S.A. Wani

Isolation, molecular characterization and prevalence of Clostridium perfringens in sheep and goats of Kashmir Himalayas, India

Aim: The study was conducted to report the occurrence of the Clostridium perfringens in sheep and goats of the Kashmir valley for the 1st time and to characterize them molecularly with respect to toxin genes to determine the prevalence of the various toxinotypes. Materials and Methods: A total of 177 samples (152 from sheep and 25 from goats) collected from healthy, diarrheic animals, and morbid material of animals suspected to have died of enterotoxaemia were screened for C. perfringens toxinotypes. The presumptive positive isolates were confirmed using 16S rRNA gene-based polymerase chain reaction (PCR). All the confirmed isolates were screened for six toxin genes, namely; cpa, cpb, etx, cpi, cpb2, and cpe using a multiplex PCR. Results: The PCR amplification of 16S rRNA gene revealed that out of 177 samples collected, 125 (70.62%) were found positive for C. perfringens, of which 110 (72.36%) were from sheep and 15 (60%) were from goats. The highest prevalence of C. perfringens toxinotype D was observed in lambs (56.16%) and kids (46.16%) followed by 3.84% in adult sheep while it was absent in samples obtained from adult goats. The multiplex PCR revealed that 67 (60.90%) isolates from sheep and 8 (53.33%) isolates from goats belonged to toxinotype A, while 43 (39.09%) isolates from sheep and 7 (46.66%) isolates from goats were detected as toxinotype D. None of the isolates was found to be toxinotype B, C, or E. All the C. perfringens toxinotype A isolates from sheep were negative for both cpb2 and cpe genes, however, 27.90% toxinotype D isolates from sheep carried cpb2 gene, and 6.97% possessed cpe gene. In contrast, 12.50% C. perfringens toxinotype A isolates from goats harbored cpb2 and cpe genes while 14.28% isolates belonging to toxinotype D carried cpb2 and cpe genes, respectively. Conclusion: The high prevalence of C. perfringens was observed, even in day-old lambs. The toxinotypes A and D are prevalent in both sheep and goats. The severity of disease and mortality may be associated with the presence of minor toxins in both the detected toxinotypes.

Molecular Detection and Isolation of Mycoplasma capricolum subsp. capripneumoniae in Pashmina and Local Goats in Five Districts of Kashmir, India -

Contagious caprine pleuropneumonia (CCPP) is caused by Mycoplasma capricolum subsp. caprip-neumoniae (Mccp), which belongs to Mycoplasma mycoides cluster, a group of closely related Mycoplasmas that are pathogenic to ruminants. The clinical lesions of CCPP are restricted to alveolar tissues of infected goats, which distinguish it from other respiratory diseases of small ruminants caused by members of the Mycoplasma mycoides cluster. The outbreaks of CCPP in Pashmina goats have been reported in cold arid regions of Kashmir province, such as Leh and Kargil districts and northern parts of the Kashmir valley. In the current study, a total of 55 lung samples and 70 nasal swabs were collected from clinically sick goats showing respiratory symptoms prior to death and processed for Mccp isolation and identification. The Mccp was detected in 33 (60.0%) and 24 (34.2%) samples, respectively by PCR. All the PCR positive samples for Mccp were further processed for isolation but only five isolates were obtained due to their fastidious nature which were confirmed by typical colony morphology and DNA sequencing of 548 bp 16SrRNA gene fragment. The five isolates were isolated from lung samples of Pashmina goats of Leh district. The results of the current study reveal that Pashmina goats of Ladakh region are highly vulnerable to CCPP and is the major cause of mortality and morbidity among them particularly during the winter season. Hence, there is an immediate need to isolate and characterise the organism for the development of autogenous vaccine to combat this economically important disease of goats. This communication records the first ever report on molecular detection and isolation of Mccp from Pashmina goats in India and is the first step towards development of vaccine against CCPP.

Clostridium perfringens Type A from Broiler Chicken with Necrotic Enteritis in Kashmir Valley, India

Avian necrotic enteritis (NE) caused by C. perfringens type A, rod shaped, grampositive, anaerobic spore forming bacterium is wide spread in broilers imposing a significant economic burden on the poultry industry worldwide (Dahiya et al., 2006). It is an acute, often fatal, disease of chickens characterized by depression, loss of appetite and sudden death. It occurs both as an acute clinical disease and as a subclinical disease with necrosis in the intestines or as C. perfringens-associated hepatitic change (CPH) witiohepatitis or fibrinoid necrosis in the liver. In broilers, outbreaks of clinical NE have been reported from all poultry growing areas of the world (Ficken and Wages, 1997; Dahiya et al., 2006). It has also been recorded in wild geese (Wobeser and Rainnie, 1987), wild crows (Asaoka et al., 2004), ostriches (Kwon et al., 2004) and in capercaillies reared in captivity (Hofshagen and Stenwig, 1992). C. perfringens type A from soil, dust and contaminated feed and litter has been implicated as a source of infection (Craven et International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 12 (2017) pp. 2443-2453 Journal homepage: http://www.ijcmas.com