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Nazki, Salik
- Isolation, Molecular Characterization and Prevalence of Clostridium perfringens in Sheep and Goats of Kashmir Himalayas, India
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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.
Authors
Salik Nazki
1,
Shakil A. Wani
2,
Rafia Parveen
2,
Showkat A. Ahangar
2,
Zahid A. Kashoo
2,
Syed Hamid
3,
Zahoor A. Dar
2,
Tanveer A. Dar
4,
Pervaiz A. Dar
2
Affiliations
1 Laboratory of Veterinary Immunology, College of Veterinary Medicine, Chonbuk National University, Iksan-54596, KR
2 Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, SKUAST-Kashmir, Shuhama, Srinagar-190006, Jammu and Kashmir, IN
3 Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, SKUAST-Jammu, R.S. Pora, Jammu-181102, IN
4 Division of Veterinary Pathology, SKUAST-Kashmir, Shuhama, Srinagar-190006, Jammu and Kashmir, IN
1 Laboratory of Veterinary Immunology, College of Veterinary Medicine, Chonbuk National University, Iksan-54596, KR
2 Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, SKUAST-Kashmir, Shuhama, Srinagar-190006, Jammu and Kashmir, IN
3 Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, SKUAST-Jammu, R.S. Pora, Jammu-181102, IN
4 Division of Veterinary Pathology, SKUAST-Kashmir, Shuhama, Srinagar-190006, Jammu and Kashmir, IN
Source
Veterinary World, Vol 10, No 12 (2017), Pagination: 1501-1507Abstract
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.
Keywords
Clostridium perfringens, Enterotoxemia, Multiplex Polymerase Chain Reaction, Toxinotype, 16S rRNA.Full Text
- Phenotypic and Genotypic Characterization of Methicillin-Resistant Staphylococcus aureus from Bovine Mastitis
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Materials and Methods: A total of 160 mastitic milk samples were screened for the presence of S. aureus. The presumptive positive isolates were confirmed using nuc and 23S rRNA gene-based polymerase chain reaction. All the confirmed isolates were subjected to in vitro antibiogram using a number of antibiotics. Isolates which showed resistance against methicillin were characterized for the presence of mecA gene.
Results: Out of the total 160 milk samples, 36 (22.5%) samples yielded S. aureus. The in vitro antibiogram revealed that 16.6% S. aureus isolates were resistant to all antibiotics screened for and 5.5% isolates were sensitive to all of them. Furthermore, the study found 94.4%, 83.3%, 77.7%, 66.6%, 50%, and 27.7% of S. aureus isolates resistant to penicillin, ampicillin, amoxicillin-sulbactam, enrofloxacin, ceftriaxone, and methicillin, respectively. Out of the 36 S. aureus isolates, only 6 (16.6%) isolates were confirmed as MRSA while rest were MSSA.
Conclusion: The higher occurrence of S. aureus-mediated mastitis was concluded due to improper hygienic and poor farm management. The multiple drug resistance reveals the indiscriminate use of drugs and presence of methicillin resistance gene determinant is an alarming situation as such infections are difficult to treat.
Authors
S. Hamid
1,
Mohd Altaf Bhat
1,
Irfan Ahmad Mir
1,
Anil Taku
1,
Gulzar Ahmad Badroo
1,
Salik Nazki
2,
Andleeb Malik
1
Affiliations
1 Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Jammu and Kashmir, IN
2 Laboratory of Veterinary Immunology, College of Veterinary Medicine, Chonbuk National University, Iksan, KR
1 Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Jammu and Kashmir, IN
2 Laboratory of Veterinary Immunology, College of Veterinary Medicine, Chonbuk National University, Iksan, KR
Source
Veterinary World, Vol 10, No 3 (2017), Pagination: 363-367Abstract
Aim: This study was conducted to determine the occurrence of methicillin-sensitive and Staphylococcus aureus (MSSA) methicillin-resistant S. aureus (MRSA) from bovine mastitis and to characterize them with respect to antibiotic resistance gene mecA.Materials and Methods: A total of 160 mastitic milk samples were screened for the presence of S. aureus. The presumptive positive isolates were confirmed using nuc and 23S rRNA gene-based polymerase chain reaction. All the confirmed isolates were subjected to in vitro antibiogram using a number of antibiotics. Isolates which showed resistance against methicillin were characterized for the presence of mecA gene.
Results: Out of the total 160 milk samples, 36 (22.5%) samples yielded S. aureus. The in vitro antibiogram revealed that 16.6% S. aureus isolates were resistant to all antibiotics screened for and 5.5% isolates were sensitive to all of them. Furthermore, the study found 94.4%, 83.3%, 77.7%, 66.6%, 50%, and 27.7% of S. aureus isolates resistant to penicillin, ampicillin, amoxicillin-sulbactam, enrofloxacin, ceftriaxone, and methicillin, respectively. Out of the 36 S. aureus isolates, only 6 (16.6%) isolates were confirmed as MRSA while rest were MSSA.
Conclusion: The higher occurrence of S. aureus-mediated mastitis was concluded due to improper hygienic and poor farm management. The multiple drug resistance reveals the indiscriminate use of drugs and presence of methicillin resistance gene determinant is an alarming situation as such infections are difficult to treat.
Keywords
Bovine, Mastitis, Methicillin-Resistant Staphylococcus aureus, mecA, Methicillin-Sensitive Staphylococcus aureus, Staphylococcus aureus.Full Text