Received: Revised: Accepted: June 14, 2014 August 07, 2014 March 06, 2015 Mannheimia haemolytica ... more Received: Revised: Accepted: June 14, 2014 August 07, 2014 March 06, 2015 Mannheimia haemolytica is the causative agent of several economically significant diseases of cattle and sheep, however, its role in causing infection in poultry is limited as secondary pathogen, co-infecting with viral pathogens like infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV) etc. The present study reports first time in this country regarding the isolation of M. haemolytica from adult commercial poultry flocks, initially reported with severe respiratory distress. Necropsy findings were quite similar to those found in Fowl cholera infection. The samples were also processed for the detection of any avian respiratory viruses in addition to testing for bacterial presence. The lab investigations led to the detection of M. haemolytica from the clinical samples and subsequent use of norfloxacin, selected on the basis of sensitivity pattern of the organism, resulted in curing of the ...
This study was designed to perform biological and molecular characterization of avian adenoviruse... more This study was designed to perform biological and molecular characterization of avian adenoviruses (AAVs) recovered from suspected cases of hydropericardium-hepatitis syndrome (HHS) in commercial poultry. Initially the samples were screened by Agar Gel Precipitation Test (AGPT) for the presence of AAVs followed by its confirmation and typing through polymerase chain reaction (PCR) focusing on already reported serotypes AAV-4, AAV-8 and AAV-10 elsewhere. These PCR-positive samples were further subjected to amplification of fiber gene, followed by conducting restriction fragment length polymorphism (RFLP) using restriction enzyme Alu. The selected isolates were further propagated through cell culture and pathogenic potential of selected isolates was determined by infecting chickens. In this study, out of a total 190 samples, 57.8% of suspected cases were found positive for AAV presence through AGPT while sub-type identification using PCR revealed 46.3% for these viruses belonging to A...
Received: Revised: Accepted: June 06, 2014 January 10, 2015 March 19, 2015 Due to the major occur... more Received: Revised: Accepted: June 06, 2014 January 10, 2015 March 19, 2015 Due to the major occurrence of infection by avian influenza virus (AIV) subtype H9N2 in broilers and broiler breeders at first 2-3 weeks of age in this country, where the virus is endemic since 1999, it has become necessary to devise new strategies for better protection of chicks at early age. In ovo vaccination is one of the approaches for providing neonatal resistance against various infectious diseases in chickens. The aim of this study was to develop in ovo vaccine against H9N2 serotype of AIV. For this purpose a vaccine strain of AIV H9N2 was used to develop both inactivated and live virus vaccines for experimentally inoculating18day old embryos. In addition to this two other groups of chicken were separately vaccinated with both inactivated oil-emulsion and live virus vaccines of H9N2 for comparison. The hatched chicks were monitored for the development of HI antibody response against AIV H9N2. All thes...
Received: Revised: Accepted: Online available: March 21, 2015 January 08, 2016 February 07, 2016 ... more Received: Revised: Accepted: Online available: March 21, 2015 January 08, 2016 February 07, 2016 June 29, 2016 The Live bird Markets (LBM) can serve as a paramount source of AIV infections. During routine Avian Influenza surveillance in Pakistan, low-pathogenic avian influenza virus subtype H4N6 was isolated first time from Khaki Campbell duck (Anas platyrhynchos) during 2010 and from broiler chicken during 2011 in the live bird markets (LBMs) from the port city of Karachi in Sindh province. Whole genome sequencing revealed introduction of a new reassortant Eurasian avian virus strain. Phylogenetically HA, NA, M, NP and PB2 genes clustered mostly with Russian strains of influenza viruses and PA gene with AI isolates from Netherlands, whereas NS and PB1 genes clustered with a Pakistani isolate of H3N1. Sequence analysis revealed a LP amino acid motif (PEKASR), avian-like receptors, conservation of amino acids at the receptor binding sites and the amino acids known to be associated wi...
Abstract The Escherichia coli (E. coli), Salmonella, Mycoplasma synoviae (MS) and Mycoplasma gall... more Abstract The Escherichia coli (E. coli), Salmonella, Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG) are the major avian bacterial pathogens closely related to avian colibacillosis, salmonellosis, and mycoplasmosis respectively. In present study a single-tube multiplex PCR (m-PCR) assay was developed to allow the rapid, specific and simultaneous detection of these pathogenic species of major avian bacterial diseases. For this purpose, species specific genes of the relevant pathogens were first identified and utilized to obtain similar thermal profile conditions in singleplex PCR against the species-specific target genes “vat” of E. coli, “FimH” of Salmonella, “DNA polymerase III PolC” of MS and “mgc2” gene of MG. The primer pairs amplified specifically DNA fragment of the target genes in singleplex PCR, of 200 bp (vat), 300 bp (fimH), 380 bp (DNA polymerase III PolC) and 450 bp (mgc2). The validated species-specific primers from the similar thermal profile of singleplex PCR were then simultaneously subjected to optimize multiplex PCR condition at Tm 63 °C and amplified the same size products on agarose gel. The m-PCR was able to detect and differentiate simultaneously the aforementioned four major pathogenic species. The assay was corroborated for validation against field trial at the laboratory. The developed multiplex PCR for simultaneous identification of the aforementioned avian bacterial pathogens will help to enhance the diagnostic procedures of different poultry laboratories for field samples.
Received: Revised: Accepted: June 14, 2014 August 07, 2014 March 06, 2015 Mannheimia haemolytica ... more Received: Revised: Accepted: June 14, 2014 August 07, 2014 March 06, 2015 Mannheimia haemolytica is the causative agent of several economically significant diseases of cattle and sheep, however, its role in causing infection in poultry is limited as secondary pathogen, co-infecting with viral pathogens like infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV) etc. The present study reports first time in this country regarding the isolation of M. haemolytica from adult commercial poultry flocks, initially reported with severe respiratory distress. Necropsy findings were quite similar to those found in Fowl cholera infection. The samples were also processed for the detection of any avian respiratory viruses in addition to testing for bacterial presence. The lab investigations led to the detection of M. haemolytica from the clinical samples and subsequent use of norfloxacin, selected on the basis of sensitivity pattern of the organism, resulted in curing of the ...
This study was designed to perform biological and molecular characterization of avian adenoviruse... more This study was designed to perform biological and molecular characterization of avian adenoviruses (AAVs) recovered from suspected cases of hydropericardium-hepatitis syndrome (HHS) in commercial poultry. Initially the samples were screened by Agar Gel Precipitation Test (AGPT) for the presence of AAVs followed by its confirmation and typing through polymerase chain reaction (PCR) focusing on already reported serotypes AAV-4, AAV-8 and AAV-10 elsewhere. These PCR-positive samples were further subjected to amplification of fiber gene, followed by conducting restriction fragment length polymorphism (RFLP) using restriction enzyme Alu. The selected isolates were further propagated through cell culture and pathogenic potential of selected isolates was determined by infecting chickens. In this study, out of a total 190 samples, 57.8% of suspected cases were found positive for AAV presence through AGPT while sub-type identification using PCR revealed 46.3% for these viruses belonging to A...
Received: Revised: Accepted: June 06, 2014 January 10, 2015 March 19, 2015 Due to the major occur... more Received: Revised: Accepted: June 06, 2014 January 10, 2015 March 19, 2015 Due to the major occurrence of infection by avian influenza virus (AIV) subtype H9N2 in broilers and broiler breeders at first 2-3 weeks of age in this country, where the virus is endemic since 1999, it has become necessary to devise new strategies for better protection of chicks at early age. In ovo vaccination is one of the approaches for providing neonatal resistance against various infectious diseases in chickens. The aim of this study was to develop in ovo vaccine against H9N2 serotype of AIV. For this purpose a vaccine strain of AIV H9N2 was used to develop both inactivated and live virus vaccines for experimentally inoculating18day old embryos. In addition to this two other groups of chicken were separately vaccinated with both inactivated oil-emulsion and live virus vaccines of H9N2 for comparison. The hatched chicks were monitored for the development of HI antibody response against AIV H9N2. All thes...
Received: Revised: Accepted: Online available: March 21, 2015 January 08, 2016 February 07, 2016 ... more Received: Revised: Accepted: Online available: March 21, 2015 January 08, 2016 February 07, 2016 June 29, 2016 The Live bird Markets (LBM) can serve as a paramount source of AIV infections. During routine Avian Influenza surveillance in Pakistan, low-pathogenic avian influenza virus subtype H4N6 was isolated first time from Khaki Campbell duck (Anas platyrhynchos) during 2010 and from broiler chicken during 2011 in the live bird markets (LBMs) from the port city of Karachi in Sindh province. Whole genome sequencing revealed introduction of a new reassortant Eurasian avian virus strain. Phylogenetically HA, NA, M, NP and PB2 genes clustered mostly with Russian strains of influenza viruses and PA gene with AI isolates from Netherlands, whereas NS and PB1 genes clustered with a Pakistani isolate of H3N1. Sequence analysis revealed a LP amino acid motif (PEKASR), avian-like receptors, conservation of amino acids at the receptor binding sites and the amino acids known to be associated wi...
Abstract The Escherichia coli (E. coli), Salmonella, Mycoplasma synoviae (MS) and Mycoplasma gall... more Abstract The Escherichia coli (E. coli), Salmonella, Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG) are the major avian bacterial pathogens closely related to avian colibacillosis, salmonellosis, and mycoplasmosis respectively. In present study a single-tube multiplex PCR (m-PCR) assay was developed to allow the rapid, specific and simultaneous detection of these pathogenic species of major avian bacterial diseases. For this purpose, species specific genes of the relevant pathogens were first identified and utilized to obtain similar thermal profile conditions in singleplex PCR against the species-specific target genes “vat” of E. coli, “FimH” of Salmonella, “DNA polymerase III PolC” of MS and “mgc2” gene of MG. The primer pairs amplified specifically DNA fragment of the target genes in singleplex PCR, of 200 bp (vat), 300 bp (fimH), 380 bp (DNA polymerase III PolC) and 450 bp (mgc2). The validated species-specific primers from the similar thermal profile of singleplex PCR were then simultaneously subjected to optimize multiplex PCR condition at Tm 63 °C and amplified the same size products on agarose gel. The m-PCR was able to detect and differentiate simultaneously the aforementioned four major pathogenic species. The assay was corroborated for validation against field trial at the laboratory. The developed multiplex PCR for simultaneous identification of the aforementioned avian bacterial pathogens will help to enhance the diagnostic procedures of different poultry laboratories for field samples.
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