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Clinical Observation on Mycobiota of Postmortem Samples of Commercial Broiler Suspected Forhemorrhagic Proventriculitis and Gizzard Erosion in Nepal

A new disease condition has emerged affecting replacement pullets. It affects the birds between 2 to 10 weeks of age. The mortality rate varies between 1 to 10% and ultimately the flock is left with creation of several small poorly feathered and pale chicks. Though it has not been possible to establish the exact cause of this condition, following three points has been taken in consideration. Hemorrhagic Proliferative Proventriculitis and Gizzard Erosion is an illness that involves one or more infectious agents.One the cause suspected for it is mycotic origion. The digestive tract and endocrine organs may be primary targets for these agents.The spectrum of signs or lesions are either caused by or exacerbated by nutritional, husbandry and hygienic factors.The response to the treatment is often poor but the symptomatic treatment with toxinbinder like toxolivum liquid,hepatocare, alongwith immunomodulaters like immunocare,and promin has significantly helped in ameliorating the mortality. Similar condition has been widely reported in broilers under various names like Malabsorption syndrome, infectious Proventriculitis, infectious runting syndrome, pale bird syndrome and stunting syndrome. Same condition is likely to be emmerging as a problem for coomercial broiler after 3rd week age.

Review of Litrature:

Mycotoxins T2 toxin produced by fusarium is a caustic irritant. It causes necrosis of mucosa of proventriculus, gizzard and feather epithelium. Citrinin which is basically a nephrotoxin can also cause fissures in the gizzard Oosporein – In oosporein poisoning one may notice that the proventriculus has enlarged circumference at the isthumus and the mucosa is covered with pseudomembranous exudate (necrosis may occur at the isthumus).Cyclopiazonic acid (CPA) – Lesions occur in proventriculus, gizzard, liver and spleen. Theproventriculus is dialated and the mucosa is thickened by hyperplasia and ulceration. Mucosal necrosis may occur in gizzard (Dr Manuel Contreras and Dr Douglas Zavieso).

Infectious Factors Adenovirus – According to case reports, gizzard erosion is characterised by adenovirus intranuclear inclusion bodies in epithelial cells. Reovirus infection could be a factor in the pathogenesis of histamine associated proventricular enlargement High levels of Dietory biogenic amine’s like histamine, 3HT, 5HT,histidine, dopamine, gizzerosine and serotonine, can be found in dietary constituents such as tankage fish meal, corn screening, soyabean meal, vitamin premixes, fats, poultry meal, meat and bone meal. The biogenic amines are decarboxylation breakdown products of amino acid catabolism and these amines are considered toxic to animals. The potential for biogenic amine build-up is real in animal by product meal and is the result of breakdown of the product. Histamine is produced in the poultry feed under proper temperature and moisture conditions by microbial decarboxylation of histidine. Reduced growth, poor feathering and proventricular enlargement have been associated with histamine toxicity in chickens. Histamine toxin problem in chicken generally has been associated with the intake of fish meal which contain high level of histidine Gizzerosine is a compound found in overheated fish meal due to interaction of caesine with histidine and acts as a factor causing gizzard erosion or ulceration in chicks. It can also be formed if the temperature of fish meal increases by incorrect handling during transportation or storage particularly in hot weather( Martin D. Ficken,) .

Then gizzerosine concentration of burnt fish meal would be low because of degradation of protein. However it is also likely that fish meal with good colour, odour, taste and physical properties may contain a large enough quantity of gizzerosine.Gizzerosine stimulates proventricular gland secretarycells to release excessive hydrochloric acid. Gizzard lesions result from the runway digestive effects of hyperacidity. Opportunistic bacteria may subsequently colonise the nutrient rich biodetrius. The cells of the glandular alveoli of the proventriculus secrete hydrochloric acids and pepsinogen (pepsin) which is a digestive enzyme required for initial digestion of proteins. Any lesion in the preventricular glands will interfere with the secretion of pepsin with subsequent impairment of protein digestion and utilisation. The results are poor production performance, unthriftiness and poor feed conversion. This could also explain why some of the birds affected with this condition pass undigested or poorly digested feed in faeces (G. D. Butcher) .

. A Reo virus strain (SS 412 stain) was isolated from an outbreak of proventriculitis/malabsorption of syndrome; the role of this reovirus strain in that outbreak was proven in experimental studies. Further studies have demonstrated that chicks from breeder hens which have been vaccinated with oil imulsion SS 412 virus vaccine were protected against. Proventriculitis following experimental challenge with the SS 412 reovirus strain. Anaerobic bacteria like clostridia are sometimes found as secondary invaders resulting in either ulcerative enteritis or necrotic hepatitis. It will be interesting to study the effect of viruses also since both of them cause the lesions in the proventriculus (G. D. Butcher).Other Factors Rather than ameliorating the effects of malabsorbtion syndrome, vitamin A caused a further reduction in body weight and bone ash according to a study. Supplementation of vitamin E significantly reduced both mortality and the effects of disease in body weight gain in an outbreak of pale bird syndrome in broiler chicks of 3 weeks. Amino acid imbalance (lysine and methoinine especially), excess dietary copper sulfate, lack of dietary fibre, deprivation of food and water have also been found to be responsible for the heamorhagic prolifirative proventriculitis and gizzard erosion. Lesions Proventriculi lose their normal flusiform shape and normal constriction at the junction with gizzard are diffusely enlarged and have a thickened and turgidwall. Thickening of the wall is more marked upon incising the proventriculus.The proventricular glands protrude irregularly from the mucosal surface, lose their normal pattern and contain milky fluid that could be expressed with slight pressure. The gizzard is often smaller than normal and flabby. The gizzard peels off easily with haemorrhagic ulceration of the gizzard wall(Dr. Avinash Dhawale) . An clinical epidemiological investigation of Acute sudden death syndrome due to which 31 mules from a herd of 9oo died within the period of 2006-7-12 to2006-10-21 in Udayapur District Nepal. These animals were being used for good transportation work in hilly region of Nepal. On rout these animals were being fed only whole maize and grame.Normally healthy looking animals started dying suddenly. Initially suspected for acute bacterial disease and treated with broad-spectrum antibiotics and vaccinated with bacterial vaccine.On close observation of herd their feed stuff revealed grains fed to these animal during rainy season was found 20% moldy in appearance. On Postmortem examination acute severe congestion and hemorrhages in liver, lung, spleen heart, intestinal mucosa were found.Histopathological examination of tissue from these organs revealed infiltration of mononuclear cell in tissue indicative of chronic nature of condition.Continous use of apparently 15-20% moldy grain (maize, gram) was used as feed. On laboratory analysis of sample of same grain was found to be containing 60-110CFU/gm of Penicillium spp of fungus ( Dr.Karki et.al http://www.alumbo.com/article/43263. Feed contamination can lead to nutrient losses and detrimental effects on animal health and production. Feed mould counts ranged from

Materialand Methodes

Observation of flock

Flock size
























lesions observed during postmorten

• Anemic appearance, poor growth, pale skin

• Mucous in URT.

• Distorted shape of proventriculus,gizzard.

• Swollen large ,small intestine.

• Liver pale, enlarged with white spate.

• Enlarged spleen.

• Emphymatouse lung.

• Ascites.

• Milky fluid oozing from Proventricular gland.

• Hemorrhage in Proventricular muscle.

• Yellow tinge with white ulceration in gizzard.

Mycology Laboratory Examination






Isolate fungus







Penicillium spp.

Candida spp.



Treatment adviced

Broadespectrum antibiotics,Liquide toxinbider like toxolivum,Hepatocare, Immunomodulaters like immunocare,and promin for a week.

Result and Discussion: Mcological screening of postmortem samples of liver,spleen,proventriculus of bird revealed the presenceFungus spp like Penicillium,Aspergillus in 194 samples out of 284 samples were cultured in potato dextrose agar medium during the period 2007-7-1 to 2008-3-1.The postmortem lesions and age of flock had the similarities as reported by other workers.As there has not yet any observation of mycobiota of poultry lesion carried out this finding serves as a preliminary information inthis regard need to be further validated.As modern poultry feed milling operation toxinbinder is being used but still it is not going to suffient to combat the effect of mycotoxin produced by storage fungus.This finding indicates that after 3rd week of age brioler need to be suplimented with liquid toxinbinder to reduce the loss from this emmerging disease syndrome need to be looked into.


1;Cause of gizzard erosion and Proventriculitis in broilers. Dr Manuel Contreras and Dr Douglas Zavieso Poultry International July 2006 pp16-20 Cyclopiazonic acid production by Aspergillus flavus and its effects on broiler chickens. J W Dorner, R J Cole, L G Lomax, H S Gosser, and U L Diener Appl Environ Microbiol. 1983 September; 46(3): 698–703.

2:DISEASES OF POULTRY: Martin D. Ficken, D.V.M., Ph.D., Dip. A.C.V.P., A.C.P.V. College of Veterinary Medicine North Carolina State Universit Raleigh, North Carolina

3:Feed Passage in Broilers – A Complex Problem G. D. Butcher, DVM, Ph.D., A. H. Nilipour, Ph.D., R. D. Miles, Ph.D. University of Florida College of Veterinary Medicine, Gainesville, FL., Amir H. Nilipour, PhD, Director of Investigation and Quality Assurance, Grupo Melo, S.A., Panama, Republic of Panama .

4:http://www.wattnet.com/Archives/Docs/701pi36.pdf?CFID=25710&CFTOKEN=74030876 gizzard erosion Proventriculitis poultry A new disease condition has emerged affecting replacement pullets in India. — By Dr. Avinash Dhawale .

5:The Mycobiota and Toxicity of Equine Feeds:Journal Veterinary Research Communication volume 31,number 8 November 2007: K. M. Keller1, B. D. Queiroz1, L. A. M. Keller1, J. M. M. Ribeiro1, L. R. Cavaglieri2 , M. L. González Pereyra2, A. M. Dalcero2 and C. A. R. Rosa1(1) Departamento de Microbiologia e Imunologia Veterinária, Universidade Federal Rural do Rio de Janeiro, Instituto de Veterinária, Rio de Janeiro, Brazil 2;Departamento de Microbiología e Inmunología, Universidad Nacional de Río Cuarto, Río Cuarto, Argentina Accepted: 28 August 2006 Published online: 6 February 2007

6: Unacceptably High Fungal Levels Identified in Horse Feeds by: Stacey Oke, DVM, MSc, September 08 2007, Article # 10366 The Horse.com

7: Clinical-Epidemiological Investigation of Moldy Corn Poisoning in mules due to Penicillium spp and its treatment by using toxinbinder, herbal liver tonic.immunomodulater at Udayapur District, Nepal 1: Dr.Kedar Karki Vet. Officer,Dr.Poornima ManandharSVO Central Vet. Laboratory. Tripureswor Kathmandu Nepal http://www.alumbo.com/article/43263

About the Author

Senior Vet.Officer,Central Veterinary Laboratory Kathmandu Nepal M.V.St. Preventive Veterinary Mrdicine

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