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Incidence of Fowl Cholera in Desi Fowl Farms of Tirupur District

V. S. Vadivoo M. Arthanari Eswaran T. Hariharan
Vol 8(8), 105-109
DOI- http://dx.doi.org/10.5455/ijlr.20171116111150

Desi birds are often susceptible to fowl cholera, a serious, highly contagious disease caused by the bacterium, Pasteurella multocida. This study involves the diagnosis of fowl cholera by gross necropsy examination, isolation and identification of organism by culture and biochemical method. During the span of 8 months (from January 2017 to August 2017) six different flocks of adult desi chicken (with an average flock size of 70 birds per flock) were subjected to necropsy examination and a total of 30 carcasses were examined for diagnosis of the disease. Fowl cholera was confirmed in all six flocks by gross and microscopic examination and bacterial isolation by culture method and biochemical method. The women self help groups were advised to follow strict biosecurity and ethno veterinary practices for prevention and control of diseases.


Keywords : Confirmation Fowl Cholera Microscopic Examination Necropsy Treatment

Tirupur district has many women self-groups sponsored by the state government. Among the various programmes offered to the women self-help groups, backyard desi fowl farming is one of the major revenue generating avenue. Small flock of around 40 to 100 desi birds is normally reared in the backyard. The major disease outbreaks among these birds are Ranikhet disease, fowl cholera and fowl pox.

Pasteurella multocida, a gram-negative bacterium, is the causative agent of fowl cholera in birds.  Among avian strains of P. multocida, serogroup A strains cause the majority of fowl cholera. This organism being an opportunistic pathogen is also a common commensal in the upper respiratory tract of birds. The infection leads to acute septicaemia to chronic and localised infections and the morbidity and mortality might be up to 100 per cent as the pathogen resists phagocytosis after invading the internal tissues and multiply very quickly. The route of infection is oral or nasal with transmission via nasal exudate, faeces, contaminated soil, equipment, and people. The incubation period is usually 5-8 days. (Paul Mcmullin 2004).

Fowl cholera does not have any zoonotic implication when exposed by the oral or subcutaneous routes. Generally clinical signs of fowl cholera may be similar to salmonellosis, colibacillosis and listeriosis in poultry. Differentiation is based on isolation and identification, as P. multocida can be readily cultured from cases of fowl cholera. Since the routine nature of isolation and identification of P. multocida by culture method and biochemical tests have been used widely without dubious results, confirmation of the results is acceptable (OIE, 2012). Identification of P. multocida is based on the results of biochemical tests, which include carbohydrate fermentation, enzyme production, and selected metabolite production.

Materials and Methods

A total of 30 carcasses from six different backyard poultry farms from various areas of Tirupur district were examined. Few live ailing birds (brought along with the carcass) exhibited the signs of dullness, off feed, paleness in comb along with whitish diarrhoea. Heart blood smears, aspirates of heart blood and impression smears of spleen, liver and lung were collected from dead desi fowls and stained with Leishman stain (Akhtar. 2013). The heart blood was also inoculated in Brain Heart Infusion (BHI) broth and incubated at 370C overnight and the broth culture was streaked onto blood agar and MacConkey agar. The colonies suggestive of P. multocida were subjected to biochemical tests for identification. The biochemical tests included IMVIC, sugar fermentation test and catalase and oxidase test.

Antibiotic sensitive assay was performed for the P.multocida isolates as per the disc diffusion method of Kirby-Bauer (Bauer et al., 1966).

Results and Discussion

The commonly observed external signs in the live birds were anorexia, ruffled feathers, oral and nasal mucus discharge, cyanosis and white diarrhoea.

On post-mortem the lesions observed were general septicaemia like vascular disturbances, showing general passive hyperemic congestion throughout the carcass. Petechial and ecchymotic haemorrhages were seen in the abdominal region and haemorrhages were found in the intestinal mucosae (Fig. 1a) and on subserosal surfaces in the thoracic and abdominal cavities.  The liver and spleen were swollen with typical multiple small focal areas of coagulative necrosis (Fig. 2a and 2 b). Severe congestion and haemorrhages were noticed on the heart and lungs (Fig. 3a and 3b). These observations were similar as reported by Srinivasan et al. (2011) and Aravinth et al. (2016) in broiler chicks.

Fig .1a: Haemorrhages in intestinal mucosae

Fig. 2a and 2b: Liver showing typical multiple small focal areas of coagulative necrosis

Fig. 3a and 3b: Heart and lungs showing haemorrhages and congestion

Heart blood smear, liver and spleen impression smears showed characteristic bipolar organisms on Leishman staining (Fig. 4) and Gram negative coccobacilli by Gram’s staining method were in accordance with Ievy et al. (2013) and Akhtar et al. (2013). The isolates showed typical cultural characteristics of dew drop, mucoid, non haemolytic colonies in blood agar (Fig. 5). No growth was observed in MacConkey agar. Gram’s staining of the smears revealed characteristic gram- negative coccobacillary organisms. The isolates were positive for indole, nitrate reduction, oxidase and catalase and negative for methyl red and Voges–Proskauer tests. The carbohydrates fermented were glucose, mannose, galactose, fructose, sucrose and manitol and the carbohydrates that were not fermented included rhamnose, cellobiose, raffinose, inulin, erythritol, adonitol, m-inositol, and salicin. These results were same as reported in OIE (2012) and  Panna  et al. (2015)

bipolar organism

 

 

Fig. 4 : Leishman staining

 

 

Mucoid Colony

 

Fig. 5: Dew drop, mucoid, non haemolytic colonies in blood agar

Antibiotic sensitivity test revealed that the isolate was sensitive to Gentamicin, Amikacin Doxycycline, Ceftriaxone and Chlortetracycline. Resistance was observed to antibiotics such as Co-trimoxazole, Ciprofloxacin, Enrofloxacin and Furozolidone. Similar sensitivity pattern was observed by Aravinth et al. (2016). Water sample collected from the farm premises, tested for coliforms revealed 7×10 2 CFU/mL. The ideal coliform count should be 0 cfu/ml and a level above this indicates contamination which might also add to the complexity of the disease and multiple bacterial infection generally reduces the immune system. As per the antibiotic sensitivity test results the flock with high mortality was treated with Gentamicin and the recovery was observed after 5 days. The aminoglycosides remain in the digestive tract so are effective in the treatment of enteric infections. The bacterium is easily destroyed by environmental factors and disinfectants, but may persist for prolonged periods in soil. This could probably be due to poor disinfection procedure or improper hygienic measures followed for disinfecting the farms.  Reservoirs of infection might be present in other species such as rodents, cats, and possibly pigs. This was confirmed by the statistical study reported by Tahir et al. (2003) which says that farms where previous outbreaks of fowl cholera favored the onset in commencing flocks. Predisposing factors include high density and concurrent infections such as respiratory viruses. Hence the women self-help groups were advised to follow strict biosecurity and ethnoveterinary practices for prevention and control of diseases.

References

  1. Akhtar, M. 2013. Isolation, identification and characterization of Pasteurella multocida from chicken and development of oil based vaccine, MS thesis, Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh
  2. Aravinth A, Prakash S and Hariharan P. 2016. Incidence of Fowl Cholera with Anomalous Lesions in Laying Hens: A Case Study. British Journal of Poultry Sciences 5 (1): 09-12
  3. Bauer AW, Kirby WMM, Sherris JC and Turck M. 1966. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 45:493-496.
  4. Ievy S, Khan MRF, Islam MA, Rahman MB 2013. Isolation and identification of Pasteurella multocida from chicken for the preparation of oil adjuvanted vaccine. Bangladesh Journal of Veterinary Medicine, 2: 1-4.
  5. Office International des epizooties. 2012. Manuals of Standards for Diagnostic Test and Vaccine. 7th edition, France, 19 (2), 626-637.
  6. Panna SN, Nazir NH,  Bahanur Rahman M,  Ahamed S,  Saroare G,  Chakma S, Tazrin K and  Majumder UH, 2015. Isolation and molecular detection of Pasteurella multocida Type A from naturally infected chickens, and their histopathological evaluation in artificially iTirupur district has many women self-groups sponsored by the state government. Among the various programmes offered to the women self-help groups, backyard desi fowl farming is one of the major revenue generating avenue. Small flock of around 40 to 100 desi birds is normally reared in the backyard. The major disease outbreaks among these birds are Ranikhet disease, fowl cholera and fowl pox.

    Pasteurella multocida, a gram-negative bacterium, is the causative agent of fowl cholera in birds.  Among avian strains of P. multocida, serogroup A strains cause the majority of fowl cholera. This organism being an opportunistic pathogen is also a common commensal in the upper respiratory tract of birds. The infection leads to acute septicaemia to chronic and localised infections and the morbidity and mortality might be up to 100 per cent as the pathogen resists phagocytosis after invading the internal tissues and multiply very quickly. The route of infection is oral or nasal with transmission via nasal exudate, faeces, contaminated soil, equipment, and people. The incubation period is usually 5-8 days. (Paul Mcmullin 2004).

    Fowl cholera does not have any zoonotic implication when exposed by the oral or subcutaneous routes. Generally clinical signs of fowl cholera may be similar to salmonellosis, colibacillosis and listeriosis in poultry. Differentiation is based on isolation and identification, as P. multocida can be readily cultured from cases of fowl cholera. Since the routine nature of isolation and identification of P. multocida by culture method and biochemical tests have been used widely without dubious results, confirmation of the results is acceptable (OIE, 2012). Identification of P. multocida is based on the results of biochemical tests, which include carbohydrate fermentation, enzyme production, and selected metabolite production.

    Materials and Methods

    A total of 30 carcasses from six different backyard poultry farms from various areas of Tirupur district were examined. Few live ailing birds (brought along with the carcass) exhibited the signs of dullness, off feed, paleness in comb along with whitish diarrhoea. Heart blood smears, aspirates of heart blood and impression smears of spleen, liver and lung were collected from dead desi fowls and stained with Leishman stain (Akhtar. 2013). The heart blood was also inoculated in Brain Heart Infusion (BHI) broth and incubated at 370C overnight and the broth culture was streaked onto blood agar and MacConkey agar. The colonies suggestive of P. multocida were subjected to biochemical tests for identification. The biochemical tests included IMVIC, sugar fermentation test and catalase and oxidase test.

    Antibiotic sensitive assay was performed for the P.multocida isolates as per the disc diffusion method of Kirby-Bauer (Bauer et al., 1966).

    Results and Discussion

    The commonly observed external signs in the live birds were anorexia, ruffled feathers, oral and nasal mucus discharge, cyanosis and white diarrhoea.

    On post-mortem the lesions observed were general septicaemia like vascular disturbances, showing general passive hyperemic congestion throughout the carcass. Petechial and ecchymotic haemorrhages were seen in the abdominal region and haemorrhages were found in the intestinal mucosae (Fig. 1a) and on subserosal surfaces in the thoracic and abdominal cavities.  The liver and spleen were swollen with typical multiple small focal areas of coagulative necrosis (Fig. 2a and 2 b). Severe congestion and haemorrhages were noticed on the heart and lungs (Fig. 3a and 3b). These observations were similar as reported by Srinivasan et al. (2011) and Aravinth et al. (2016) in broiler chicks.

    Fig .1a: Haemorrhages in intestinal mucosae

    Fig. 2a and 2b: Liver showing typical multiple small focal areas of coagulative necrosis

    Fig. 3a and 3b: Heart and lungs showing haemorrhages and congestion

    Heart blood smear, liver and spleen impression smears showed characteristic bipolar organisms on Leishman staining (Fig. 4) and Gram negative coccobacilli by Gram’s staining method were in accordance with Ievy et al. (2013) and Akhtar et al. (2013). The isolates showed typical cultural characteristics of dew drop, mucoid, non haemolytic colonies in blood agar (Fig. 5). No growth was observed in MacConkey agar. Gram’s staining of the smears revealed characteristic gram- negative coccobacillary organisms. The isolates were positive for indole, nitrate reduction, oxidase and catalase and negative for methyl red and Voges–Proskauer tests. The carbohydrates fermented were glucose, mannose, galactose, fructose, sucrose and manitol and the carbohydrates that were not fermented included rhamnose, cellobiose, raffinose, inulin, erythritol, adonitol, m-inositol, and salicin. These results were same as reported in OIE (2012) and  Panna  et al. (2015)

    bipolar organism

     

    Fig. 4 : Leishman staining

     

     

    Mucoid Colony

     

    Fig. 5: Dew drop, mucoid, non haemolytic colonies in blood agar

    Antibiotic sensitivity test revealed that the isolate was sensitive to Gentamicin, Amikacin Doxycycline, Ceftriaxone and Chlortetracycline. Resistance was observed to antibiotics such as Co-trimoxazole, Ciprofloxacin, Enrofloxacin and Furozolidone. Similar sensitivity pattern was observed by Aravinth et al. (2016). Water sample collected from the farm premises, tested for coliforms revealed 7×10 2 CFU/mL. The ideal coliform count should be 0 cfu/ml and a level above this indicates contamination which might also add to the complexity of the disease and multiple bacterial infection generally reduces the immune system. As per the antibiotic sensitivity test results the flock with high mortality was treated with Gentamicin and the recovery was observed after 5 days. The aminoglycosides remain in the digestive tract so are effective in the treatment of enteric infections. The bacterium is easily destroyed by environmental factors and disinfectants, but may persist for prolonged periods in soil. This could probably be due to poor disinfection procedure or improper hygienic measures followed for disinfecting the farms.  Reservoirs of infection might be present in other species such as rodents, cats, and possibly pigs. This was confirmed by the statistical study reported by Tahir et al. (2003) which says that farms where previous outbreaks of fowl cholera favored the onset in commencing flocks. Predisposing factors include high density and concurrent infections such as respiratory viruses. Hence the women self-help groups were advised to follow strict biosecurity and ethnoveterinary practices for prevention and control of diseases.

    References

    1. Akhtar, M. 2013. Isolation, identification and characterization of Pasteurella multocida from chicken and development of oil based vaccine, MS thesis, Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh
    2. Aravinth A, Prakash S and Hariharan P. 2016. Incidence of Fowl Cholera with Anomalous Lesions in Laying Hens: A Case Study. British Journal of Poultry Sciences 5 (1): 09-12
    3. Bauer AW, Kirby WMM, Sherris JC and Turck M. 1966. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 45:493-496.
    4. Ievy S, Khan MRF, Islam MA, Rahman MB 2013. Isolation and identification of Pasteurella multocida from chicken for the preparation of oil adjuvanted vaccine. Bangladesh Journal of Veterinary Medicine, 2: 1-4.
    5. Office International des epizooties. 2012. Manuals of Standards for Diagnostic Test and Vaccine. 7th edition, France, 19 (2), 626-637.
    6. Panna SN, Nazir NH,  Bahanur Rahman M,  Ahamed S,  Saroare G,  Chakma S, Tazrin K and  Majumder UH, 2015. Isolation and molecular detection of Pasteurella multocida Type A from naturally infected chickens, and their histopathological evaluation in artificially infected chickens in Bangladesh. Journal of Advanced Veterinary and Animal Research. 2(3): 338-345.
    7. Paul McMullin 2004. A Pocket Guide to: Poultry Health and Disease, Chapter 6 pg 86-92
    8. Srinivasan P , Gopalakrishnamurthy, TR, Mohan B and Saravanan S. 2011 Occurrence Of Sub Acute Fowl Cholera In A Broiler Flock. Tamilnadu Journal of Veterinary and Animal Sciences, 7 (1): 45-47.
    9. Tahir B, Durrani, FR, Farooq M. Durrani, Z, Sar Zamin;. Khan, MA, and Riaz A.  2003 Prevalence of Fowl Cholera(Pasteurella multocida) in Commercial Broiler Breeder. Flocks Maintained in Abbottabad and Mansehra. Journal of Animal and Veterinary Advances, 2(8):444-447.

     

    nfected chickens in Bangladesh. Journal of Advanced Veterinary and Animal Research. 2(3): 338-345.

  7. Paul McMullin 2004. A Pocket Guide to: Poultry Health and Disease, Chapter 6 pg 86-92
  8. Srinivasan P , Gopalakrishnamurthy, TR, Mohan B and Saravanan S. 2011 Occurrence Of Sub Acute Fowl Cholera In A Broiler Flock. Tamilnadu Journal of Veterinary and Animal Sciences, 7 (1): 45-47.
  9. Tahir B, Durrani, FR, Farooq M. Durrani, Z, Sar Zamin;. Khan, MA, and Riaz A.  2003 Prevalence of Fowl Cholera(Pasteurella multocida) in Commercial Broiler Breeder. Flocks Maintained in Abbottabad and Mansehra. Journal of Animal and Veterinary Advances, 2(8):444-447.

 

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