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Clinico-Pathological And Pathomorphological Studies On Co-Infection Of Avian Influenza (H9n2) With Escherichia Coli In Broiler Chicken

By: Shahid Jaleel (2003-VA-93) | Dr. Muhammad Asif Idrees.
Contributor(s): Prof. Dr. Muhammad Younus | Dr. Muhammad Arshad.
Material type: materialTypeLabelBookPublisher: 2016Description: 65p.Subject(s): Department of PathologyDDC classification: 2552-T Dissertation note: E.coli is an important pathogen of domestic poultry and is prevalent in commercial poultry. LPAIV H9N2 infections are emerging respiratory problems in poultry industry, causing huge economic losses especially in the presence of other co-infecting pathogens such as E.coli. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. The aim of the present study was to investigate the infection of LPAIV A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) in chickens challenged with E.coli (O78:K80). This study had three objectives. First, it is designed to develop co-infection experimental models LPAIV (H9N2) + bacteria (E.coli) in the avian model. Second, it aims to study the hematological and biochemical alterations during co-infection in avian model. Finally to study the pathological and histological alterations during co-infection in avian model, this study will help researchers and veterinarians in implementation of necessary control measures. E.coli stockculture was prepared by inoculating MacConkey’s agar with a loop full of reference E.coli strain culture and incubating at 37°C for 24 h. The estimated colony count was confirmed by plating 0.1 ml of a 104 and a 105 dilution of the final culture onto separate MacConkey’s agar plates. Avian influenza A virus, A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) was obtained from Poultry Research Institute (PRI) Rawalpindi Pakistan. Viral stocks were prepared and titrated in 9-day-old to 10-day-old chicken embryonated eggs the median embryo infectious dose (EID50) was computed using previously reported approaches The viral stocks were diluted in medium containing antimicrobials to give a final titre of 106 EID50/ ml The study were ran on 80 broiler chicks (3week old), procured from local hatchery. All fowl were held serologically innocent and free from flu virus by haemagglutination inhibition (HI). Chicken were infected under experimental conditions with E.coli (O78:K80) and low pathogenic avian influenza (LPAI) strain (A/chicken/Pakistan/UDL-01/08) (H9N2) alone or in combination. The experimental groups were identified as follows: negative control, E.coli, AI, and E.coli plus AI. Infected birds showed clinical signs of differing severity, with the most prominent disease signs appearing in birds of the E.coli plus AI group. Moreover, birds in E.coli plus AI group showed significant decrease in weight, enhanced macroscopic and microscopic pathological lesions. Specifically, the survival rate was 60%, 90%, and 100% in birds inoculated with E.coli + AI, E.coli and control negative or AI virus alone, respectively. Hematological studies revealed anemia, thrombocytopenia and leukopenia especially in co-infected birds. Biochemical studies revealed a significant decrease in total protein, glucose and albumin concentration with significant increase of activities of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Prominent increase in creatinine, urea and uric acid were significantly detected in the infected chicken. The results showed that experimental co-infection of E.coli and H9N2 increased the severity of clinical signs, mortality rate and gross lesions and suggest than E.coli infection can induce higher economic losses and mortality if H9N2 LPAIV is also present. The HI titer against LPAIV infection in the co-infected group was significantly higher than the HI titer of AI group, which may indicate that E.coli could promote the propagation of H9N2 LPAIV or stimulate the immune response. The present study revealed that co-infection E.coli and H9N2 LPAIV caused more serious synergistic pathogenic effects and indicates the role of both pathogens as complicating factors in poultry infections.  
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Item type Current location Collection Call number Status Date due Barcode Item holds
Thesis Thesis UVAS Library
Thesis Section
Veterinary Science 2552-T (Browse shelf) Available 2552-T
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E.coli is an important pathogen of domestic poultry and is prevalent in commercial poultry. LPAIV H9N2 infections are emerging respiratory problems in poultry industry, causing huge economic losses especially in the presence of other co-infecting pathogens such as E.coli. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. The aim of the present study was to investigate the infection of LPAIV A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) in chickens challenged with E.coli (O78:K80).
This study had three objectives. First, it is designed to develop co-infection experimental models LPAIV (H9N2) + bacteria (E.coli) in the avian model. Second, it aims to study the hematological and biochemical alterations during co-infection in avian model. Finally to study the pathological and histological alterations during co-infection in avian model, this study will help researchers and veterinarians in implementation of necessary control measures. E.coli stockculture was prepared by inoculating MacConkey’s agar with a loop full of reference E.coli strain culture and incubating at 37°C for 24 h. The estimated colony count was confirmed by plating 0.1 ml of a 104 and a 105 dilution of the final culture onto separate MacConkey’s agar plates. Avian influenza A virus, A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) was obtained from Poultry Research Institute (PRI) Rawalpindi Pakistan.
Viral stocks were prepared and titrated in 9-day-old to 10-day-old chicken embryonated eggs the median embryo infectious dose (EID50) was computed using previously reported approaches The viral stocks were diluted in medium containing antimicrobials to give a final titre of 106 EID50/ ml The study were ran on 80 broiler chicks (3week old), procured from local hatchery. All fowl were held serologically innocent and free from flu virus by haemagglutination inhibition (HI).
Chicken were infected under experimental conditions with E.coli (O78:K80) and low pathogenic avian influenza (LPAI) strain (A/chicken/Pakistan/UDL-01/08) (H9N2) alone or in combination. The experimental groups were identified as follows: negative control, E.coli, AI, and E.coli plus AI. Infected birds showed clinical signs of differing severity, with the most prominent disease signs appearing in birds of the E.coli plus AI group. Moreover, birds in E.coli plus AI group showed significant decrease in weight, enhanced macroscopic and microscopic pathological lesions. Specifically, the survival rate was 60%, 90%, and 100% in birds inoculated with E.coli + AI, E.coli and control negative or AI virus alone, respectively. Hematological studies revealed anemia, thrombocytopenia and leukopenia especially in co-infected birds. Biochemical studies revealed a significant decrease in total protein, glucose and albumin concentration with significant increase of activities of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Prominent increase in creatinine, urea and uric acid were significantly detected in the infected chicken. The results showed that experimental co-infection of E.coli and H9N2 increased the severity of clinical signs, mortality rate and gross lesions and suggest than E.coli infection can induce higher economic losses and mortality if H9N2 LPAIV is also present. The HI titer against LPAIV infection in the co-infected group was significantly higher than the HI titer of AI group, which may indicate that E.coli could promote the propagation of H9N2 LPAIV or stimulate the immune response. The present study revealed that co-infection E.coli and H9N2 LPAIV caused more serious synergistic pathogenic effects and indicates the role of both pathogens as complicating factors in poultry infections.


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