Introduction: Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals still remains a challenge, as isolation and identification of A. pleuropneumoniae serotypes is difficult and quantification of the bacteria on agar plates is often almost impossible. The aim of the study was to develop and evaluate a serotype-specific quantitative TaqMan probe-based PCR for detection of serotype 2 in pig lungs, tonsils, and nasal swabs.

Introduction: The study objective was to develop and evaluate a new TaqMan multiplex real-time PCR method for Salmonella, L. monocytogenes, and verotoxigenic Escherichia coli (VTEC) detection in slaughtered animal carcasses.

In this study five serotypes of E. coli were isolated from chickens and identified as O1, O2, O6, O78 and O126 out of 33 isolates derived from a total of 60 samples. SDS-PAGE revealed that four proteins were characteristic and shared in all these serotypes at the molecular weight of 21, 30, 55 and 74 kDa of which 55 and 74 kDa proteins were fully reacted with the antisera against E. coli in the western blot. Other proteins are present but varied from one serotype to another.

Molecular detection and differentiation of Pasteurella multocida strain involved in a separate fowl cholera outbreak in a turkey flock farm located in El-Menofia Governorate, Egypt early 2008 was investigated. The isolated strain was compared with an Egyptian Pasteurella multocida isolate that was previously isolated from turkey flock during last decade besides four vaccinal strain (A:5, A:8, A:9 and D:2) on phenotypic and genotypic characterization basis. Phenotypically all the strains were similar as all the strains produce non haemolytic colonies on blood agar, and all the strains revealed similar biochemial behaviour. On the other hand, the genomic typing of all the stains using rep-PCR techniques [repetitive BOX elements, enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic (REP) polymerase chain reaction (PCR)] differentiated the six Pasteurella multocida strains into six different profiles. The molecular identity between the Pasteurella multocida 2008 strain and the previously isolated strain was 76.6 % and were ranged from 65.2% to 79.2% with the 4 vaccinal strains. These results reported the continuous mutations of the field Pasteurella multocida strains among poultry flocks in Egypt indicating the urgent need for the frequent and continuous molecular epidemiological investigations of fowl cholera outbreaks in various poultry flocks to detect these new strains and update the fowl cholera vaccines.

The present study was aimed to investigate the gynaeco-pathological disorders by post-mortem and histopathological examination, and to identify the associated bacteria. A total of 310 genital tracts of cows were collected from slaughter house of Dinajpur Sadar Upazilla during April 2009 to March 2010. Among the 310 samples, 31.29% (n=97/310) were affected with endrometritis. Similarly, 8.37% (n=26/310) cystic ovary, 6.77% (n=21/310) ovary hyperplasia, 4.84% (n=15/310) pyometra, 4.84% (n=15/310) parovarian cyst, 4.52% (n=14/310) hydrometra, 4.84% (n=15/310) ovary hypoplasia, 3.55%(n=11/310) ovaro-bursal adhesion, 1.29% (n=4/310) vaginal cyst and 0.66% (n=2/310)) hemorrhagic uterine horn were detected by post-mortem examination, the cases were reconfirmed by histopathological studies. Uterine fluid (n=50) samples were collected, and were subjected for conventional bacteriological culture and biochemical analysis. Escherichia coli and Salmonella sp. could be identified from 30% (n=15/50) and 8% (n=4/50) samples, respectively. In conclusion, various pathological disorders in the female reproductive system of cows are prevalent, that may cause reduction of calf production.

Objective: Sertoli cells (SCs) are important sustentacular cells in the seminiferous tubules of the testis. Isolation and identification of SCs are the premise for studying their functions. Since New Zealand rabbit is a stable strain which is widely used for biomedical research and animal farming, this study aimed to develop a simple and effective protocol for SC isolation in New Zealand rabbits. Materials and Methods: The SCs of three 30-day-old New Zealand rabbits were isolated by incu¬bation with enzymatic digestion I (Dulbeccos modified Eagle medium supplemented with 1 mg/ ml collagenase IV and 50 μg/ml DNase I) and digestion II (digestion I + 1 mg/ml hyaluronidase + 1 mg/ml trypsin), as well as differential plating. The cells were enriched and identified by using immunocytochemical staining and reverse transcription polymerase chain reaction analysis. Results: Homogeneous cells were obtained. They presented the typical large cell body and an irregular pyramidal shape after differential plating and passaging. These cells expressed mRNA of the SC marker sex-determining region Y-box 9 (SOX9) instead of the Leydig cell marker StAR. Immunocytochemically, they are positive of SOX9, GATA binding protein 4, and androgen-binding protein. Conclusion: The SCs were enriched from the testicular tissues of prepubertal New Zealand rabbits by a simple and effective protocol, which provides a basis for further theoretical researches and practical applications. [J Adv Vet Anim Res 2021; 8(2.000): 218-223]

Duck plague (DP) is the most feared duck disease in the world. For isolation, identification, molecular detection and characterization of DP virus (DPV), a total of 94 samples were collected from commercial farms (n=6) and households (n=13) from Rajshahi (n=37), Netrokona (n=35) and Mymensingh (n=22) districts of Bangladesh. The samples were processed and inoculated into 11-13 days old embryonated duck eggs for virus propagation. Virus was identified using agar gel immunodiffusion test (AGIT) and passive hemagglutination (PHA) test, and was confirmed by polymerase chain reaction (PCR) targeting DNA polymerase and gC genes, followed by sequencing. Pathogenicity tests were performed using duck embryos, ducklings and ducks. Among the 94 samples, 17 isolates were confirmed as DPV by PCR amplification of partial DNA polymerase (446-bp) and gC genes (78-bp), respectively. One of the isolates (Anatid herpes 1 BAU DMH) was sequenced and found to be closely related with a Chinese variant of DPV (GenBank: JQ647509.1). Thus, we assume that both Bangladeshi and Chinese isolates of DPV may have a common ancestor. [J Adv Vet Anim Res 2015; 2(3.000): 296-303]