and zearalenone, metabolic by-products are as toxic as or more toxic than the originalmolecules. Secondly, it should always be considered that

mycotoxins will adversely impact rumen environment andactivity even before having an effect on the animalsthemselves.

Aflatoxins

Early indications of aflatoxin toxicity include reduction

in feed intake followed by weight loss or a slower rate of gain.Also, there is usually a decline in feed efficiency, increasedsusceptibility to stress, and poor reproductive performance.

Calves are more susceptible than older animals. Chronicaflatoxicosis is characterized by unthriftines, anorexia, a dryingand peeling of skin on the muzzle, prolapse of the rectum,

liver damage, elevated levels of blood constituents and edemain the abdominal cavity. Milk production may dropdramatically in dairy cows fed aflatoxin contaminated feed.

Almost any level of aflatoxin contaminated feedstuff in theration may lead to some liver damage, especially in younganimals. Histopathological findings include cholangiectasis,loss of liver cell glycogen, fatty degeneration, fibroblastic

proliferation and perivascular edema – all of which are seen inthe liver. Metabolisation into aflatoxicol, a highly toxic aflatoxinB1 derivative, has also been detected. Low conception rate,

cystic ovaries and uterine infection were observed in dairyanimals consuming a naturally aflatoxin-contaminated diet.In the case of aflatoxins, not only the decrease in the

productivity and the impact on animal health should beconsidered. The carry-over of aflatoxin residues into the milkshould not be ignored as legislation exists worldwide limiting

the concentration of AfM1– the milk metabolite of AfB1 in milk(namely: 0.5 ppb in the USA and 0.05 ppb in the EU). AflatoxinM1 appears in the milk within hours of consumption and

returns to baseline levels within two or three days after removalof contaminated feed from the diet.

Ochratoxins

Ochratoxin A is a nephrotoxic mycotoxin formed byAspergillus and Penicillium spp.. Experimental examinations

of 30 day old calves which received 0.1-0.5 mg ochratoxin A/


kg LM (live mass) daily over a period of four weeks, showed

polyuria, depression, decreased weight gain, low specificgravity of urine and dehydration. At necropsy, grayish coloredkidneys and a mild enteritis were seen. Histopathological

findings comprised of slight tubular degeneration withabundant eosinophilic, hyalinic material as a sign of depositionof protein into the tubules and Bowmann’s capsules.

Furthermore, necrosis of the epithelium of proximal tubulesand intersticial fibrosis occurred. Ochratoxin A was also foundcombined with citrinin, a metabolic product produced by the

same fungi.

Zearalenone

Zearalenone (ZEA) is an estrogenic metabolite of several

species of Fusarium which has been reported to occur insilage, corn and other grains such as soybean, wheat, barley,oats, sorghum, sesame seed, and hay in many areas of the

world. Chemically, zearalenone shows a similar configurationto estradiol enabling it to connect to cytoreceptors, thuscausing estrogenic effects as well as abnormal estrus. More

than 90% of ingested zearalenone is known to be convertedinto á-zearalenol (about 10 times more estrogenic) in the rumenand to a lesser extent to ß-zearalenol (lower toxicity). Vulvar

mucous discharge, repeated AI, increased culling due toinfertility and difficult heat detection were observed whenanimals were fed hay and silage which tested positive for

ZEA contamination.

Fumonisins

Fumonisins are mainly produced by Fusarium

verticillioides (syn. moniliforme) as well as by Fusariumproliferatum and they occur predominately in maize and maizebased feeds. Dairy cattle fed diets containing 100 ppm

fumonisins for approximately seven days prior to fresheningand 70 days thereafter demonstrated lower milk production,explained primarily by reduced feed consumption. Higher

levels of serum enzyme concentrations found suggested liverdisease.

T-2 toxin

T-2 toxin is a very potent Type-A trichothecene,produced by Fusarium fungi. In cattle it has been associatedwith gastroenteritis, intestinal hemorrhages and death. T-2

toxin has also been related to feed refusal and gastrointestinallesions, bloody diarrhea, low feed consumption, decreasedmilk production, and absence of estrus cycles. Observations

in dairy herds affected with T-2 toxin at dietary levels of 300 to

500 ppb suggest that T-2 toxin reduces milk production,

hinders adjustment of fresh cows to the lactation diet, causesdiarrhea and intestinal irritation, and increases culling anddeath rates.

Deoxynivalenol

Deoxynivalenol (vomitoxin, DON) is a mold toxinproduced by Fusarium species. It has been associated with

reduced feed intake, unthriftiness, reduced weight gain, anddecreased performance. Other symptoms include diarrhea,abortion, hemorrhage, hematological changes, and nervous

disturbances. The impact of DON in dairy cattle is not wellunderstood, but clinical data shows an association betweenDON intake and poor performance. Deoxynivalenol may,

therefore, be a marker for low-quality mycotoxin-contaminatedfeed in these herds. Other field reports help substantiate alink between DON and poor performing dairy herds and it has

also been associated with reduced feed intake in non-lactatingdairy cattle.

Economic Impact

Contradictory data may be available in literatureconcerning mycotoxin’s toxicity. However, one should alwaysinterpret information taking into account that animal

production is a very dynamic process where many interactantfactors are present. Mycotoxins are a serious problem per se.The existence of other toxins, unbalanced nutrition, poor

hygiene, hard weather conditions and/or pathologicalproblems in the herd at the same time as mycotoxin exposure,are likely to amplify their negative effects.

Conclusion

The presence of mycotoxins in feed can hit all animalproducers hard. Loss of productivity, and sometimes loss of

the finished product can result from feeding grains with highlevels of mycotoxins. Among the most affected species arehigh producing dairy cattle. The importance of quality

feedstuffs to producers can mean the difference between profitand loss. Effectively reducing the amount of mycotoxins infeed is oftentimes critical to achieving the best production.

As it is stated in this newsletter and in other publications,many mycotoxins can impair the health and productivity ofdairy animals. Based on this information, we should wonder

why dairymen and their veterinarians do not employ amycotoxin deactivator more often than they do. Unfortunately,when they do, it is usually a last resort since nothing else has

solved their problem.

U


GENERAL GUIDELINES FOR FEEDING CANINESU

Introduction

A dog’s general physical condition can tell you a lot

about the diet it is being fed. If a dog does not receive a

nutritionally balanced diet, his general wellbeing will suffer.

We say a dog is on balanced diet if the dog’s energy level is

right for his breed and age, if his skin and coat are healthy, if

his stools are firm and brown, and if he seems to be in overall

good health. The amount of food a dog requires depends on

the animal’s age, breed, gender, activity, temperament,

environment and metabolism. Dogs exhibit omnivorous

feeding behaviour and therefore their diet should be comprised

of proteins, carbohydrates, fats, vitamins, minerals and water

in the correct proportions.

Dog food nutrients

1. Proteins

Protein often called the “building blocks” of the tissues

is essential for healthy growth and repair. Skin and muscle

tissue both contain large amounts of protein and it is also the

main component of hair and nails. In dogs, protein is also an

important energy source. Proteins are comprised of 23 different

amino acids andthe dog’s body can manufacture 13 of these

amino acids. The other 10 amino acids, however, must come

from dietary meat and plant sources and are called the

“essential amino acids”. The biological value of a protein is a

measure of that protein’s ability to supply amino acids,

particularly the 10 essential amino acids, and to supply these

amino acids in the proper proportions. In general, animal

proteins (meat, by-product meal) have higher biological value

than vegetable proteins (soybean meal, corn gluten

meal).Dietary protein can come in many forms from many

sources. The most natural and digestible form for dogs comes

in meat and fish. As with most things, when it comes to protein,

quality is much more important than quantity.

2. Fats

Fats (or oils as they are often called) serve a number of

essential functions in dogs. Healthy skin and hair are

maintained by fat and per gram, fat provides more than twice

1Tawheed Ahmad Shafi and 2Abdul Qayoom Mir1, 2PhD Scholar, Veterinary Medicine

Guru Angad Dev Veterinary and Animal Sciences University

the energy of protein or carbohydrates. Certain fats, called

essential fatty acids (commonly known as omega 3 and 6 oils)

cannot be made by the dog and therefore must be obtained

from food. These essential oils are important in controlling

inflammation, blood clotting, and brain development and too

little can lead to health problems.Fats are used to supply

energy, essential fatty acids, and transport the fat-soluble

Vitamins A, D, E and K. In addition, fats make a diet more

palatable to a dog. Fats help to maintain a healthy skin and

hair-coat. However, if a dog’s diet is very high in fat it may

result in the dog eating an excessive amount of energy that

may predispose to weight gain and obesity. The majority of

dry dog foods contain 9-14% fat (about 2-4% in wet foods). If

your dog is prone to weight gain, you should look for foods

with no more than 10% fat (2.5% wet).Common nutritious oil

supplements include fish oils, evening primrose oil, borage

oil and rosemary oil.

3. Carbohydrates

Carbohydrates are supplied in the diet from plant

sources such as grains and vegetables. Carbohydrates are a

direct source of energy and are also protein-sparing nutrients.

Without carbohydrates and fats, the dog’s body must convert

protein to glucose to obtain energy; consequently, these

proteins are no longer available for the building and

maintenance of lean body tissues.

4. Vitamins

Vitamins are a group of compounds that are essential

for keeping your dog fit and healthy. They cannot be produced

in sufficient quantities by the body and so have to be taken in

through diet and because they aren’t stored very efficiently,

daily intake is important. Fat-soluble Vitamins A, D, E and K

need fat in the diet to be absorbed by the body. The B-complex

vitamins dissolve in water and are readily absorbed by the

body. Vitamin C also dissolves in water, but it is not needed in

the canine diet because dogs can make it themselves. There

are currently 13 known vitamins, each of which serves a crucial

role in your dog’s health:


• Vitamin A: Necessary for vision, growth, immune

function, foetal development, healthy skin and coat.

• B Vitamins: Are primarily involved in metabolising, or

deriving energy, from the foods you eat.

• Vitamin C: Vital for a robust immune system.

• Vitamin D: Important during skeletal development,

phosphorus balance, necessary to absorb calcium in

the intestine.

• Vitamin E: Defense against oxidative damage.

• Vitamin K: Is involved in bone development and blood

clotting.

5. Minerals

Dogs need a wide variety of minerals to stay fit and

healthy, all of which have to be in sufficient quantities in any

complete food.Minerals are needed by the body for structural

building and chemical reactions. Minerals are involved in every

process in the dog’s body. Here’s a list of some of the most

important minerals and the roles they perform:

• Calcium: Necessary for the formation of bone and teeth,

nerve transmission, muscle contractions.

• Phosphorus: Required for skeletal structure, DNA, RNA

structure; energy metabolism.

• Magnesium: Needed to allow enzymes to function;

hormone secretions; nerve cell membrane interface.

• Potassium: Required for healthy nerve function; enzyme

reactions; energy metabolism.

• Iron: Integral part of haemoglobin and myoglobin;

energy metabolism; enzymes in respiration.

• Copper: Connective tissue; iron metabolism; blood cell

formation and defense against oxidation.

• Zinc: Enzyme function; protein and carbohydrate

metabolism; skin function and wound healing.

• Manganese: Enzyme reactions; bone development;

cartilage formation; neurological function and

metabolism.

• Selenium: Important in the immune system and

protection against oxidisation.

6. Water

Water is the most important nutrient for all animals.

Healthy dogs regulate their water intake so long as clean and

fresh water is always available. A dog can lose all its body fat

and half of its protein and survive; but if it loses only one-

tenth of its water, the dog may not survive.

7. Fibre

Although there is some discussion over whether dogs

need fibre in their diet or not, there is now a growing consensus

that dietary fibre can be very beneficial for dogs.Fibre absorbs

water like a sponge. This means that if there is excess water in

the colon, for example during diarrhoea, any dietary fibre will

soak it up and help to produce a firm stool. If, on the other

hand, there is too little water in the colon, which often leads

to constipation, the fibre will draw water in from surrounding

tissues and help to resolve the problem. As you can see, fibre

is important in maintaining intestinal health and can effectively

treat both constipation and diarrhoea.

Another important function of fibre is as a pre-biotic. This

means that is provides a medium and a food source for ‘friendly’

intestinal bacteria. These bacteria aid in the digestion of food

and help to prevent harmful bugs from getting established.

Dietary fibre also slows down the digestion of the other foods

it is consumed with. This can be particularly useful in diabetic

dogs because the fibre helps to provide a slow, steady release

of dietary sugar into the bloodstream. It can also help with

weight loss programs as foods that are high in fibre are

digested more slowly, allowing the dog to feel fuller for longer

while providing less calories.

Nutrition of dogs based on life stages

Feeding dog a nutritionally balanced diet is essential to

maintain health and vitality. The ingredients in a natural diet

vary only slightly from puppy to adulthood. They comprise

the four basic food groups: Proteins, fats, carbohydrates and

vegetables. The ratios of these in the diet will vary with the

different nutritional requirements of age (stage of growth),

metabolism, energy expenditure or exercise levels and

reproductive status. A diet based on raw meats (both muscle

meats and some organ/offal), bones, mixed cereal grains,

vegetables and fruits, and a few basic natural supplements to

ensure vitamin/ mineral balance, can be adjusted to suit all

stages of a dog’s nutritional needs.

Canine nutrition can be divided into various “life stage” viz.,

puppy, adult dog and senior dog ranges, targeting the specific


requirements of each age group. The general feeding

guidelines are given as below:

Feeding Puppies

Puppies need a clean, warm, draft-free nesting area. Air

temperature in the immediate vicinity of the puppies should

be 85° to 90° F for the first week of life, 80° F the next 3-4

weeks and 70° to 75° F at 6 weeks. Puppies need more calories

and essential nutrients than do adult dogs. Puppies under six

months should get three or four meals a day. They are growing

rapidly, but their stomachs have limited capacity. After six

months they can handle two to three meals a day.Puppies

may be fed by bottle or stomach tube. The stomach tube is

much faster and especially handy with large litters. Newborn

puppies should be fed 3-4 times daily by tube feeding or 5-6

times daily by bottle feeding. At 2 weeks of age, 3 tube

feedings or 4 bottle feedings are usually sufficient. Solid foods

should be introduced at 3 weeks of age.

The total daily caloric requirements for puppies <4 weeks of age are:

Age in weeks Caloric requirements(calories/ounce of body weight daily)

Ist 3.75

2nd 4.5

3rd 5

4th 5.5

Feeding Weanling Puppies

Weaning is the process of gradually changing a puppy’s diet

from mother’s milk to solid foods. Usually this period is from

3-4 weeks of age until 6-8 weeks of age. Early weanlings

should be weighed frequently and their weight recorded.

Progressive weight gain and content puppies are good

indicators of adequate nutrition. Begin weaning the puppies

around 3-4 weeks of age by pan feeding bitch’s milk substitute.

The first pan feedings usually consist of the puppies wading

through the food and lapping very little. Most puppies lap

from a pan readily after 3-4 feedings. When puppies are lapping

the milk substitute readily (by 3 ½ - 5 weeks of age), blend the

milk substitute and a good quality puppy food to form a thin

gruel. This should be offered to the puppies 3-4 times daily.

When the puppies are eating the thin gruel readily, the amount

of milk substitute added should be gradually reduced and the

gruel slowly thickened. The goal is to eliminate the milk

substitute by 6-7 weeks of age. At this age, the pups should

be eating good quality puppy food softened with water 3-4

times daily. Water can be eliminated when the teeth have

erupted and the pups are vigorously chewing.

Feeding Adult Dogs

The most important thing to keep in mind when feeding an

adult dog is to make sure that it is provided a complete and

balanced diet. Homemade diets can provide complete

nutrition, but making sure that pet gets the right mix of protein,

fats, minerals, and vitamins can be difficult so pet foods can

be used. The adult stage of life is usually considered as that

from around 12 months of age through to 7 years. According

to the Merck Veterinary Manual a dog is considered an adult

for feeding purposes when it reaches 90% of its expected

adult weight.

Total daily caloric requirements for dogs of different weight ranges

Dog’s Weight (lb) Caloric requirement (calories/pound of body weight)

1-2 60

3-5 52

6-10 45

11-14 40

15-29 35

30-45 30

46-74 27

75 23


An adult dog diet, or maintenance diet, contains nutrients

suited for animals that have passed their growth stage. As

an adult a dog will have his own specific nutritional needs

to keep him in peak condition and help him live a long and

active life. Factors to consider when choosing a diet include

dog’s age, activity, breed, and temperament. Also, special

diets are needed during pregnancy and disease. Adult dogs

should be fed according to their size and energy needs.

An adult dog needs at least 10% of its daily calories from

protein and a minimum of 5.5% from fats and it can contain

up to 50% carbohydrates, including 2.5% to 4.5% percent

fibre.Most adults should get two meals a day, although a

dog can eat just once daily. Giving two meals a day may

make it easier for the dog to digest the food and helps

control hunger.

Feeding Senior or Geriatric Dogs

Older dogs may not be as efficient in metabolizing dietary

protein as younger animals.They may actually require more

dietary protein than their younger counterparts to maintain

protein reserves and maximize protein turnover rates.

Somedogs begin old age considerably overweight, whereas

others may show some loss of condition.Feeding an

appropriate food with a different nutrient profile with

respect to energy, fat, or fiber content (increased or

decreased) may be needed to maintain optimal body weight

and condition. Geriatric dogsshould be monitored in a

preventive health program that includes periodic

assessments of body weight and condition. The

incidence of chronic degenerative organ disease increases

with age, and early diagnosis fosters earlier treatment and

more effective nutritional management.

Self-Feeding of Dogs

Self-feeding is the practice of allowing dogs unlimited

access to food. It is a practical and efficient means of

feeding the kennelled dog. Most notable among the many

advantages are:

• Each dog regulates its own food intake.

• Dogs are generally more content and much quieter.

• Less aggressive dogs do not have to compete for

food since they can eat when the others have finished.

• Dogs generally eat less at a feeding, but they eat

more often thus using their food more efficiently.

Caution must be taken with dogs that tend to overeat and

become obese. Self-feeding is not advisable for overweight

dogs. Some veterinary nutritionists do not recommend

self-feeding programs for puppies less than 4 months of

age.

For adult dogs:

• Put a continuous feeder and dry dog food in the

kennel.

• Continue regular feeding until the dog starts to eat

between meals and eats less of the regular food.

• Gradually reduce supplements such as meat, canned

food, scraps, etc.

• Gradually reduce the amount of water in the regular

meals so that finally regular meals consist solely of

dry dog food.

• Discontinue regular meals after the dog has adjusted

to dry food.

• An adequate supply of clean, fresh water should

always be available.

For Puppies:

• Offer gruel of dry food and water (about the

consistency of a milk shake) at 3 weeks of age.

• When the pups begin to eat the gruel, gradually reduce

the amount of water throughout the weaning period.

• After weaning, further reduce the water added to

the food until the pups are eating completely dry food.

• An adequate supply of clean, fresh water should

always be available.

References

1. Nutrition for the Adult Dog. Virginia-Maryland

regional college of veterinary medicine, Veterinary

Teaching Hospital, client information handout.

2. Merck Veterinary Manual.Tenth Edition

3. Lisa M. Freeman. Healthy Dogs, Web MD.

4. Nutrition - General Feeding Guidelines for Dogs. VCA

Animal Hospital.

U


HELMIMTHIASIS IN FISHU

Helminths are common in both wild and cultured fish. Fishfrequently serve as intermediate or transport hosts for larvalparasites of many animals, including humans. Helminths with

direct life cycles are most important in dense populations,and heavy parasite burdens are sometimes found. In general,heavy parasite burdens seem to be more common in fish

originating from wild sources.

Monogenean trematodes, which have direct life cycles, arecommon, highly pathogenic, obligatory parasites of the skinand gills. They are ~0.1-0.8 mm long and are best seen

microscopically. The worms can be identified by theircharacteristic hold-fast organ, the haptor, which is armed withlarge and small hooks. Aquarium and cultured fish are subject

to a rapid buildup of parasites by continuous infection andworm transfer to other fish in the tank or pond. Althoughmany species are host-specific, the more common types seen

in aquaria are less selective.

The 2 most common genera are Gyrodactylus and

Dactylogyrus . Gyrodactylus gives birth to live young, whichcan be seen within the body of the adult worm, and frequentlyare skin parasites; Dactylogyrus lays eggs and is principally

a parasite of the gills. Cleidodiscus is an important monogenefound on the gills of channel catfish. Neobenedenia andBenedenia are important monogeneans in marine fish. Infected

fish show hyperactivity and erratic swimming, often flashingabove the water surface or rubbing the sides of their bodiesagainst an object in the aquarium to dislodge the worms. Fish

become pale as colors fade. They breathe rapidly and distendtheir gill covers, exposing swollen, pale gills. Localized skinlesions appear with scattered hemorrhages and ulcerations.

Mortality may be high. To prevent the disease, introductionof infected fish should be avoided. Formalin is often thetreatment of choice for monogenean infestations. Multiple

treatments at weekly intervals are recommended forDactylogyrus because eggs may be resistant to chemicaltreatment. Organophosphates have been used successfully

in nonfood fish but are not approved for this use. Trichlorfon(active ingredient) as a prolonged bath (0.25 mg/L) is effective.Organophosphates break down rapidly as pH and temperature

rise, therefore slight increases in concentration may be

1Dr.Phaniraj.K.L M.V.Sc., Ph.D., 2 Kishore Kumar and Sushmita



necessary in marine systems. A bioassay is recommended ifpractitioners are uncertain how to proceed. Use oforganophosphates in ponds may be restricted by federal or

state environmental regulations. Monogenes on marine fishcan be removed using freshwater dips for 1-5 min, dependingon the tolerance of the species: however, eggs will not be

damaged or removed. Trichlorfon can be used in marinesystems, but some species are highly sensitive to it,particularly elasmobranchs. Increased ammonia levels should

be anticipated after chemical application. Praziquantel (2 mg/L) has become the treatment of choice for monogeneaninfestations of aquarium fish, particularly marine species. The

high cost of praziquantel is offset by excellent efficacy andtarget animal safety reported to date.

Digenean trematodes have complicated life cycles, withseveral larval stages that infect one or more hosts. With rare

exceptions, the first intermediate host is a mollusc, withoutwhich the life cycle generally cannot be completed. A diagnosisusually can be established by gross or microscopic

examinations that reveal the cercarial, metacercarial, or adultworms in any of the tissues or body cavities of the fish. Fishtend to form pigmented tissue encapsulations that encyst theparasites. Depending on the color of the cysts in the skin, the

condition is called black, white, or yellow grub disease.Heavily parasitized fish often are weak, thin, inactive, andfeed poorly. Treatment is not recommended.

Pond-reared, juvenile, tropical fish may develop severe gill

disease from metacercarial cysts in gill tissue. Although acutedeath is occasionally seen, infected fish more commonly dieduring harvest or shipping when they may be exposed to

suboptimal dissolved oxygen concentrations. Treatment ofinfected fish has not been successful; however, preventionof the disease by elimination of the intermediate host, a

freshwater snail, has been effective. Snails can be controlledin fish ponds by applying a copper sulfate treatment at nightwhen they are active. A molluscicide, Bayluscid®, is available

as a restricted-use pesticide in some areas (eg, Puerto Rico,Florida) for control of aquatic snails. It cannot be applied toponds containing live fish but is extremely effective in

eliminating snails if applied 1-2 wk before stocking.


Bolbophorus confusus is a digenean trematode that has

recently been reported to cause mortality in channel catfishfingerlings in production ponds in Mississippi, Louisiana,and Alabama. The definitive host of B confusus is the white

pelican, and the first intermediate host is the ram’s horn snail( Heliosoma spp ). Cercariae released from snails encyst infish tissue, forming metacercariae in any tissue, but the majority

are found in skin and skeletal muscle of the peduncle of juvenilechannel catfish. Severe disease occurs when metacercariaeencyst in visceral organs, particularly the posterior kidney

and liver. Involvement of these organs can result in apresentation similar to enteric septicemia or channel virusdisease, characterized by fluid accumulation in the abdomen

and exophthalmia. Skin and muscle lesions typically result inraised bumps that are white to reddish in color. Visceralinvolvement can result in high mortality (95%) of small fish.

Heavy infestation of older fish may result in anorexia, lethargy,and loss of condition. Digenea in skeletal muscle can result incondemnation of affected carcasses by processing plants.

Ponds at greatest risk to B confusus are those frequented bywhite pelicans. Pelicans are federally protected; however,

assistance for control of nuisance wildlife is available throughWildlife Services of the USDA. Snail control is an importantpart of an overall control strategy and requires a mix of chemical,

biological and aquatic plant control strategies. Copper sulfateis effective against snails but will not penetrate when they areburied in mud or sealed into their shells. Treatment is likely to

be most effective in summer and early fall when snails areactively feeding. Nocturnal application of copper sulfate hasbeen helpful in ornamental fish ponds, but care must be taken

not to precipitate an oxygen depletion by killing plants andalgae. Bayluscide® may be labeled in some states for controlof aquatic snails. Chemical control will not eliminate snails,

and efforts should be augmented by control of aquatic weeds.Snails climb emergent vegetation to lay eggs, so eliminatingvegetation can decrease reproduction. Finally, biologic control

may be attempted using black carp; however, these are anexotic species and stocking is prohibited in many geographicareas. Red-ear sunfish are also known to eat snails, but their

potential impact on snail populations has not been tested.Due to the complexity of this problem, and rapid generationof new information, practitioners are urged to consult with

extension and other aquaculture specialists.

Both larval and adult tapeworms are common in fish. Larvalforms encyst in visceral organs and muscle, while adultsusually are found in the intestinal tract. Aquatic Crustacea

are the most common intermediate host for fish; accordingly,

wild and cultured pond fish may be heavily infected.Diphyllobothrium latum , the broad fish tapeworm infectionof humans, is acquired by eating larval tapeworms in the flesh

of food fish. Aquarium fish may be purchased with heavycestode infections but have limited exposure once in theaquarium (unless fed infected intermediate hosts). There is

no safe, effective treatment for larval tapeworm infections.Corallobothrium spp are tapeworms occasionally found inthe intestinal tract of channel catfish; however, their clinical

significance is minimal. Larval migrations of the basstapeworm, Proteocephalus ambloplites , have beenassociated with reproductive failure in free-ranging

populations of largemouth bass. Although usually anincidental finding, heavy infestations of tapeworms have beenassociated with mechanical obstruction of the lumen of the

gut. The Asian tapeworm, Bothriocephalus acheilognathus

, is occasionally seen in carp and aquarium fish. It is usuallyfound in the anterior intestine and may be associated with

enteritis and degeneration of the intestinal wall. Praziquantelis the drug of choice for treatment of cestodes in fish, but it isnot approved for any aquatic use. It can be applied as a bath

(2 mg/L for prolonged immersion or 10 mg/L for 3 hr) or in amedicated food (50 mg/kg, 1 time).

Acanthocephala (thorny-headed worms) are common in wildfish as both larval tissue stages and adult intestinal parasites.They are more common in salmonid and marine fish.

Arthropods are the first intermediate host. Adultacanthocephala are easily recognized by their protrusibleproboscis, armed with many recurrent hooks.

Nematodes are common in wild fish that are exposed to the

intermediate hosts. Fish may be definitive hosts for adultnematodes, or they may act as transport or intermediate hostsfor larval nematode forms (anisakids, eustrongylids, and

others) that infect higher vertebrate predators, includinghumans. Encysted or free nematodes can be found in almostany tissue or body cavity of fish. Aquarium and cultured

pond fish may be heavily infected if crustacean intermediatehosts are present. Cyclops and Daphnia spp are commonintermediate hosts for Philometra sp , a nematode that is

pathogenic for guppies and other aquarium fish. These blood-red worms can be seen in the swollen abdominal cavity andprotruding from the anus of affected fish (red worm disease).

Capillaria spp are commonly found in aquaria fish, particularlyfreshwater angelfish. Heavy infections in juvenile angelfishhave been associated with poor growth rates and an inability


to withstand shipping and handling. Treatment with

fenbendazole (25 mg/kg for 3 days) is recommended, butefficacy has not been firmly established. Levamisole (10 mg/L) administered as a bath treatment for 3 days has also been

recommended. Ivermectin is highly toxic to aquarium fish,particularly cichlids, and its use is not recommended.

Leeches are parasitic bloodsuckers of fish and also serve asvectors for blood parasites of fish (eg, Trypanosoma ,Cryptobia , and haemogregarines). They can produce a

debilitating anemia due to chronic blood loss and disease.Leech infestations are most common in wild fish, but aquariumand pond infestations can occur by introduction of infested

fish, plants, etc. Trichlorfon (0.25-1 ppm in aquarium water,use higher dosages at higher pH) is effective but is notapproved for use in food fish, and environmental regulations

may restrict its use in outdoor ponds. Multiple treatmentsmay be required because eggs are resilient and juveniles maycontinue to hatch. Preventive measures include avoiding

leeches (ie, effective quarantine) and depopulating infestedaquarium fish. Infestations in recreational fishing ponds areoften self-limiting.

Copepods

Some copepods, during specific stages of their complicatedlife cycle, are obligatory parasites of finfish. They lose their

copepod form, including their appendages, and become rod-or sac-like structures specifically adapted for piercing, holding,feeding, and reproducing. Grossly, they appear as barb-like

attachments to the skin or gills, where they feed on blood andtissue fluids. They can cause hemorrhage, anemia, and tissuedestruction, as well as provide a portal of entry for other

pathogens. Many different species of these parasites can befound on freshwater and marine fish. The anchor worms,Laernea spp , are commonly found in a wide variety of

aquarium- and pond-reared fish, including goldfish and othercyprinids. Ergasilus spp infest the gills.

Lice ( Branchiuria ) are related to the parasitic copepods andhave flattened bodies adapted for rapid movement over the

skin surface. By means of hooks and suckers, they periodicallyattach for feeding by inserting the piercing mouth part (stylet)into the skin. Sea lice ( Lepeophtheirus salmonis ) are a

significant disease problem of pen-reared salmonids whichcan be treated with hydrogen peroxide. Argulus spp are licecommonly found on aquarium, pond-reared, and wild fish.

Trichlorfon at 0.25 ppm of aquarium water is the drug of choicefor treating infested aquarium fish but is not approved for usein food fish. Infested fish should not be introduced.



U


Homeopathic Approach to treat Bovine Clinical MastitisU

Bovine clinical mastitis is a disease of paramount

importance in female milch breeds. If not given attention

at proper time then not only it dampens the milk production

but also significantly affects the economy of the herd.

Most of the cases even after the treatment the animal

seldom attains its full productivity. The condition is also

worsened by the reduction in the size of the teat and the

udder. If any contaminants or debris get accessed inside

the udder then there is every possible chance of spreading

the infection to the other healthy compartments of the

udder. In female it sometimes affects other reproductive

parts (Metritis-Mastitis Complex). The prognosis in such

cases are always grave.

The medicinal approach (Allopathic approach) to treat such

anomaly is very costly and effective only to some extent

but the quality and taste of the milk cannot be restored. In

certain cases purulent discharge comes out from the udder

which if not treated properly then toxaemia prevails and

animal gradually succumbs to death. However homeopathic

treatment being cost effective and has no side effects on

animal health, now gaining momentum in animal practices.

Hence the farmer should consider the use of homeopathic

drugs given below to treat clinical mastitis with following

symptoms

• If the udder is swollen and hard and watery

or curdled milk comes out from it

Cal Sulph - 6×

Hyper Sulphur - 6×

Saraswat Sahoo1, Subhash Sharma2 and Netrapal Singh Sirohi3

Arawali Veterinary CollegeSikar, Rajasthan, India – 332001

1. Assistant Professor, Department of Veterinary Gynaecology and Obstetrics2. Assistant Professor, Department of Veterinary Parasitology

3. Prof and Head, Department of Veterinary Gynaecology and ObstetricsArawali Veterinary College, Sikar, Rajasthan, India -332001

Cal player – 1M

Phytollyca – 1M

Belladona – 1M

Administer three to four times orally daily

• If the udder and teat became cyanotic (Blue

colour)

Arsenicalbs – 200

Lychasis – 200

Agnus cast – 200

Administer 20 drops each, three to four times orally

daily

• If blood comes along with milk but there is

no inflammation or swelling of the udder

Arnica – 1M

Hypericum – 1M


• If milk present in the udder but flow of milk

from the teat is less

Conium Mac- 1M


• If there is change in the shape of the teats

Phytollyca – 1M

Pulsetilla – 1M


U


INTERNAL PROTOZOAN PARASITES OF FISHU

Sporozoans:Hexamita and Spironucleus spp are common, small (~9 µm),bilaterally symmetric, flagellated (4 pairs) protozoa mostfrequently found in the intestinal tract and occasionally inskin lesions or degenerating soft tissues of finfish. Thesegenera are similar but differ slightly in the position and shapeof their nuclei (2 within 1 organism). Pathogenicity of theseorganisms is variable and correlated with the number present.If there is a question as to whether treatment is warranted, thenumber of organisms present can be assessed in a wet mountof intestine. If <5 organisms per low-power field (LPF) arepresent, treatment is probably not necessary; if 5-15organisms/LPF are present, treatment should be administeredif fish are in poor condition; and if >15 organisms/LPF arepresent, treatment is strongly recommended. The onlytreatment available for hexamitiasis is metronidazole (use onlyin ornamental species), which should be given orally but canbe administered as a bath if fish are anorectic. The number oforganisms present in infested freshwater angelfish increasesdramatically after shipping and handling. Chronic problemshave been seen in fish maintained in unsanitary or crowdedconditions. Preventive treatment of ornamental cichlids isrecommended before shipping, and broodstock should beevaluated periodically. The hatchability of eggs from heavilyinfested adult angelfish (freshwater) seems to be significantlydecreased; resultant fry may be weak with poor longtermsurvival.Cryptobia and Trypanosoma spp are slender, elongated (6-20 µm), actively motile, biflagellated protozoa that are easilydetected in fresh blood and tissue smears of both marine andfreshwater finfish. Hematogenous forms are generallydescribed as Trypanosoma and have a well-developedundulating membrane. Trypanosomes may be transmitted byleeches and have been associated with anemia in blue-eyedplecostomus imported from South America. Cryptobiaiubilans has been associated with granulomatous disease inAfrican cichlids and discus.Clinical disease is manifest by severe weight loss and cachexia.Clinically affected fish should be culled. Presumptive diagnosiscan be made from microscopic examination of fresh tissue.Typically, granulomas will be found in the stomach, whichmay be visibly thickened. Acid-fast material will not be foundin granulomas caused by Cryptobia . Motile flagellates maybe visible using magnification of 400¥ or greater. Transmissionelectron micrographs are required to confirm the diagnosis ofC iubilans.

1Dr.Phaniraj.K.L M.V.Sc., Ph.D., 2 Kishorekumar and Sushmita



Coccidiosis, while common in freshwater or marine finfish, israrely diagnosed in live fish. Many species of finfish areaffected. The life cycles of many fish coccidia are unknown,and some involve >1 host to complete their development. Inaddition to intestinal infection, the internal organs also arecommonly affected; sporulated Eimeria -like oocysts andsexual and asexual stages are commonly found in direct smearsand histologic sections of the internal organs. Sulfamethazine,at 22-24 g/100 kg of fish wt/day in the feed for 50 days at 50°F(10°C), is used to treat food fish (21-day withdrawal time) insome countries. For aquarium fish, 10 ppm in the aquariumwater once a week for 2-3 wk has been reported to bepreventive, but safety and efficacy data are sparse.Myxosporidians are common fish parasites. Themyxosporidian spore consists of 2 valves, a suture line, and1-4 polar capsules that contain coiled, extensible filamentsand an infective central body called the sporoplasm. Evidencesuggests that myxosporidia have indirect life cycles and useother aquatic organisms (eg, annelids) as intermediate hosts.Hence, myxosporidian infections are more common in, andmore pathogenic for, wild fish or fish reared intensively inoutdoor fish ponds. The organisms tend to be host- and tissue-specific. Accordingly, expression of the disease is related tothe specific pathogen and host.Myxosoma cerebralis , an important pathogen of youngsalmonids, is responsible for whirling disease, also known as“blacktail.” Typically, infected fingerlings show rapid tail-chasing behavior when startled, and the peduncle and tailmay darken significantly. As infected fish age, skeletaldeformity may result from damage to the cartilaginousstructures, particularly the skull and vertebral column.Recovered fish remain carriers, and adults do not show signs,although skeletal deformities do not resolve. The disease canbe prevented by purchasing uninfected breeding stock andmaintaining them in an environment free of the intermediatehosts. A presumptive diagnosis of whirling disease is madeby detection of spores from skulls of infected fish. Samplescan be submitted to a fish disease laboratory, or proceduresdescribed by the American Fisheries Society can be followed.Diagnosis may be confirmed histologically or serologically.Whirling disease is of regulatory concern in some states.Salmonid ceratomyxosis is caused by Ceratomyxa shasta , amyxosporidian endemic to specific watersheds in the Pacificnorthwest. The disease occurs in wild fish as well as in fishfrom hatcheries that use contaminated water. The most typicalpresentation includes hemorrhage and fibrinous inflammation


in the posterior intestine, but other visceral organs andmusculature can also be infected. Grossly, fish may appearemaciated, with a distended abdomen and hemorrhagic vent.A presumptive diagnosis can be made by examination of awet mount of the posterior intestine and visualization of thekidney-bean-shaped trophozoites. The presence of theorganism can be confirmed histologically. Some statesconsider C shasta a reportable disease.Proliferative gill disease of catfish is caused by themyxosporidian Aurantiactinomyxon ictaluri . The organismhas a complex life cycle, with the oligochete worm Dero digitataserving as the intermediate host. Channel catfish may be anaberrant host for A ictaluri , and the disease is usuallyassociated with new ponds or previously infected ponds thathave been drained and refilled. Although proliferative gilldisease can cause catastrophic mortality approaching 100%,losses may be as low as 1%. Disease occurs at watertemperatures of 16-26°C, and mortality is exacerbated by poorwater quality, particularly low dissolved oxygen or high levelsof un-ionized ammonia. Gills of affected fish are severelyswollen and bloody, resulting in the colloquial name“hamburger gill disease.” A presumptive diagnosis can bemade from a wet mount of infected gill tissue, in which filamentsappear swollen, clubbed, and broken. Cartilaginous necrosisis strongly supportive of a diagnosis of proliferative gilldisease; however, histology is required for confirmation.Many species of myxosporidians produce nodular or cysticlesions in the skin, gills, muscle, or visceral organs of fish,depending on their host species and tissue preference.Henneguya is commonly found in white, cystic skin lesionsof cultured channel catfish and aquarium fish; it is easilyidentified by the forked-tail appendage of the spore seenmicroscopically. If ponds are dried and limed heavily, infectioncan be eliminated, apparently by reduction of the intermediatehosts. Aquarium infection can be self-limiting in the absenceof intermediate hosts. Henneguya may also be seen in wetmounts of gill tissue. Although an occasional cyst may beconsidered an incidental finding, severe damage has beenassociated with diffuse distribution of interlamellar cysts.Renal dropsy in pond-reared goldfish is caused by themyxosporidian Sphaerospora auratus . The disease ischaracterized by renal degeneration and ascites and is usuallydiagnosed by identification of spores in histologic sectionsof the kidney. Newly purchased pond-reared goldfish placedin aquaria may show signs of the disease, including death.No practical treatment is available. The carp-dropsy complexis a disease of carp and goldfish characterized by dropsy andexophthalmos. It is associated with S angulata infection andmay be complicated by viral infections (such as spring viremiaof carp), carp swim-bladder disease, or bacterial septicemias.Deaths may be acute or occur over a 6-mo period. The responseto drug treatment is generally poor.Proliferative kidney disease (PKD) is one of the mosteconomically important diseases affecting salmonid industries

of North America and Europe. Rainbow trout are particularlysensitive to the disease, although all salmonids seemsusceptible. PKD is caused by an unnamed myxosporidianparasite, sometimes referred to as the PKD parasite. PKD hasbeen reported in both captive and free-ranging salmonidpopulations. It occurs most commonly in the summer whenwater temperatures are >12°C, and the parasite primarily infectsyearling and younger fish. Clinical signs include lethargy,darkening, and fluid accumulation indicated by exophthalmos,ascites, and lateral body swelling. Infected fish are frequentlyanemic, resulting in gill pallor. Grossly, the posterior kidneyappears gray, mottled, and significantly enlarged. Presumptivediagnosis can be based on observation of suspect organisms,10-20 µm in diameter, in Giemsa-stained wet mounts of kidneytissue. Histologic examination of infected tissue, stained withH&E, is required for confirmation. Avoidance is the bestpreventive measure, although losses may be minimized byimproved husbandry. Bacterial infections, particularlyAeromonas salmonicida , are common sequelae of PKDepizootics and, if uncontrolled, can result in substantiallyincreased mortality. There is no treatment; however, fish thatrecover from the infection are resistant to subsequentoutbreaks. Infected stocks in nonendemic areas should bedepopulated, the premises sanitized, and disease-free stockobtained for replacement.Microsporidians are tiny, intracellular, spore-formingorganisms with single polar filaments that are commonparasites of finfish. They are host- and tissue-specific andcan also infect helminth parasites of fish. The spores areextremely resistant.Pleistophora ovariae infects ovarian tissue of golden shiners(bait fish), resulting in sterility. It is an important disease inthe bait-fish industry. The organism has no intermediate hostand is transmitted horizontally (through ingestion of infectivespores) or vertically (through infected ova). Fertility declinesas fish age, eventually resulting in sterility. Grossly, infectedovarian tissue appears marbled. The diagnosis is confirmedby examination of a wet mount of suspect tissue, revealingthe presence of microsporidian spores. Although there is notreatment, the problem can be managed by discarding femalebroodstock when they reach 1 yr of age and replacing themannually. Although the young females remain infected, thedisease is not yet sufficiently advanced to have a major impacton fertility.Neon tetra disease is caused by Pleistophorahyphessobryconis , which infects the skeletal musculature ofa number of species of aquarium fish, including tetras,angelfish, rasporas, and barbs. Infected fish may exhibitabnormal locomotion caused by muscle damage, and muscletissue may appear marbled or necrotic at necropsy. The parasiticspores are readily visualized in wet mounts of infected tissue.There is no treatment for the infection, although removal ofmoribund fish helps prevent transmission by eliminatingcannibalism within the population.

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MAMMARY GLAND DEVELOPMENTDURING DIFFERENT GROWTH STAGES IN CATTLE

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The mammary gland is the milk secreting structure, which

includes a teat, a duct system and

lobes (lobules) of secretory tissue drained by the duct

system. Mammary glands are modified

sweat (sudoriferous) glands, which secretes milk (exocrine

gland) and serve as accessory glands

to the reproductive system. Mammary development begins

when the animal is an early fetus and proceeds beyond

initiation of lactation. The mammary gland is one of a few

tissues in mammals, which can repeatedly undergo growth,

functional differentiation, and regression.

Mammary gland development during the fetal period

Mammary development before birth: the early embryo

has three distinguishable layers of cells which ultimately

give rise to various tissues and organs of the body. These

are: the ectoderm (outer layer) which gives rise to the

skin (epidermis) and nervous system; the mesoderm

(middle layer) which gives rise to muscle, blood vascular

system and sex organs; and the endoderm (inner layer)

which gives rise to the alimentary canal and digestive

glands. The mammary gland is derived from the ectoderm

and mesoderm layers.

Mammary band: The mammary band is found at ~ 32

days in the bovine embryo (~1cm long). The mammary

band persists for about 1 week. The mammary band is a

broad band of ectodermal cells running on either side of

the trunk from the upper limb to the lower limb.

Arvind Ku. Pandey1, Saraswat Sahoo2 and Deepak Ku. Kashyap3

Arawali Veterinary College, Sikar, Rajasthan, India -332001

*Corresponding Author E-mail address- [email protected]

1. Assistant Professor, Department of Veterinary Physiology and Biochemistry,

2. Assistant Professor, Department of Veterinary Gynaecology and Obstetrics,

3. Assistant Professor, Department of Veterinary Surgery and Radiology,

Arawali Veterinary College, Sikar, Rajasthan - 332001

Mammary streak: there is further development of the

mammary gland.

Mammary lines: The mammary streak becomes further

differentiated to form the mammary line by the 4-5th week

of fetal age when the bovine embryo is about 1.4-1.7 cm.

the mammary line is a narrow ridge of slightly taller

ectodermal cells. Resting on a strip of condensed

mesenchymal cells (cells from the mesoderm).

Mammary crest: The mammary lines begin to shorten

and the ectodermal cells begin to divide and grow into the

mesenchymal cell layer.

Mammary hillock: The ectodermal cells continue an

inward growth into the mesenchymal layer. In cross-

section, the mammary hillock appears as a dome of

ectodermal cells growing into the mesenchymal cell layer.

Mammary bud: The ectodermal cells continue to grow

into the mesenchymal layer, resulting in formation of a

spherical or globular structure. The mammary bud is

formed early in the second month in the bovine when the

embryo is about 2.1 cm long.

The mammary bud formation is a critical stage in mammary

development. It is after the bud forms when several other

important processes begin. The ectodermal layers sink in

the mesenchyme leaving forming a dimple on the embryo’s

surface (mammary pit). The mammarybud stage marks

the beginning of differentiation patterns which distinguish

various species. The mammary bud stage also marks the

point at which glands of females and males can be

distinguished.


Age of embryo (d) Crown-rump length Stages of development in bovine

(mm) embryo mammary gland

32 14

34 16

35 17

37 19

40 21

43 25

Early teat development:

In the bovine the early origins of the teat development

begin by about 60 days when the fetus is about 8cm long.

Rapid growth of the mesenchyme around the mammary

bud raises the area containing the bud up from the

surrounding surface. Blood vessels begin to form in the

mesenchymal area associated with the bud. Also, an

invagination of the mammary bud cells into the

mesenchyme occurs, pushing the mesenchymal cells aside

. this becomes the primary sprout by 80 days. The primary

sprout ultimately gives rise to the gland cistern, but at this

early it is still a solid core of cells.

Secondary sprouts:

in the bovine fetus, the secondary sprouts branch from

the primary sprout at about 13-14 wks, just prior to

canalization of the primary sprout. Secondary sprouts are

still a solid core of cells at this time. The secondary sprouts

will form a major ducts leading to the major lobes of the

gland.

Canalization:

the process of forming a lumen in the solid core of epithelial

cells in the primary and secondary sprouts is called

canalization and begins at about 100 days in the bovine

fetus (about 19cm long).


Primary and secondary sprout formation in embryonic mammary gland

Canalization in embryonic mammary gland


Mammary gland development during the prepubertal

period

Birth to puberty: mammary growth in the bovine is

isometric (grows at the same rate as general body growth)

for the first 2-3 months after birth. The duct system

enlarges a little. The increase in udder size results from

the continued increases in fat pad and connective tissue.

No development of secretory alveoli occurs at this time.

At about 2-3 months after birth, the allometric growth

begins (growth rate faster than the rest of the body). In

the calf, this includes extensive growth and development

of the duct network which invades the surrounding adipose

tissue(fat pad). No alveoli are formed. During pregnancy

the ducts will differentiate into milk secretory cells.

Therefore the formation of a duct network that occurs

until puberty will determine the extent of lobulo-alveolar

development during gestation. The allometric growth phase

lasts until about 1 yr of age, when the mammary growth

rate returns to isometric growth. The allometric growth

will not occur in the absence of fat pad. These apparently

are local interactions between the fat pad and the growing

duct system, through the presence of cytokines And

growth factors, but these interactions are not fully

understood yet.

Influence of growth hormone and leptin on mammary

development: daily injection of somatotropin (growth

hormone) to heifers from 8-15.6 months of age resulted

in increased mammary parenchyma and decreased

extraparenchymal tisuue as compared to controls. Growth

hormone stimulates mammary growth through increasing

the hepatic synthesis of insulin-like growth factor-1 (IGF-

1), which is a potent mitogen for mammary cells.

Another protein that influences mammary development

indirectly and that could explain how excessive fattening

could cause mammary impairment in leptin, which is

produced by adipocytes and decreases IGF-1 induced

bovine mammary cell proliferation.

Mammary gland development during the post pubertal

period

Puberty to conception: at the actively growing end of

the ducts, where the outermost limits of ductal elongation

invade the fat pad are actively growing structures called

Terminal ductile lobular units (TDLU). Terminal ductile

lobular units represent the structures where elongation and

branching of the ducts is occurring and estrogen stimulated

cell division is occurring.. in general, estrogen causes cell

multiplication at the tip of the TDLU and enlargement of

the ducts (lengthening and branching of ducts), while

progesterone causes duct and ductile cells to multiply,

leading to ductile development and duct enlargement or

widening.

Synergy between estrogen and progesterone is observed

during pregnancy when both hormones are present in high

concentration. Elevated blood concentrations of estrogen

and progesterone together establish the conditions

necessary for the exponential growth which occurs during

pregnancy.

Prolactin is often associated with initiation of lactation and

galactopoeisis, but also has mammogenic effects. Prolactin

receptors are present in the fat pad of some species as

well in the epithelium. Prolactin may act on both epithelium

and and stromal components of the growing mammary

tissue.

Growth hormone (somatotropin) administration to cattle

is known to stimulate milk production during lactation.

This effect is indirect in that growth hormone stimulate

secretion of insulin-like growth factor-1 (IGF-1) from the

liver, which in turn mediates many of the galactopoeitic

effects of growth hormone during lactation. Growth

hormone also acts as a mammogenic hormone and can

stimulate mammary growth at various stages of

development. Mammary expression of IGF-1 is regulated

by growth hormone, estrogen and positive feedback

stimulation from proliferating epithelial cells.

Placental lactogens are secreted from the placenta and

may have prolactin or growth hormone like activities,

depending upon the species.

Other hormones are also required for mammary growth.

Including glucocorticoids, thyroid hormones and insulin.


Hormonal regulation of mammary developmentduring pregnancy:Estrogen and progesterone: Optimal mammary growthrequires both estrogen and progesterone. Duringpregnancy the mammary tissue has estrogen receptorsand progesterone receptors. During lactation the mammarygland has estrogen receptors, but not progesteronereceptors. Concurrently elevated estrogen andprogesterone, such as during pregnancy establish theconditions necessary for genomic cell multiplication tooccur. For example from one original cell, 8 cell divisionswill yield 128 cells. Concurrently elevated estrogen andprogesterone also results in lobulo-alveolar growth, whichis the characteristic of the type of mammary tissuedevelopment that occurs during pregnancy. In the cow,progesterone is elevated throughout gestation (requiredfor maintenance of pregnancy), while estrogen is elevatedduring second half of gestation. Consequently, most ofthe mammary growth during first half of gestation is mainlyductal growth and lobule formation. In the second half ofgestation, ductal growth continues, but most growth islobulo-alveolar.

Insulin: Required for maintaining mammary tissuefunction in vitro. Mammary cells are resistant to insulinbefore conception, but they become sensitive to insulinduring gestation and lactation, and become insensitiveduring involution.

Thyroid hormones: thyroid hormones are involved in theoverall metabolic rate and oxygen consumption of thebody.

Mammary gland development during lactation:A major aspect of the mammary development in thelactation period is that the progesterone source (corpusluteum) is lost, so their receptors cease to exist and onlyestrogen is available for mammary development. Thenumbers of mammary cells in the lacatating mammarygland are critical for milk production. Mammary cellnumbers continue to increase even after parturition.Mammary wet weights and total DNA continue to increasein early lactation. The impact of this increased mass ofmammary tissue on milk production can be substantial insome species. For example, total DNA content inmammary gland of rats is highly correlated (rsquared=0.85) with litter weight gain. In cows, mammaryDNA increased by 65% from 10 days pre-partum to 10days post-partum. Cell numbers in the cow mammarygland have not been determined throughout the lactatingperiod.

Cellular changes during Mammary gland developmentafter birthAutocrine and paracrine regulation of mammarygrowth: Autocrine and Paracrine factors (local growthfactors) play a major role in mammary growth. Many ofthe effects of the steroid hormones on mammary growthare mediated by local growth factors at the mammarytissue level. These include an interaction between thedeveloping mammary epithelial structures and themammary fat pad. Along with IGF, number of othergrowth factors have positive or negative effects onmammary gland development. Local production oftransforming growth factor –â (TGF-â) inhibits mammarygrowth, such as during the pre-pubertal period andbetween the estrous cycles. Epidermal growth factor)EGF) and transforming growth factor –á (TGF-á)produced in the mammary tissue stimulate the mammarycell proliferation.Mammary gland development during pregnancy:Mammary growth (of the mother) accelerates duringpregnancy. This is fastest during later stages of pregnancy,which coincides with the most rapid period of cell growth.Pregnancy is often considered to be the period of mostextensive mammary growth. Extensive lobulo-alveolardevelopment occurs only during pregnancy. The milksecretory cells develop only during pregnancy, thereforethis period is extremely important in determining thenumber of secretory cells in the in the lactating gland andthe subsequent production of milk. Correlations betweenthe total DNA in the lactating gland and milk yield forvarious species range between 0.5-0.85. the correlationbetween the number of mammary cells estimated in thelactating gland of rats and milk yield is 0.85, indicating aclose relationship between the cell number of lactatinggland and milk yield.

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Preservation of wild animal cadaver and samples forits use as model teaching tool and management

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Dead body is best material to learn the technique beforeworking on live animal, which allows experts to minimize

potential damage and rehearse before trying it real. For thispreservation of dead body is essential as they are liable todesiccate or putrefy by bacteria and fungus. Preservation of

wild animal is very important for understanding their anatomy,physiology and behavior. As behavior based approach toensure conservation fitness and welfare is need of the time

for wild life. Carcasses or residue of dead animals, scats,feathers and other biological material found in the wild maybe very useful for obtaining data about wild animals with little

disturbance of live animals or their habitat.

Common material used in mummification in ancient time in

Egypt (4000BC) is linen, saw dust, lichen, bees wax, resins,onion, nile mud, linen pads, frankincense etc. Mummificationis removal of body fluids followed by wrapping the dead body

in linens. Until 19th century the dead bodies were preservedby using very toxic chemical, Arsenic. It was replaced byformaldehyde after its discovery in 1867. Preservation of dead

body using formaldehyde (toxic and carcinogenic) makes thebody stiff and fragile, and was not suitable for understandinghow organ will respond to a particular surgical procedure.

During 21st century the methods of body preservation includeplastination, silicon S 10 procedure, Cor Tech Roomtemperature procedure and Epoxy E 12 procedure, Polyester

P 35 procedure etc. These methods are advance version ofplastination.

Types of specimens used for preservation (Nagorsen andPeterson, 1980):

1. Entire fluid preserved animals: For studying

anatomy and histology

2. Skin with accompanied organs like skull: Forstudying bones, skin, hair quality and molting

pattern

S. Chaurasia1, R. Menaka2 and T. K. S. Rao3

Department of Veterinary AnatomyVanbandhu College of Veterinary Science and Animal Husbandry,

Navsari Agricultural University, Navsari 396 450 Gujarat1, 2, 3 Assistant Professor, Vanbandhu College of Veterinary Science and Animal Husbandry,

Navsari Agricultural University,396 450 Navsari Gujarat, India Email: [email protected]

3. Mounted skin with partial or complete skeleton orfreeze-dried specimens

4. Entire skeleton for study of anatomy, age

determination, geographic variataion, sexdetermination etc.

Preservation of specimens in the field: In the field condition,

there may be limited access to material and equipmentnecessary, therefore basic preservation with more simplemethod is essential before final preparation as permanent

collection:

Short term preservation of whole animals: In cold to moderate

climate without refrigeration the animals may be stored inshade for 4-5 hours, after this period viscera begin todecompose (Hangay and Dingley, 1985).

Formalin preservation: After weighing and biometry of animal

label is tagged. Small specimens up to 100 g can be fixedwhole by submerging them in 10 percent buffered formalin. Incase of large sample size or whole animal, body cavity can be

filled with formalin solution by injection until carcass becometurgid and firm. The ratio of formalin to carcass must be atleast 12:1 to assure good fixation. Formalin hardens the

specimen, discolor the fur, soften the bones and preventmicrobiological examination therefore it is better to preservein alcohol especially for long term preservation (Munson,

2000; Rabinowitz et al., 2000).

Preservation in alcohol: 70-90 percent alcohol is generallyused for preservation of carcass. The carcass is well preservedif intestine is removed prior to storage.

Preservation by cooling and freezing: It is recommended thatbefore freezing or cooling fur should be removed for quick

cooling of carcass. However freezing reduces the quality oftissue therefore histological and pathological examination isdifficult in this method (Wobeser and Spraker, 1980).


The preservation is also done by embalming. Embalming is a

technique to prevent a dead body from decaying by treatingit with special substance to preserve it. The objective ofembalming is to keep cadavers fresh and withstand handling

for long time.

Common embalming procedure: Arterial embalming with a

gravity tank apparatus fixed five feet above the dead body orby using pressure pump directly.

Preparation of embalming fluids: 2.5 liters of 10 percentformalin, 1 liter common industrial spirit, 1 liter glycerine and

500 ml liquid phenol. 5 liters per dead body is sufficient forpreservation. Here formalin is used as fixative. It preservesterilize and hardens the tissues. Glycerin softens the tissue

and keeps the muscle moist. Phenol prevents fungal infection.Water keep tissue wet and prevents them from drying up.

• Simple 10 percent formalin alone is in use asembalming agent in many veterinary and medical

colleges.

Embalming techniques: Embalming utilizes the artery for

injection of embalming fluids so called “Arterial embalming”there by utilizing whole vascular system.

Injection points: Two common points for injection is utilizedfemoral and common carotid artery.

Thiel method (Soft embalming): After formaldehyde,embalming fluid of salt, antiseptic like boric acid, ethylene

glycol and very small quantity of formaldehyde in combinationwas tried with good result. Around twenty liters ofpreservative fluid required per dead body of adult to be

preserved. The method is unique as the cadaver preservedby this method show no detectable odour, flexibility in bodyparts like living. The colour of organ also preserved with

antimicrobial ability. Soft embalming also helps in excellentvisualization of Anatomical structure via ultrasonography(McLeod et al., 2013). Ultrasound image of Thiel embalmed

cadaver were good and matched the quality seen in patient.Nerve easily identified and tracked, fluid visualized easily nearnerves. Anesthetist can use this technique to study regional

block by anesthesia using ultrasound imaging.

• Larssen solution is used in veterinary cadaverpreservation which provides acceptable cadaverquality and tissue handling for its use in surgical

instruction and teaching.

• Composition of Larssen solution: Sodium chloride500g, Sodium bicarbonate 900 g, Choloral hydrate

1000 g, sodium sulphate 1100 g, Formalin 10

percent solution 500 ml and 1 L distilled water.

Use of Honey and Vinegar as embalming agent: The honey is

a sweet aromatic viscid liquid derived from nectar of plantsand modified by honey bee. The honey and vinegar can bemixed in equal proportion and used for embalming at the rate

of 5 liters per dead body. Embalmed body then placed in bigformalin jar, it will remain fresh for 6 months for dissection andstudy.

Ethical source of cadaver: Animal cadaver and tissue obtained

from animals that have died naturally or in accidents, oreuthanized to natural un-curable disease or non recoverableinjury. A cadaver or tissue is used which is destined for disposal

(Martinsen and Jukes, 2007).

For osteological specimen: Maceration by boiling in hot waterwith soap for 1 hrs and then cooled at room temperature anddrying followed by bleaching using 30 percent hydrogen

peroxide.

For skin preservation with hair: Skin can be dried in sun or

using fire, stretched between pegs. Salting, powdered boraxor cold ash could further preserve it. Color change of hair iscommon in preservation especially with formalin.

Hair: Hair may have microscopical features allowing

identification and characterization of animals especially thesympatric species. Hair may be collected from scats of wildanimals for its identification. Hair sample can also be used for

DNA isolation identification and analysis.

Samples for food analysis: If a carcass is found in the wild,

collection of content of stomach only is not sufficient thesample should be collected from entire alimentary tract forfood analysis. As some of the food items retain in stomach for

few minutes. The content of tract can be preserved in 5%formalin or 30-40% alcohol (Rabinowitz et al., 2000)

Nutritional analysis from feces or content of gastrointestinal(GI) tract: Nutritional analysis and slide preparation require

more time therefore preservation of material in the field andfurther analysis in lab is better option.

Animals preserved as puppet: Puppet is used for teachingmodel using glycerine.

Plastination: Plastination has revolutionized the approach ofpresentation, human and veterinary anatomy to the students.

Study suggests plastination technique is innovative teachingand learning system (Latorre et al., 2007). Term plastinationderived from Greek word plassein meaning to shape or form.


Plastination is used for wet biological specimen preservation.

Plastination is technique of tissue preservation developedby Gunther Von Hagen (1978) of Germany. In this technique,all water and most all of lipid in biological tissues are replaced

by curable polymers (silicon, epoxy, polyester) which aresubsequently hardened by resulting in odorless dryspecimens. Silicon is used for whole specimen, thick body

and organ slice. Epoxy is used for thin, transparent body andorgan slice. Polyester polymer used for brain slice todifferentiate grey and white matter clearly. It is very good

model for teaching especially the neuro-anatomy.

Plastination techniques: The technique of plastination

consists of four steps: 1. Fixation, 2. Dehydration and de-fatting using acetone, 3. Forced impregnation: impregnationof polymer replacing acetone and 4. Hardening or curing using

gaseous hardner like silicon or by UV light and heat (polyester,epoxy) (Dhingra et al., 2006).

• Shellac (natural substance used in making varnishto protect surface and make them hard) is a non

toxic preservative for embalming in human cadaver.The shellac show protective properties by forminga superficial protective film.

Conclusion:

Preservation of cadaver started from mummification inEgyptian era to modern plastination techniques with some

interventions. Preservation of cadaver especially by meansof plastination helps even untrained people can look intoform and structure of body in new way. It helps the students

to know the site of different operation. Different muscles bloodvessels and nerves around the site and best approach towardsit. Plastination provides artistic look to the scientific specimen

for understanding anatomy easily. It must be used to enhancethe quality of education. Preservation of cadaver and samplesfrom wild animal is very essential to understand anatomy

physiology and behavior which will strengthen theconservation strategy especially for threatened species.Samples collected from wild can be used for census, microbial

and parasitic analysis. Study of histo-pathology of etiologyfrom preserved samples from suffering animals may furtherreduce the incidence by intervention in-vivo.

References:

1. Dhingra, R., Taranikanti, V. and Kumar, R. 2006.Plastination: teaching aids in anatomy revisited. Natl

Med J India. 19 (3): 171.

2. Hangay, G. and Dingley, M. 1985. Biological MuseumMethods. Vertebrates. Academic Press, New York, l:xv+ 1-379.

3. Latorre, R. M., Gracia-Sanz, M. P., Moreno, M.,Hernandez, F., Gil, F., Lopez, O., Ayala, M. D., Ramirez,G., Vazquez, J. M., Arencibia, A. and Henry, R.W. 2007.

How useful is plastination in learning anatomy?,Journal of Veterinary Medicine Education., 34(2):172-6.

4. Martinsen, S. and Jukes, N. 2007. Ethical sourced animalcadavers and tissue: Consideration for education andtraining, Proc. 6th World Congress on Alternative andAnimal use in Life Sciences. August 21-25, Tokyo, Japan;AATEX 14, Special issue 265-268.

5. Mc Leod, G., Eisma, R., Schwab, A., Corner, G., Soames,

R. and Cochran, S. 2013. An evaluation of Theil-emblamed cadavers for ultrasound based regionalanesthesia training research. Ultrasound, 18:125.

DOI:10.1258/ult.2010.010016.

6. Munson, L. 2000. Necropsy procedures for wild animals.With input from: W. B. Karesh, M. F. McEntee, L. J.Lowenstine, M. E. Roelke-Parker, E. Williams and M. H.Woodford; Illustrations by D. Haines). Pp. 203-224 in:

Conservation research in the African rain forests: atechnical handbook. White, Lee; Edwards, Anne (eds.),Wildlife Conservation Society, New York. ISBN: 0-

9632064-4-3 (english), ISBN: 0-9632064-5-1 (french).

7. Nagorsen, D. W. and Peterson, R. L. 1980. Mammal

Collectors’ Manual: a Guide for Collecting,Documenting, and Preparing Mammal Specimens forScientific Research, Life Sciences MiscellaneousPublications, Royal Ontario Museum, Toronto, Canada,79 pp.

8. Rabinowitz, A. Hart, J. and White, L. 2000: Informationfrom dead animals and their curation. Pp. 191-201 in:

Conservation research in the African rain forests: atechnical handbook. White, Lee; Edwards, Anne (eds.),Wildlife Conservation Society, New York. ISBN: 0-

9632064-4-3 (english), ISBN: 0-9632064-5-1 (french).

9. Wobeser, G. A. and Spraker, T. R. 1980. Post-mortemexamination. Pp. 89-98 in: Wildlife ManagementTechniques Manual, 4th edition, S. D. Schemnitz (ed.),

The Wildlife Society, Washington, D.C. ISBN: 0-933564-08-2.

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Role of Micronutrients in Animal ImmunityU

Nutrition of animal interacts with their immune system.Major nutrients like energy, protein, fat and micronutrientslike vitamins and minerals are play a vital role in evoking animalimmune response. The relationship between nutrition anddisease resistance is complex but it is well documented thatthe micronutrients play important role in animal immunity.Minerals like zinc, copper chromium, iron, cobalt, seleniumand manganese and vitamins like Vitamin E, Carotenoids (betacarotene) and vitamin A and vitamin C are having significantrole in animal immune status. Cattle can have sufficient vitaminand minerals intake for adequate growth and reproductiveperformance but not have optimal immune response.

Stress associate with weaning and transportation has anegative effect the immune system. This stress typicallyoccurs when the animal is exposed to a variety of infectiousagents as a result of marketing /transporting/ managementprocedures. Nutrition can interact with these two primaryfactors mostly likely us a result of infectious agents.Preweaning nutritional deficiencies or through decreases feedintake associated with stress. Decreased feed intake/ nutrienthas further depressed the immune function and potentiallyincreases susceptibility to infection. Nutrients derived fromdietary proteins, carbohydrate and fats as well asmicronutrients, vitamins and minerals interact with immunecells systematically in the circulating blood, regional lymphnodes and specialized immune system of gastrointestinal tract.

Immunology:Immunity refers to reactions by an animal’s body to foreignsubstances such as microbes and various macromolecules,independent of a physiological or pathological result ofreaction. Immunity is generally classified as either innateimmunity (natural) or acquired (specific). Innate immunityincludes physical /chemical barriers, the complement system,phagocytes such as macrophages, neutrophils, and naturalkiller cells and macrophages derived cytokines such as alphaand beta interferon’s and tumor necrosis factor. Acquiredimmunity, which is induces by natural exposure or vaccination,includes antibiotics, lymphocytes and lymphocyte-derivedcytokines such as interleukins and transforming growth factor.

Acquired immunity if further divided into either Humoral orcell mediated immunity. Humoral immunity is mediated by B-lymphocytes, which respond to antigens to become antibodyproducing cells and memory cells and provide defense againstintracellular microbial infection. In cell-mediated immunity, the

Raju Kushwaha, Muneendra Kumar, Vinod Kumar, Debashis Roy, Shalini VaswaniDepartment of Animal Nutrition,

College of Veterinary Science and Animal Husbandry,DUVASU, Mathura-281001

T lymphocytes and associated cytokines provide defenseagainst intracellular pathogens and tumor cells. Humoralimmune response can be measured by estimating the antibodyproduction by zinc turbidity method.

Zinc Zinc is an essential trace element for the immune system.The innate as well as specific parts of immune system areinfluenced by Zn. Zinc is component of numerous enzymeslike Superoxide dismutase (SOD), RNA polymerase, DNApolymerase, Thymidine kinase and Ribonuclease. Zincdeficiency results in atrophy of the thymus and increaseleukocyte count with reduced number of lymphocytes.Immature neutrophils are elevated in zinc deficient animals.Zinc is important in activation of B cells and NK cells. Zinc isessential cofactor for the thymic hormone thymulin. Thymulinis secreted by thymic epithelial cells and induces differentiationin immature T cells. Zinc influences host defense mechanismvia: phagocytic activity, cell mediated immunity and humoralimmunity.Zinc enhances the phagocytic activity of macrophages andneutrophils. Phagocytic cell consume large quantities ofoxygen during the so-called respiratory burst, whichaccompanies the ingestion and killing of microorganisms andproduction of H

2O

2 and O-

2 radicals in response to challenge

by foreign particles. The protection of neutrophils againstthe damaging effects of super oxide radicals is probably thefunction of the cytosolic Cu-Zn containing SOD.

CopperCopper is a component of Superoxide dismutase enzyme.Through this enzyme activity, copper enhances thephagocytic process of neutrophils and macrophages.Ceruloplasmin a copper containing protein and around 90%the circulatory copper present in this form. The concentrationof ceruloplasmin higher in inflammatory site due to increasedblood supply. The copper present in the ceruloplasmin usedby neutrophils and macrophages used for their phagocytosisprocess.Copper deficient animals exhibit severe symptoms of immunedysfunctions like decreased functions of T cells, decreasedNK cell cytotoxicity and distorted lymphocyte population.Copper deficient animal show decrease in antibody cellresponse with increased susceptibility to infection. Copperdeficiency appears to alter the plasma membrane thus alteringimmune response to infection.


ChromiumChromium so important to health maintenance particularlyduring stress. Cr seems to be an essential trace elementbecause it is a component of Glucose Tolerance Factor (GTF)that potentiates the action of insulin. GTF is organo metalliccompound consist of trivalent chromium ions bound toseveral molecules of niacin, and amino acids. GTF facilitateinteraction between insulin and insulin receptor in targettissue.

Supplemental chromium enhances the immune responseof stressed calves. Stress result in elevated bloodconcentration of cortisol, which is known to depress immunefunction. Periparturient and early lactation dairy cows are undergreat physical and metabolic stress. Under these conditionsCr supplementation enhances immune responses. Chromiumsupplementation enhances both Humoral and cell mediatedimmune response under stress.Iron and CobaltIron is important for heam synthesis and it exerts immune rolevia catalase enzyme which converts hydrogen peroxide towater in anti oxidant system. Impaired cell mediated immuneresponse was observed in iron deficient animals. Primarilyaffect antibody formation associated with B cells. In pigs,iron deficiency prone to more disease susceptibility. Cobalt deficiency affects neutrophil function. Itsdeficiency affects resistance to parasitic functions. Higherfaecal egg counts are observed in Co deficient lambs afternatural infection with gastrointestinal nematodes.Vitamins as antioxidantsAntioxidant function as to remove harmful free radicalsproduced through normal cellular activity, there bymaintaining structural integrity of immune cells. Major freeradicals found in biological system are super oxide, hydrogenperoxide, hydroxyl radical and fatty acid radicals. Free radicalsare highly reactive compounds because they are missing anelectron. Free radicals can react with nucleic acids causingmutation, they can react with enzymes and render them inactive and they can react with fatty acids in membranescausing membrane instability. Free radicals can eventuallykill cells and damage tissue.

Reactive oxygen metabolites are unavoidable productsof normal metabolism process and are not ways harmful. Superoxide and hydrogen peroxide are involved physiologically inthe chemistry of several enzymes and are used by phagocyticcells to kill bacteria. In balance between production to reactiveoxygen metabolites and their safe disposal however caninitiate oxidative chain reactions and lipid per oxidation. Naturalantioxidant includes vitamin E, vitamin A carotenoids (betacarotene) and vitamin C. Beta carotene is potent direct actinganti oxidants where as vitamin A is less active anti oxidant butits role in disease resistance was well documented asmaintaining the epithelial integrity of immune cells.

Vitamin E and Selenium

Primary function of vitamin E is as an anti oxidant andits supplementation enhances the neutrophil function. Bothvitamin E and Se are important in cellular antioxidant system.High dietary vitamin E reduces the requirement for selenium.The principal biochemical role of selenium is through enzymeie. Glutathione peroxidase. Glutathione perxoidase is animportant part of cellular antioxidant system. Seleniumsupplementation also improves neutrophil function.Neutrophils from cows supplemented with 0.3 ppm ofsupplemental selenium killed mastitis pathogen moreeffectively than non-supplemented group.Vitamin E and Se influences the function of immune cellsespecially mammary gland. Vitamin E and the seleniumcontaining enzyme glutathione peroxidase are important inthe function on polymorphonuclear cells (PMN). Whenpathogen invades the mammary gland they trigger an influxof PMN and other white cells. These cells engulf and destroybacteria and other harmful organism. If vit E and Se are not inadequate supply, the total no. of PMN and the life span ofthese cells will be greatly reduced.Vitamin A and beta-caroteneCarotenoids are red and yellow pigments serve as precursor(beta carotene) to vitamin A. Beta-carotene is an efficientquencher of singlet oxygen and can function as antioxidant.Vitamin A cannot quench singlet oxygen and has lessantioxidant activity then other antioxidant. But vitamin Aprevents epithelial keratinization and maintains the cellularintegrity of lymphoid organs, which is important for combatingdisease stress.Beta carotene increases lymphocyte cytotoxic activity,stimulate production of various cytokines, enhancesphagocytic activity of neutrophils and macrophages andincrease activity of natural killer cells. It enhances both cellularand humoral immunity. Beta-carotene enhances peroxidaseactivity in macrophages and myelo peroxidase activity ofneutrophils.

Conclusion Micronutrients effectively modulate the animal immune

response. Zinc and copper enhance both cell mediated andhumoral immune response. Chromium evokes animal immunestatus especially in stress condition. Vitamins i.e. antioxidantsprevent the tissue against the free radicals generated in normalcellular metabolism. Vitamin E and Se particularly importantfor mammary gland immunity. Beta-carotene act as potentantioxidant and prevent the tissue damage caused by freeradicals.Thus, the possibility if dietary nutrient manipulation foroptimization of immune response with out compromising thegenetic potential of animals for growth and production appearsto be feasible and thus will economically benefit the livestockfarmers and the sector as whole.

U


STRESS – ITS ROLE IN REPRODUCTION IN PIGSU

INTRODUCTION

Commercial pig production presents the animals witha multitude of potentially stressful challenges. Distress isa threat to animal welfare and may impair productivity inboth growing and reproducing pigs. Deleterious effectstend to be neglected when intensifying production.Modern pig production is all about efficiency. Many factorsassociated with intensive rearing, such as crowding andmixing, are known to be stressful to animals. Althoughstress has the potential to decrease growth andreproduction in pigs, actions taken to reduce stress areoften neglected as an important aspect of economy.

Reproductive efficiency is the major affectingprofitability in many livestock production systems.Reproductive efficiency has a greater influence on theeconomic sustainability of a commercial livestockproduction than does any other performance traits. Thisis because reproductive efficiency is a composite trait thataffects the litter weight weaned and future meat production.Reproductive efficiency is an integrated measure of ageat puberty, capacity to produce and deliver adequatenumber of fertile spermatozoa, ovulation rate, ovumfertilization rate, embryo and foetal survival and ability tocope with a variety of environmental stressors.

Reduced reproductive efficiency can occur as a resultof environmental and management factors or stressorsassociated with animal housing, human-animal interaction,animal handling and management, modern productionmethods, temperature extremes. These stressors causedeviation in hormonal pattern and clinical manifestations.Reduction of stressful situations allow for greater well-being, growth and reproductive efficiency of the animal(NseAbasiet al., 2013).

STRESS

The stimuli that disrupt homeostasis are commonlytermed stressors and these can be physical, psychologicalor physiological (Dobson and Smith, 1995). Stress canbe defined as any environmental change; that is alteration

Supriya, S., Rajeshwari, Y. B., Sudharshan, V., Banuprakash, A.R. Anup Kumar, P. K.Department of Livestock Production and Management,Veterinary College Hebbal, Bangalore, KVAFSU, Bidar.

in climate or management that is severe enough to elicit abehavioural or physiological response from the animal(Coubrough, 1985).Moberg (1991), defined stress as abiological response elicited when an animal perceives athreat to its homeostasis.

Studies to determine the amount of stress on farmanimals are difficult to interpret from an animal welfarestandpoint. Animals can be stressed by either psychologicalstress; restraint, handling or novelty or physical stress:hunger, thirst, fatigue, injury or thermal extremes.Acutestress is a stress that lasts only brieûy (seconds, minutesor up to a few hours) and prolonged stress as stress whichis continuous (not repeated) and lasts much longer (days,weeks or months).

Problems of stress include induced changes in thesecretion of pituitary hormones, thus leading to alteredmetabolism, immune competence and behaviour, as wellas failure in reproduction. Under prolonged or extremestressful conditions, the effect of animal health can bevery significant resulting in irreversible losses inproductivity or even death. Chronic stress may affect thewelfare of the animal; affect the quality of the product.

NEUROENDOCRINE RESPONSE TO STRESS:

A number of physiological systems are activated inresponse to stress. Rapid responses to stressors aremediated by the sympathetic adrenal medullary systemwhich involves the central nervous system; these neuralpathways activate release of epinephrine by the adrenalmedulla and norepinephrine by peripheral sympatheticnerves. The hypothalamic-pituitary-adrenocortical (HPA)stress response system mediates a long term sustainedresponse with the involvement of major adrenal corticalhormones such as glucocorticoids and mineralocorticoids.Two classical stress response systems result in differenttemporal and context specific coping patterns wherebythe sympathetic nervous system is primarily activated insituations of threat, whereas HPA system is involved duringloss of control.


Perception of stressful stimuli leads to activation ofHPA system which in turn results in release of a variety ofpeptides, principally corticotrophin releasing hormone(CRH) and vasopressin from the hypothalamus. CRHstimulates the release of adrenocorticotrophic hormone(ACTH) and other propriomelanocortin derived peptidessuch as beta endorphin from anterior lobe of pituitarygland. ACTH acts on the adrenal gland and causes secretionof glucocorticoid hormones example cortisol. There isreduction in the amount of LH released by challenges withGnRH. The reduction in endogenous GnRH/LH secretionultimately deprives the ovarian follicle of adequategonadotrophin support leading to reduced oestradiolproduction by slower growing follicles.Elevations in theplasma concentrations of cortisol have commonly beenacknowledged as an indicator of when an animal isexperiencing stress. This has been utilized in many studieswith female pigs where plasma concentrations of cortisolhave been measured in response to various procedures.

Reproduction in females is controlled by the hormonalinteractions of the hypothalamo-pituitary-ovarian axiswhich involves the sequential release of gonadotropin-releasing hormone (GnRH) from the hypothalamus,luteinizing hormone (LH) and follicle stimulating hormone(FSH) from the anterior pituitary and the sex steroids,estrogen and progesterone from the ovaries. The seriesof precisely timed endocrine, behavioural and physiologicalevents of the estrous cycle are paramount for successfulreproduction in female pigs. It is essential that these eventsinduce ovulation with suitable timing relative to estrus sothat females can mate at a time appropriate for fertilization.Disruption of the series of endocrine events prior to estrusand ovulation is likely to inhibit reproduction and,consequently, it has been hypothesized that females maybe particularly susceptible to the effects of acute stressduring this pre-ovulatory period (Eberhardet al., 2007).

ASSESSMENT OF STRESS:

There are many difficulties involved in evaluation of,and comparing how different types of stress affect animalwelfare in general, especially in long-term stressfulsituation. The stress response is equally dependent on thenature, intensity and duration of the stressful event. Also,there is a large individual variation between pigs in theirability to cope with stress and the fact that each stressorboth has a non-specific effect and a specific effect(Einarssonet al., 2008).

Stress response can be assessed by determining theactivation of HPA-axis and/or the sympathetic adreno-medullary system, by measuring the levels of secretedpeptides in the peripheral blood plasma, urine, cerebrospinalfluid, saliva.Behavioural responses such as heart rate, bloodpressure and stereotypical behavior, as well as the effectson the immune response can also be used for theassessment of stress response. However meaningfulevaluation of these responses requires a detailed knowledgeof the normal physiological and behavior patterns of theanimals, because the response to stress is influenced byseveral factors such as the metabolic condition, healthstatus, age, sexual maturity.

STRESS AND REPRODUCTION:

Reproduction is the ultimate measure of an animal’sability to adapt to an ever-changing external environment,as well as forming the basis of life time productivity(Coubrough, 1985). Management induced stress isbecoming more important whenrelated to the requirementsof modern production methods.

According to Coubrough (1985), stressors causedeviations in hormonal pattern and clinical manifestations.Stressors affect reproductive functions through actionsat the hypothalamus as well as impairing pituitary LHrelease induced by GnRH (Dobson and Smith, 1995). Ifan animal is under stress during a critical period of theoestrus cycle (late proestrus or oestrus) a glucocorticoidinduced suppression of LH is likely to either delay orprevent ovulation and may reduce libido in males (Moberg,1976).

Physiological distress that can be caused bymovement of animals to new environments or caused byabusive treatment will elicit release of ACTH andglucocorticoids. Also, research has shown that embryosare more likely to be retarded and/or abnormal whencollected from female animals that were subjected to heatstress during estrus when compared to embryos fromthose that were not stressed. Another factor related to thelow fertility seen during heat stress is the evidence thatthe embryo loses its ability to alter prostaglandins synthesisin a manner that favours the maintenance of the corpusluteum when under such conditions. These effects,combined with the other endocrine changes which occurduring heat stress, accounts for the more pronouncedeffect of heat stress on reproduction than is seen withother stressors (Moberg, 2000).


In addition, there is evidence from both in vitro perifusionsand in vivo experiments to show that exogenously increasedACTH concentration or transport reduce the amount ofLH released by challenges with small doses of GnRH. Thisprovides support for additional effects at the pituitary level.Clearly, activation of the hypothalamus-pituitary-adrenalaxis by stressors reduces the pulsatility of GnRH-LHactions at both the hypothalamus and pituitary gland,ultimately depriving the ovarian follicle of adequate LHsupport. This will lead to reduced oestradiol productionby slower growing follicles. Such a hypothesis is supportedby the marked decrease in oestradiol secretion observedafter reducing the frequency of exogenous LH pulsesdriving follicular growth in an ovarian auto transplantmodel.

HEAT STRESS

Pigs are more susceptible to heat stress due to lessnumber of sweat glands, relatively smaller size of lungsand thick subcutaneous fat. Exposure of male and femalepigs to elevated ambient temperatures can result in reducedreproductive efficiency.

Effect onBoars :The effects of heat stress on semenquality appear about two weeks after heat stress isimposed, reach their maximum severity in 28-38 days andreturn to normal 5-8 weeks after heat stress ceases.Semenquality - lowered fertility / lowered total sperm counts inboars in summer.Negative effect on ejaculate volume, totalsperm count and morphology ofspermatozoa(Suriyasomboonet al.,2004).Increasedtemperature has an inhibitory effect on spermatidmaturation and on testicular androgenbiosynthesis(Wattemman, 1985).

Effect of heat stress on gilts/sow: Heat stress hasbeen reported to reduce implantation and impairembryo development in pig.Gilts are more sensitive to heatstress before day 15 of pregnancy.Reduction inthe number of viable embryos among gilts exposed toelevated temperatures during 8-16 days post-breeding –indicating time of implantation is more sensitive stage ofpregnancy to stress (Einarssonet al., 2008).

• Anoestrus : gilts reared during summer are commonlyobserved to be older and lighter at puberty.

• Wean to mate interval : Sows weaned during summeroften exhibit a delay in returning to oestrus.

• Duration and intensity of oestrus: Lowered by halfday in summer. Sexual interest and intensity alsolowered.

• Ovulation rate : Slightly reduced or no effect onovulation rate.

• Pregnancy : Embryo more vulnerable to heat stressduring implantation. Increased temperatures duringlast two weeks of pregnancy can cause increase instillbirths.

WEANING :

Comparative clinical and endocrine studies of sowsshowed, when all piglets were removed from their damswithin 12 hours of farrowing, called “zero weaning” andsows (showed estrus within 2 weeks after parturition)developed ovarian cysts (anovulatory). The peripheralplasma concentration of cortisol was significantly higherin the anovulatory sows than in ovulatory sows, indicatingthat elevated cortisol might be one of the factor inhibitingLH surge (Kunavongkritet al., 1984).

TRANSPORTATION :

Transport for 4 or 8 hours reduced the frequencyand amplitude of LH pulses especially within the first fewhours in ovariectomised ewes or intact animals in the latefollicular phase (Dobson et al., 1999). Plasmaconcentrations of both cortisol and catecholamines areelevated in pigs that are transported.Transport of femalepigs advanced the onset of puberty (Einarssonet al., 2008).Combined clinical and endocrinological studies have beenperformed on effect of transportation on gilts with delayedpuberty and anoestrus sows.75% of gilts and sows showedovulatory oestrus within one week, after one hour oftransportation.

Consequences of stress on reproductiveperformance :

Stress has an effect on puberty attainment, sexualbehavior, ovulation, embryo and foetal development,parturition and lactation.

Social and movement restriction due to tetheringinduces a chronic elevation in cortisol, but age at firstovulation are similar in tethered and group housed femalesreared in isolation from the boar.Acute stress associatedto transport, new environment, social mixing with orwithout boar exposure induces puberty in prepubertal


females.Stress induces plasma cortisol that continues forseveral hours but its role in stimulating puberty is not wellunderstood.

Overcrowding chronically increases plasma cortisol

and has negative influence on sexual behaviour.Shorter

duration of oestrus in tethered than in individual loose-

housed sows, but similar ovulation rate was

observed.Chronic elevation of corticosteroids after

repeated intramuscular injections of ACTH at an appropriate

period during oestrus cycle inhibits sexual behaviour.

Lower pregnancy rates occur in cyclic gilts submitted to

repeated electric shocks. Intense noise due to repeated

explosive detonations and construction work seems to

induce abortions in sows. Parturition and early lactation

are periods of profound behavioural and physiological

changes that are highly sensitive to stressors.Plasma

cortisol and ACTH increase in sows around parturition.

Plasma and salivary cortisol start to increase in around

12h before farrowing and remain elevated for about

24h.Increase in ACTH and cortisol is higher in sows housed

in crates without any bedding than in pens with straw.

Environmental disturbances such as moving the sow and

her litter to a new pen leads to unsuccessful nursings

probably due to lack of oxytocin release.

CONCLUSION :

Reproduction is a very important physiological

system for furtherance of a species and this has to succeed

despite the imposition of sometimes detrimental

environmental stimuli. To ensure that an animal can

respond to its surrounding, it is advantageous to have

several lines of defence, that is, higher brain, hypothalamus,

pituitary and adrenal glands. Likewise each of these

responses has influence on the different levels of the

reproductive organizations, that is, higher brain,

hypothalamus, pituitary and gonads.

Management practices to alleviate undesirable stress

involves recognizing and eliminating the cause of the stress

especially heat. Since reduced reproductive efficiency is

more dramatic and predictable during heat stress and may

include problems in detection of estrus, conception, and

fetal growth, a more basic understanding of the animal’s

response to heat is needed. This will help the animal

manager adopt practices to increase reproductive efficiency

during hot weather/climate.

References :

COUBROUGH, R.I. 1985. Stress and fertility.Onderspoot

J. Vet. Res. 52 : 153-156.

DOBSON, H., SMITH,R.F. 1995. Stress and reproduction

in farm animals.J. Reprod. Fertil.Suppl.49 : 451-461.

DOBSON, H., TEBBLE, J.E., PHOGAT, J.B., SMITH,

R.F.1999. Effect of transport on pulsatile and surge

secretion of LH in ewes in the breeding season.J. Reprod.

Fertil. 116: 1–8.

EBERHARD, V.B., DOBSON,H. and PRUNIER,A. 2007.

Stress, behavior and reproductive performance

in female cattle and pigs.Horm.Behav. 50 : 130-138.

EINARSSON,S., BRANDT,Y., LUNDEHEIM, N. and

MADEJ,A. 2008. Stress and its influence on

reproduction in pigs a review.Acta. Vet. Scand. 50 : 48.

KUNAVONGKRIT,A., MADEJ,A. and EINARSSON,S.

1984. Plasma levels of cortisol in zero-weaned and

lactating sows during the first two weeks post partum.

Domestic Animal Endocrinology.1 : 217-223.

MOBERG,G.P. 1991. How behavioral stress disrupts the

endocrine control of reproduction in domestic

animals. J. Dairy. Sci. 74 : 304-311.

NSEABASI ETIM,N., EDEM, E. A., METIABASI

UDO,D., MARY WILLIAMS,E., and EMEM EVANS,I.

2013. Physiological relationship between stress and

reproductive efficiency.Agric. Biol. J. N. Am. 4 (3) :

600-604.

SURIYASOMBOON,A., LUNDEHEIM,N.,

KUNAVONGKRIT,A., ans EINARSSON,S. 2004.

Effect of temperature and humidity on sperm production

in Duroc boars under different housing systems in

Thailand. Livest. Prod. Sci. 89 : 19-31.

WATTEMMAN,R.P., BAZER,F.W. 1985. Influence of

environmental temperature on prolificacy of pigs.

J.Reprod.Fertil. Suppl. 33 : 199-208.

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Transport Myopathy of Turkeys(Leg Edema Syndrome) - A skeletal Disorder

U

Introduction

Turkey leg oedema is a syndrome which may on occasion

affect a high percentage of birds from a given lot and have

severe economic consequences in terms of condemned

parts. Even though the syndrome is well-known, it has

not been subject to scientific study. Heavy toms are pri-

marily affected, although it also develops in hens. About

5% of all flocks are affected, and morbidity within the

flock is 2-20% but can occasionally be as high as 70%.

Transport myopathy occurs sporadically but is most com-

mon during fall and early winter. A high incidence has oc-

curred in sequential flocks from the same farm. Incidence

is likely to be higher in flocks raised in confinement than in

range flocks.

Etiology

The cause is unknown, but transport myopathy is associ-

ated with increased body size and weight, increased trans-

port time to processing plant, cool ambient temperatures,

and valgus leg deformities. The pathogenesis is unknown

but presumed to be similar to exertional myopathy.

Symptoms and Diagnosis

Often only one leg is affected. No evidence of external

trauma is seen. Skin over edematous subcutaneous tissue

is pale, feather follicles are less visible, and the skin slips

easily over underlying muscle when moved. Occasionally,

there is crepitation. Affected areas are dark when the edema-

tous areas contain blood. Typically, when the edematous

areas are cut, the subcutis is a few to several millimeters

thick and is amber, occasionally green, or rarely red. Both

the pectoralis thoracicus and biceps femoris are affected,

with the former to a slightly greater extent. Purulent exu-

date is absent, which distinguishes transport myopathy

from cellulitis. The lesions can be monophasic (resulting

from a single event, eg, transport, capture, or restraint

myopathy) or polyphasic (with repeated or ongoing events).

Fasciculi were often small and widely separated from each

other by large amounts of proliferated perimysial connec-

tive tissue. Great variation in the size of individual fibres

can be observed and nuclei are usually shrunken and py-

knotic. If hemorrhage is present, the adductor muscle is

usually torn. Removal of affected legs at processing re-

sults in carcass downgrading. Microscopically, acute mul-

tifocal muscle necrosis is found, primarily in the adductor

muscles. Sometimes subacute or chronic lesions are seen,

suggesting earlier episodes of myopathy. Serum CK in-

creases sharply between farm and processing.

Remedial Measures

The derangements of ante-mortem muscle cell metabo-

lism and alterations in sarcolemmal integrity and tissue

structure associated with the presence of myopathy may

have profound implications for meat quality and the inci-

dence of specific conditions such as Pale, Soft Exudative

(PSE)-like meat. Therefore, it is necessary that the pro-

grams be designed to improve leg strength and conforma-

tion and to reduce trauma during transportation that will

help to reduce the incidence of this myopathy. Supple-

mental vitamin E and Selenium too may be useful. If pos-

sible, flocks with a high incidence of valgus leg deformi-

ties should be marketed early at the processing plant nearby.

U

Aasif Ahmad Sheikh*1, Showkat A. Bhat1, Mohammad Rayees Dar1,Thulasiraman Parkunan1, Lakshmi Priyadarshini1 and Hilal Ahmad Rather2

1National Dairy Research Institute, Karnal, Haryana, INDIA2Indian Veterinary Research Institute, Mukhteshwar Campus, Uttrakhand, INDIA

*Corresponding Author; Email ID:[email protected]



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