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English Editor

Yanka Ivanova (Bulgaria)

2015

ISSN 1313 - 8820 Volume 7, Number 3September 2015

Histopathological changes in small intestines of broiler chickens with experimental aflatoxicosis

1Department of General and Clinical Pathology, Faculty of Veterinary Medicine, Trakia University, 6000 Stara Zagora, Bulgaria2Department of Internal Non-infectious Diseases, Faculty of Veterinary Medicine, Trakia University, 6000 Stara Zagora, Bulgaria

1 2 2 2 2N. Grozeva *, I. Valchev , Ts. Hristov , L. Lazarov , Y. Nikolov

Abstract. Aflatoxins are toxic metabolites produced by moulds of Aspergillus genus (the most relevant producers are Aspergillus flavus and Aspergillus parasiticus). The primary aim of the current research is to follow out the toxic effects of aflatoxin В on the morphology of small intestine, which is important for 1

digestion and absorption of feeds and at the same time, to investigate the possibility for prevention of toxic AFB effects by addition of the mycosorbent Mycotоx 1

NG (Ceva Sante Animale, France) to the feed. The experiments were carried out with 50 Cobb broiler chickens, divided in 5 groups of 10 chickens (5 male : 5 female in the groups): Group I – control (receiving feed according to species and age); Group II – supplemented with 1 g/kg Mycotox NG; Group III – supplemented with 0.5 mg/kg aflatoxin B ; Group IV – supplemented with 0.8 mg/kg aflatoxin B ; Group V – supplemented with 0.5 mg/kg aflatoxin B and 1 g/kg 1 1 1

Mycotox NG. The experiment's duration was 42 days. After conventional histological technics the light microscopical analyze determined that the morphological changes in all small intestine (duodenum, jejunum, ileum) in birds treated at 0.5 mg/kg AFB consisted in vascular hyperaemia, single 1

haemorrhagies, partial desquamation of villous epithelium, rarefaction of cells in intestinal lymphatic follicles and in some areas, cup cell hypertrophy and weak distrophy of Lieberkühn glands. Chickens treated with 0.8 mg/kg AFB showed more intense dystrophic changes - necrobiotic to necrotic changes of intestinal 1

villi, atrophy of Lieberkühn glands, mononuclear cell infiltration of the propria and severe rarefaction of lymphatic follicular cells. The dietary supplementation of chickens with Mycotox NG resulted in fewer and less severe histological changes – no haemorrhagies but only vascular hyperaemia.

Keywords: aflatoxin В , small intestine lesions, broiler chickens, Mycotox NG1

AGRICULTURAL SCIENCE AND TECHNOLOGY, VOL. 7, No 3, pp , 2015313 - 318

Introduction disease and stress and lower weight gain (Coelho, 1990). Aflatoxin В is the most toxic compound (Busby and Wogan, 1981; Кubena et 1

al., 1990; Hsieh and Atkinson, 1991; Barjesteh et al., 2010), with The contamination of poultry feeds with mycotoxins is a serious strong hepatotoxic, hepatocarcinogenic, teratogenic, cytotoxic and concern for global poultry industry. The profitability of poultry farming mutagenic effects (Mishra and Das, 2003; Mehrim et al., 2006). The could be substantially reduced by the frequent contamination of intake of AF-contaminated feeds results in lower live weight, lower feeds with mycotoxins from one hand, and consequently to the weight gain, feed intake, increased feed conversion (Abousadi et al., harmful effects of mycotoxins on production traits on the other 2007; Zao et al., 2010; Yildirim et al., 2011), altered relative weights (Hamilton, 1984). of visceral organs (Teleb et al., 2004; Manafi et al., 2012), altered Aflatoxins are the most popular and most intensively studied morphology of the liver (Zao et al., 2010; Yildirim et al., 2011), mycotoxins in the world. Among the eighteen indentified aflatoxins kidneys (Mohamed and Mohamed, 2009; Yildirim et al., 2011), (AF), AFB , AFB , AFG , and AFG are the principal four forms found 1 2 1 2

immune organs – thymus, bursa of Fabricius, spleen (Sakhare et al., in naturally contaminated feeds (Tedesco et al., 2004). АF are a 2007) and intestines (Jakhar and Sadana, 2004; Mehrim et al., 2006; groups of closely related bioactive mycotoxins produced by fungi Aboutalebi, 2013). from the Aspergillus genus (A. flavus and A. parasiticus) (Wilson and

A EC report states that 30% of feeds and agricultural products Payne, 1994; Celyk et al., 2003). They are commonly found as are contaminated with mycotoxins, 90% of them being aflatoxins. To poultry feed contaminants (Edds and Bortell, 1983). A. flavus is one decrease or remove mycotoxins from the feeds, a variety of of the commonest contaminants of cereal crops used in animal and chemical, physical and biological methods have been proposed human nutrition (Abarca et al., 1994). AF are mainly produced during (Santin et al., 2003; El-Nezami et al., 2004; Abbès et al., 2006a,b). the storage of cereals or feeds and less frequently encountered The removal of AF from contaminated feeds is a main issue in the before the harvest. These mycotoxins are relatively heat-stable and quest of efficient decontamination technologies (Oguz et al., 2000). are frequently detected after technological processing and Most methods are expensive, inefficient and potentially dangerous. packaging. In animal feeds, AF production is enhanced at ambient At present, the most popular method for feed decontamination is the temperatures above 27°С, and feed humidity above 14% (Royes use of inorganic mycosorbents (natural and synthetic bentonites and and Yanong, 2002). Growing turkeys, quails, ducklings, goslings and zeolites). They bind aflatoxins during the digestion and thus, reduce chickens are particularly sensitive to the toxic effects of AF (Arafa et their absorption from the gastrointestinal tract (Ramos et al., 1996; al., 1981). Fatal outcome is observed only after ingestion of relatively Kececi et al., 1998; Oguz and Kurtoglu, 2000; Ibrahim et al., 2000; high amounts. At the same time, low amount of AF ingested with Miazzo et al., 2000; Rosa et al., 2001).feeds could be deleterious if the intake is over a prolonged period of

The aim of the current research is to follow out the toxic effects time. For growing birds, feed should contain no more than 20 μg/kg of aflatoxin В on the morphology of small intestine, which is 1aflatoxins. Yet, AF levels <20 μg/kg result in reduced resistance to

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* e-mail: dr_grozeva@yahoo.com

important for digestion and absorption of feeds. At the same time, in non-supplemented chickens as well as in chickens treated with 1 the possibility for prevention of toxic AFB effects by addition of the g/kg Mycotox NG in the feed. 1

Chickens supplemented with 0.5 mg/kg AFB and 1 g/kg feed mycosorbent Mycotоx NG (Ceva Sante Animale, France) to the feed 1

was investigated. Mycotox NG, exhibited a mild swelling and reddening of mucosa in some small intestine areas, the duodenum being the most affected. In the jejunum and ileum, the changes gradually subsided.

The mucosa of all small intestine compartments of chickens Material and methodstreated with 0.5 or 0.8 mg/kg AFB was swollen and reddened, 1

spattered with petechial or striated haemorrhagies in some locations The experiments were carried out with 50 Cobb 7-day-old (Figure 1). In the region of the duodenum and the jejunum, it was broiler chickens, divided in 5 groups of 10 chickens (5 male : 5 covered by a profuse amount of mucous descvamatives and female). secretes. · Group I – control, the chickens received compound feed

according to their age, produced at Provimi, Stara Zagora;istopathological studies· Group II – experimental, supplemented with 1 g/kg Mycotox

Chickens treated with 0.5 mg/kg AFB exhibited generalised 1 NG; hyperaemia in all small intestine compartments (duodenum, · Group III – experimental, supplemented with 0.5 mg/kg jejunum, ileum), at some areas with single haemorrhages, the aflatoxin B ; 1duodenum being the most severely affected. A part of villous · Group IV – experimental, supplemented with 0.8 mg/kg epithelium was desquamated and shed into the lumen (denuding of

aflatoxin B ; 1 villi). In some intestinal lymphatic follicles, rarefaction of cells and · Group V – experimental, supplemented with 0.5 mg/kg cup cell functional hypertrophy was seen. Weak dystrophy was

aflatoxin B and 1 g/kg Mycotox NG. 1 observed also in the Lieberkühn glands – disintegration and Aflatoxin В produced by Aspergillus flavus (99% purity) was 1 desquamation of basal membrane cells, with lysis and pyknosis of

purchased from Sigma-Aldrich, Germany. All birds were housed nuclei (Figure 2). under identical optimum microclimatic parameters as required by In chickens receiving 0.8 mg/kg AFB with their feed, the small 1

Ordinance 44/2006. intestine dystrophy was more intense. There were necrobiotic to All stages of the experiment were appryoved by the Trakia necrotic changes in intestinal villi (apart denuded villi, entire areas

University (Stara Zagora) Animal Etics Committee (Permit № where necrotic villi have shed into the lumen could be observed). At 49/29.09.2012) the same time, Lieberkühn glands also have underwent atrophic

After the end of the experiment, the birds were euthanised by changes. In the small intestine propria, mononuclear cell infiltration cervical dislocation. Small intestine specimens were fixed in 10% and severe rarefaction of lymphatic follicular cells was detected formalin and routinely processed. Specimens were embedded in (Figures 3 and 4). paraffin blocks, sectioned at 3 µm on a Leica RM 2235 microtome, In chickens treated with 0.5 mg/kg AFB and 1 g Mycotox NG, 1 and stained with haematoxylin/eosin. small intestine dystrophic changes were substantially less

pronounced than in groups supplemented with 0.5 and 0.8 ppm AFB , with mononuclear cell infiltration of the propria and weaker 1

Results dystrophic changes in intestinal lymph follicles as main findings. The villous and glandular dystrophy intensity was reduced, no haemorrhagies were present, but only vascular hyperaemia (Figure

There were no macroscopic changes in small intestinal mucosa 5).

Gross anatomy studies

ure Hyperaemia, petechial and striated haemorrhagies in the intestines of broiler chickens after 42-day dietary supplementation with 0.8 mg kg AFВ1

ure 2 es uamation of illous epithelium and sheddinginto the intestinal lumen in chickens after 42-day dietary supplementation with 0.5 mg kg AFВ . H . ar 15 µm1

digestion and absorption of nutrients. The dystrophic changes in Lieberkühn glands within the small intestine are the cause for reduced secretion of various enzymes (saccharose and maltose) involved in digestion (Aboutalebi, 2013). The gastrointestinal tract is the main route through which the aflatoxins enter the animal body after consumption of AFB –contaminated feeds. At the same time, 1

the digestive tract is the main route of excretion of AFB metabolites 1

through the bile. Aflatoxins and their main metabolites AF-8,9-epoxides cause carcinomas of small and large intestines in men (Agag, 2004) and animals (Coulombe, 1994). It is demonstrated that aflatoxins cause also various acute clinical and morphological effects on the gastrointestinal tract, such as vomiting, diarrhoea, gastrointestinal bleeding etc. (Harriet, 2003).

Small intestinal morphological changes in chickens from experimental group III (treated with 0.5 mg/kg AFB ) provided proofs 1

for catarrhal enteritis and Lieberkühn glands dystrophy. Chickens from group IV supplemented with higher amounts of AFB (0.8 mg/kg 1

AFB ) showed more severe histopathological changes (necrobiotic 1

to necrotic alterations of villi, mononuclear cell proliferation, rarefaction of lymphatic follicles). These changes are similar to inflammatory and dystrophic changes reported by other researchers

Neither control chickens nor experimental birds receiving only 1 in broiler chickens (Kumar and Balachandran, 2009), quails (Jakhar g/kg Mycotox NG with the feed showed gross morphology and Sadana, 2004), growing hens (Jayabarathi and Mohamudha alterations of small intestinal mucosa. Parveen, 2010) and fish (Abdelhamid et al., 2002; 2004). In broiler

chickens, catarrhal enteritis with lymphocytic and mononuclear cell proliferation was reported after feeding rations contaminated with 1 ppm AF for 4 weeks (Kumar and Balachandran, 2009). In quails, DiscussionJakhar and Sadana (2004) observed catarrhal enteritis demonstrated by increased cup cell number, congestive events in Histopathological changes in small intestine mucosa are the mucosa, haemorrhagies, necroses, lymphoid cell infiltration. In directly related to the amount of ingested AFB , the duration of 1

growing hens supplemented with 100 ppb total aflatoxin, vacuolation exposure (treatment) and the animal or avian species (Jakhar and of mesenchymal cells of intestinal propria has been observed Sadana, 2004). It is acknowledged that mycotoxins are absorbed (Jayabarathi and Mohamudha Parveen, 2010). Fish fed 500 – 2000 through the small intestine at a very high rate. According to reported ppb AFB with the feed showed oedema, mononuclear cell 1data, a complete absorption of aflatoxins in small intestine could infiltration and increased cup cell numbers (Abdelhamid et al., occur (Ramos et al., 1996). Aflatoxin В is the most toxic among all 1

2002). In another study in fish which received 100 – 200 ppb AFB1 indentified aflatoxins. It is established that aflatoxins disturb the Abdelhamid et al., (2004) reported impairment of the normal normal morphology of intestines (Jaeschke et al., 2002). intestinal architectonics and rarefaction of intestinal villi. Pathomorphological investigations showed that aflatoxin В exerted 1

The intestinal mucous haemorrhagies observed by us and by a most potent toxic effect on the duodenum, which is essential for

ure 3 ononuclear cell infiltration in the small intestine

propria (→) and hypersecretion from cup cells (∙∙∙ ) in

chickens after 42-day dietary treatment with 0.8 mg kg AFВ . H . ar 15 µm1

→ure 4 e ere rarefaction of small intestine lymphatic

follicular cells in chickens after 42-day dietary supplementation with 0.5 mg kg AFВ . H . ar 15 µm1

ure 5 ascular hyperaemia and weak dystrophic changes of intestinal illi and glands in chickens after 42-day treatment with 0.5 mg kg AFВ and 1 g ycoto 1

. H . ar 20 µm

316

others (Rumbeiha, 2001; Harriet, 2003) are due to biochemical Bacha H and Othman O, 2006a. "The protective effect of hydrated disturbances of the coagulation cascade induced by aflatoxins. They sodium calcium aluminosilicate against haematological, are characterised by reduction of thromboplastin, plasma biochemical and pathological changes induced by zearalenone in prothrombin, fibrinogen and factors V, VII and X (Tung et al., 1972; mice". Toxicon, 47, 567-574.Witlock and Wyatt, 1978). Doerr et al. (1976) proved the interference Abdelhamid AM, Abdel- Khalek AE, Mehrim AI and Khalil FFM, of aflatoxins with the intrinsic coagulation system observing decline 2004. An attempt to alleviate aflatoxicosis on Nile tilapia fish by in factors VIII and IX levels. Lower levels of all blood clotting factors dietary supplementations with chickenhatchery by-products (egg induced the haemorrhages in the different intestinal compartments. shells) and shrimp processing wastes (shrimp shells). 2- On clinical, Doerr et al. (1974) outlined extrinsic coagulation pathway disorders blood and histological parameters. Journal of Agriculture Scientific which prolonged prothrombin and blood clotting times as a cause for Mansoura University, 29, 6175-6196.occurring coagulopathies in different internal organs following Abdelhamid AM, Khalil FFM, El-Barbary MI, Zaki VH and feeding AF-contaminated feeds to broiler chickens. Aflatoxins inhibit Hussein HS, 2002. Feeding Nile tilpaia on Biogen® to detoxify protein synthesis, form adducts with DNA, RNA and proteins, inhibit aflatoxic diets. Proc.1st Conf. Animal and Fish Prod., Mansoura, 24-RNA synthesis through binding to DNA-dependent RNA, 25, Sept.,207-230.degranulate the endoplasmatic reticulum and thus, provoke Abousadi AM, Rowghani E and Ebrahimi Honarmand M, 2007. structural changes in a number of tissues (liver, kidneys, skeletal The efficacy of various additives to reduce the toxicity of aflatoxin B 1

muscles, intestines, heart) (Clifford and Rees, 1967; Wangikar et al., in broiler chicks. Iranian Journal of Veterinary Research, 8, 144-150.2005; Mohammed and Metwally, 2009; Sharma et al., 2011). Aboutalebi N, 2013. Toxic Effects of Aflatoxin B on Duodenum 1

Reduced severity of intestinal pathomorphological changes in Tissue. Journal of American Science, 9, 115-117.fish have been observed after addition of some nutritional Agag BI, 2004. Mycotoxins in foods and feeds : Aflatoxins. supplements such as ginger to AF-contaminated feeds (Mehrim et Association of Universal Bullettin of Environmental Research, 7, al., 1996; Platel and Srinivasan, 1996; 2000; Ahmed and Sharma, 173-191.1997). Comparable results are demonstrated also in the present Ahmed RS and Sharma SB, 1997. Biochemical studies on study. The addition of 1 g/kg Mycotox Ng to the feed of experimental combined effects of garlic (Allium sativum Linn) and ginger (Zingiber group V partly decreased the severe toxic effect of AB on small 1 officinal Rose) in albino rats. Indian Journal of Experimental Biology, intestine morphology (lower intensity of villous. glandular and 35, 841.lymphatic follicles' dystrophy). Arafa AS, Bloomer RJ, Wilson HR, Simpson CF and Harms RH,

1981. Susceptibility of various poultry species to dietary aflatoxin. British Poultry Science, 22, 431-436.Barjesteh MH, Azizi LG and Noshfar E, 2010. Occurrence of Conclusionaflatoxin M in pasteurized and local yogurt in Mazandaran province 1

(Northern Iran) using ELISA. Global Veterinaria, 4, 459-462.In conclusion, the independent inclusion of increasing levels of Busby WF and Wogan GN, 1981. Aflatoxins. Pages 3–27 in: AFB – 0.5 or 0.8 mg/kg (experimental groups III and IV) in 1Mycotoxins and N-Nitrosocompounds, Environmental Risks. Vol. 2. compound feed of broiler chickens resulted in altered small intestine R. C. Shank, ed. CRC Press Inc., Boca Raton, FL.morphology. The severity of histopathological changes in our Çelýk KL, Muzaffer D and Türker S, 2003. Reduction of Toxic experiment correlated to the tested AFВ amount. In experimental 1 Effects of Aflatoxin B1 by Using Baker Yeast (Saccharomyces

group III, the observed histopathological changes were less severe cerevisiae) in Growing Broiler Chicks Diets. Revista Brasileira de

(hyperaemia, haemorrhagies, desquamation of villous epithelium, Zootecnia, 32, 615-619.

rarefaction of cells in lymphatic follicles, cup cell hypertrophy, weak Clifford JI and Rees KR, 1967. The Interaction of Afiatoxins with

dystrophy in Lieberkühn glands). In experimental group IV, he Purines and Purine Nucleosides. Biochemistry Journal, 103, 467-

changes were more pronounced (necrobiotic to necrotic alterations 471.

of villi, atrophy of Lieberkühn glands, proliferation of mononuclear Coelho MB, 1990. Molds, mycotoxins and feed preservatives in the

infiltrates, rarefaction of lymphatic follicles). The addition of 1 g/kg feed industry. BSAF Corparation, 100. Cherry Hill Road, Parsippany,

Mycotox NG to the ration containing 0.5 mg/kg AFВ succeeded to 1 NJ07054, 1990. p.159.reduce partly the pathomorphological lesions due to aflatoxicosis

Coulombe RA, 1994. Nonhepatic disposition and effects of (lower intensity of villous and glandular dystrophy, lack of

aflatoxin B . The Toxicology of Aflatoxins: Human Health, Veterinary 1haemorrhages, presence of vascular hyperaemia only).and Agricultural significance.,http://toxicology.usu.edu/endnote/Nonhepatic-disposition.pdf., 89-101.Doerr JA, Huff WE, Tung HT, Wyatt RD and Hamilton PB, 1974. ReferencesSurvey of T-2 toxin, ochratoxin and aflatoxin for their effects On the coagulation of blood in young broiler chickens. Poultry Science, 53, Abarca ML, Bragulat MR, Castella G and Cabanes FJ, 1994. 1729-1734.Mycoflora and aflatoxin-producing strains in animal mixed feeds. Doerr JA, Wyatt RD and Hamilton PB, 1976. Impairment of Journal of Food Protection, 57, 256-258.coagulation function during aflatoxicosis in young chickens. Abbès S, Ben Salah-Abbès J, Ouanes Z, Houas Z, Othman O, Toxicology and Applied Pharmacology, 35, 437-446.Bacha H, Abdel-Wahhab MA and Oueslati R, 2006b. "Preventive Edds GT and Bortell RR, 1983. Biological effects of aflatoxin in role of phyllosilicate clay on the immunological and biochemical poultry. Pages 55–61 in: Aflatoxin and Aspergillus flavus in Corn. U. toxicity of zearalenone in balb/c mice". International Journal of L. Diener, R. L. Asquith, and J. W. Dickens, ed. Southern Immunopharmacology, 6, 1251-1258.Cooperative Services Bulletin. 279. Alabama Agricultural Abbès S, Ouanas Z, Ben Salah-Abbès J, Houas Z, Oueslati R,

317

Experiment Station, Auburn University, AL.. 16.El-Nezami H, Polychronaki N, Lee Y, Haskard C, Juvonen R, Oguz H and Kurtoglu V, 2000. Effect of clinoptilolite on Salminen S and Mykkanen H, 2004. "Chemical moieties and performance of broiler chickens during experimental aflatoxicosis. interactions involved in the binding of zearalenone to the surface of British Poultry Science, 41, 512-517. lactobacillus rhamnosus strains GG" Journal of Agricultural and Oguz H, Kececi T, Birdane YO, F. Onder F and Kurtoglu V, 2000. Food Chemistry, 52, 4577-4581. Effect of clinoptilolite on serum biochemical and haematological Hamilton PB, 1984. Determining safe levels of mycotoxins. Journal characters of broiler chickens during experimental aflatoxicosis. of Food Protection, 45, 570-575. Research in Veterinary Science, 69, 89-93.Harriet AM, 2003. Is indoor mold contamination a threat to health? Ordinance 44/20.04.2006 for veterinary medical requirements to Journal of Environmental Health, http://130.88.242.202/medicine/ animal rearing facilities, Official Gazette 41, Appendix 7.Aspergillus/articlesoverflow/ 12971049. pdf, 62, 2, 0022-0892. Platel K and Srinivasan K, 1996. Influence of dietary spices or their Hsieh DPH and Atkinson DN, 1991. Bisfuranoid mycotoxins:Their active principles on pancreatic digestive enzymes of small intestinal genotoxicity and carcinogenicity. Advances in Experimental mucosa in rats. International Journal of Food Science and Nutrition, Medicine and Biology, 283, 525-532. 47, 55.Ibrahim IK, Shareef AM and Al-Joubory KMT, 2000. Ameliorative Platel K and Srinivasan K, 2000. Influence of dietary spices and effects of sodium bentonite on phagocytosis and Newcastle disease their active principles on pancreatic digestive enzymes in albino rats. antibody formation in broiler chickens during aflatoxicosis. Research Nahrung, 44, 42.in Veterinary Science, 69, 119-2. Ramos AJ, Fink-Gremmels J and Hernandez E, 1996. Prevention Jaeschke H, Gores GJ, Cederbaum AI, Hinson JA, Pessayr D of toxic effects of mycotoxins by means of onnutritive adsorbent and Lemasters JJ, 2002. Mechanisms of hepatotoxicity. compounds. Journal of Food Protection, 59, 631-641.Toxicological Science, 65, 166-176. Rosa CAR. Miazzo R, Magnoli C, Salvano M, Chiacchiera SM, Jakhar KK and. Sadana JjR, 2004. Sequential pathology of Ferrero S, Saenz M, Carvalho ECQ and Dalcero A, 2001. experimental aflatoxicosis in quail and the effect of selenium Evaluation of the efficacy of bentonite from the South of Argentina to supplementation in modifying the disease process. Mycopathologia, ameliorate the toxic effects of aflatoxin in broilers. Poultry Science, 157, 99-109. 80, 139-144.Jayabarathi P and Mohamudha Parveen R, 2010. Biochemical Royes JB and Yanong RP, 2002. "Molds in fish feeds and and histopathological analysis of aflatoxicosis in Growing hens fed aflatoxicosis." Copyright by the University of Florida, Institute of with commercial poultry feed. International Journal of Agricultural Science (UF/ IFAS).Pharmaceutical Sciences Review and Research, 3, 127-130. Rumbeiha W, 2001. Mycotoxicosis in pets. DCPAH-Newsletter,18, Kececi T, Oguz H, Kurtoglu V and Demet O, 1998. Effects of 2.polyvinylpolypyrrolidone, synthetic zeolite and bentonite on serum biochemical and hematological characters of broiler chickens during aflatoxicosis. British Poultri Science, 39, 452-458.Kubena LF, Harvey RB, Huff WE, Corrier DE, Phillips TD and Rottinghaus GE, 1990. Efficacy of a hydrated sodium calcium aluminosilicate to reduce the toxicity of aflatoxin and T-2 toxin. Santin E, Paulilo AC, Maiorka A, Nakaghi LSO, Macan M, de Poultry Science, 69, 1078-1086. Silva AVE and Alessi AC, 2003. Evaluation of the efficiency of

Saccharomyces cerevisiae cell wall to ameliorate the toxic effects of aflatoxin in broilers. International Journal of Poultry Science, 2, 241-344.

Manafi M, Umakantha B, Noor Ali M and Narayana Swamy HD, Sharma V, Sharma C, Paliwal R, Pracheta and Sharma S, 2011. 2012. Study of the Combination Effects of Aflatoxin and T-2 Toxin on Ameliorative Effects of Curcuma Longa and Curcumin on Aflatoxin Performance Parameters and Internal Organs of Commercial B Induced Serological and Biochemical Changes In Kidney of Male 1

Broilers. Global Veterinaria, 8, 393-396. Mice. Asian Journal of Biochemical and Pharmaceutical Research, Mehrim AI, Abdelhamid AM, Abo Shosha AAM, Salem MF and 1, 2, 338-351El-Sharawy MAMM, 2006. Nutritious Attempts to Detoxify Aflatoxic Tedesco D, Steidler S, Galletti S, Tameni M, Sonzogni O and Diets of Tilapia Fish 2- Clinical, Biochemical and Histological Ravarotto L, 2004. Efficacy of silymarin-phospholipid complex in Parameters. Journal of the Arabian Aquaculture Society, 1, 69-90. reducing the toxicity of aflatoxin B in broiler chicks. Poultry Science, 1

Miazzo R, Rosa CA, De Queiroz Carvalho EC, Magnoli C, 83, 1839-1843.Chiacchiera SM, Palacio G, Saenz M, Kikot A, Basaldella E and Teleb HM, Hegazy AA and Hussein YA, 2004. Efficiency of Kaolin Dalcero A, 2000. Efficacy Of Synthetic Zeolite To Reduce The and Activated Charcoal to Reduce the Toxicity of Low Level of Toxicity Of Aflatoxin In Broiler Chicks. Poultry Science, 79, 1-6. Aflatoxin in Broilers. Scientific Journal of King Faisal University,

(Basic and Applied Sciences), 5, 145-159.Tung HT, Donaldson WE and Hamilton PB, 1972. Altered lipid transport during aflatoxicosis. Toxicology and Applied

Mohamed MA and Mohamed MH, 2009. Haemato-biochemical Pharmacology, 22, 97-104.and pathological studies on aflatoxicosis and treatment of broiler Wangikar PB, Dwivedi P, Sinha N, Sharma AK and Telang AG, chicks in Egypt. Veterinaria Italiana, 45, 323-37. 2005. Teratogenic effects in rabbits of simultaneous exposure to Mohammed AM and Metwally NS, 2009. Antiaflatoxicogenic ochratoxin A and aflatoxin B with special reference to microscopic 1

activities of some aqeous plant extracts against AFB induced Renal 1 effects. Toxicology, 215, 37-47.and Cardiac damage. Journal of Pharmacology and Toxicology, 4, 1- Wilson DM and Payne GA, 1994. Factors affecting Aspergillus

Kumar R and Balachandran C, 2009. Histopathological changes in broiler chickens fed afl atoxin and cyclopiazonic acid. Veterinarski Arhiv, 79, 31-40.

Mishra HN and Das C, 2003. A review on biological control and metabolism of aflatoxin. Critical Reviews in Food Science and Nutrition, 43, 245-264.

Sakhar PS, Harne SD, Kalorey DR, Warke SR, Bhandarkar AG and Kurkure NV, 2007. Effect of Toxiroak® polyherbal feed supplement during induced aflatoxicosis, ochratoxicosis and combined mycotoxicoses in broilers, Veterinarski Arhiv, 77, 129-146.

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Flavus group infection and aflatoxin contamination of crops. In: Eaton, D.L. Groopman, J.D. (Eds.). The toxicology of Aflatoxins. Academic Press inc, San Diego, pp123-145.Witlock DR and Wyatt RD, 1978. Effect of Eimeria tenella infection Zhao J, Shirley RB, Dibner JD, Uraizee F, Officer M, Kitchell M, and dietary aflatoxin on blood coagulation of young broiler chicks. Vazquez-Anon M and Knight CD, 2010. Comparison of hydrated Avian Diseases, 22, 481-486. sodium calcium aluminosilicate and yeast cell wall on counteracting

aflatoxicosis in broiler chicks. Poultry Science, 89, 2147-2156.

Effects of yeast lucomannan on performance, some biochemical parameters and pathological changes in experimental aflatoxicosis in broiler chickens. Revue de Médicine Véterinaure, 162, 413-420.

Yildirim E,Yalchinkaya I, Kanbur M, Çnar M and Oruc E, 2011.

Review

Genetics and Breeding

Nutrition and Physiology

Production Systems

Agriculture and Environment

Achievements and problems in the weed control in common wheat (Triticum Aestivum L.) and durum wheat (Triticum Durum Desf.)G. Delchev, M. Georgiev

Vokil – a new Bulgarian sunflower hybridG. Georgiev, V. Encheva, N. Nenova, Y. Encheva, D. Valkova, P. Peevska, G. Georgiev

Composition, digestibility, feeding value estimation of forage pea (Pisum sativum L.) varieties and hybrids in breeding process and genetic advance evaluationY. Naydenova, V. Kosev

Effect of age at first calving on conformation traits in Black-and-White cowsI. Marinov, Zh. Gergovska

Stimulating feeding and development of hypopharyngeal and thoracic glands of honeybees (Apis mellifera L.)R. Shumkova, I. Zhelyazkova

Histopathological changes in small intestines of broiler chickens with experimental aflatoxicosisN. Grozeva, I. Valchev, Ts. Hristov, L. Lazarov, Y. Nikolov

Histopathological pancreatic changes in broiler chickens with experimental aflatoxicosisI. Valchev, N. Grozeva, D. Kanakov, Y. Nikolov

Effect of foliar fertilization on sunflower (Helianhtus annuus L.)G. Milev

Changes in the productivity and the yield structural elements in some spring crops sown on damaged by frost crops of winter oilseed canola G. Delchev

Chromium level in water, sediment, aquatic plants and meat of common carp (Cyprinus carpio L.) from different water bodies in BulgariaE. Valkova, V. Atanasov, K. Velichkova, G. Kostadinova, M. Tzanova

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Visible – NIR reflectance for evaluation of luvisols and phaeozemsM. Todorova, M. Mihalache, L. Ilie, S. Atanassova

Groundwater quality as a source for irrigation in Strumica valley, Republic of MacedoniaB. Kovacevik, S. Mitrev, B. Boev, I. Karov, V. Zajkova Panova

Chemical and physical properties of the Albic luvisols from Albota-PiteștiM. Mihalache, L. Ilie, M. Todorova, N. Grozeva

Effects of salt-curing on physicochemical and technological properties of Japanese quail breast meat S. Ribarski, H. Lukanov, I. Penchev, A. Genchev, E. Lyutskanova

Study of physicochemical parameters of lamb meat from North East Bulgarian fine fleece breed of sheep and its crossbreds from internal breedingR. Slavov, G. Mihaylova, S. Ribarski, P. Slavova, D. Pamukova

Egg quality and shell colour characteristics of crosses between Araucana and Schijndelaar with highly productive White Leghorn and Rhode Island Red strainsH. Lukanov, A. Genchev, A. Pavlov

Influence of the sweet red papers extract on the quality and oxidative changes in fats of sausagesА. Kuzelov, V. Ilieva, N. Taskov, D. Saneva

Effect of dietary vitamin E supplementation on the oxidative stability of lipids and proteins in cooked porkT. Popova, P. Marinova, M. Ignatova

Product Quality and Safety

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Instruction for authors

Preparation of papersPapers shall be submitted at the editorial office typed on standard typing pages (A4, 30 lines per page, 62 characters per line). The editors recommend up to 15 pages for full research paper ( including abstract references, tables, figures and other appendices)The manuscript should be structured as follows: Title, Names of authors and affiliation address, Abstract, List of keywords, Introduction, Material and methods,Results, Discussion, Conclusion, Acknowledgements (if any), References, Tables, Figures.The title needs to be as concise and informative about the nature of research. It should be written with small letter /bold, 14/ without any abbreviations. Names and affiliation of authorsThe names of the authors should be presented from the initials of first names followed by the family names. The complete address and name of the institution should be stated next. The affiliation of authors are designated by different signs. For the author who is going to be corresponding by the editorial board and readers, an E-mail address and telephone number should be presented as footnote on the first page. Corresponding author is indicated with *.Abstract should be not more than 350 words. It should be clearly stated what new findings have been made in the course of research. Abbreviations and references to authors are inadmissible in the summary. It should be understandable without having read the paper and should be in one paragraph. Keywords: Up to maximum of 5 keywords should be selected not repeating the title but giving the essence of study. The introduction must answer the following questions: What is known and what is new on the studied issue? What necessitated the research problem, described in the paper? What is your hypothesis and goal ?Material and methods: The objects of research, organization of experiments, chemical analyses, statistical and other methods and conditions applied for the experiments should be described in detail. A criterion of sufficient information is to be possible for others to repeat the experi-ment in order to verify results.Results are presented in understandable

tables and figures, accompanied by the statistical parameters needed for the evaluation. Data from tables and figures should not be repeated in the text.Tables should be as simple and as few as possible. Each table should have its own explanatory title and to be typed on a separate page. They should be outside the main body of the text and an indication should be given where it should be inserted.Figures should be sharp with good contrast and rendition. Graphic materials should be preferred. Photographs to be appropriate for printing. Illustrations are supplied in colour as an exception after special agreement with the editorial board and possible payment of extra costs. The figures are to be each in a single file and their location should be given within the text. Discussion: The objective of this section is to indicate the scientific significance of the study. By comparing the results and conclusions of other scientists the contribution of the study for expanding or modifying existing knowledge is pointed out clearly and convincingly to the reader.Conclusion: The most important conse- quences for the science and practice resulting from the conducted research should be summarized in a few sentences. The conclusions shouldn't be numbered and no new paragraphs be used. Contributions are the core of conclusions. References:In the text, references should be cited as follows: single author: Sandberg (2002); two authors: Andersson and Georges (2004); more than two authors: Andersson et al.(2003). When several references are cited simultaneously, they should be ranked by chronological order e.g.: (Sandberg, 2002; Andersson et al., 2003; Andersson and Georges, 2004).References are arranged alphabetically by the name of the first author. If an author is cited more than once, first his individual publications are given ranked by year, then come publications with one co-author, two co-authors, etc. The names of authors, article and journal titles in the Cyrillic or alphabet different from Latin, should be transliterated into Latin and article titles should be translated into English. The original language of articles and books translated into English is indicated in parenthesis after the bibliographic reference (Bulgarian = Bg, Russian = Ru, Serbian = Sr, if in the Cyrillic, Mongolian =

Мо, Greek = Gr, Georgian = Geor., Japanese = Jа, Chinese = Ch, Arabic = Аr, etc.)The following order in the reference list is recommended:Journal articles: Author(s) surname and initials, year. Title. Full title of the journal, volume, pages. Example:Simm G, Lewis RM, Grundy B and Dingwall WS, 2002. Responses to selection for lean growth in sheep. Animal Science, 74, 39-50Books: Author(s) surname and initials, year. Title. Edition, name of publisher, place of publication. Example: Oldenbroek JK, 1999. Genebanks and the conservation of farm animal genetic resources, Second edition. DLO Institute for Animal Science and Heal th, Netherlands.Book chapter or conference proceedings: Author(s) surname and initials, year. Title. In: Title of the book or of the proceedings followed by the editor(s), volume, pages. Name of publisher, place of publication. Example: Mauff G, Pulverer G, Operkuch W, Hummel K and Hidden C, 1995. C3-variants and diverse phenotypes of unconverted and converted C3. In: Provides of the Biological Fluids (ed. H. Peters), vol. 22, 143-165, Pergamon Press. Oxford, UK.Todorov N and Mitev J, 1995. Effect of level of feeding during dry period, and body condition score on reproductive perfor-

thmance in dairy cows,IX International Conference on Production Diseases in Farm Animals, September 11–14, Berlin, Germany.Thesis:Hristova D, 2013. Investigation on genetic diversity in local sheep breeds using DNA markers. Thesis for PhD, Trakia University, Stara Zagora, Bulgaria, (Bg).

The Editorial Board of the Journal is not responsible for incorrect quotes of reference sources and the relevant violations of copyrights.

Animal welfareStudies performed on experimental animals should be carried out according to internationally recognized guidelines for animal welfare. That should be clearly described in the respective section “Material and methods”.

Volume 7, Number 3September 2015

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