Infection Dynamics of Clostridium perfringens Fingerprinting in Buffalo and Cattle of Punjab Province, Pakistan

Muhammad Umar Zafar Khan; Shumaila Khalid; Muhammad Humza; Shunli Yang; Mughees Aizaz Alvi; Tahir Munir; Waqar Ahmad; Muhammad Zahid Iqbal; Muhammad Farooq Tahir; Yongsheng Liu; Jie Zhang

doi:10.3389/fvets.2022.762449

Infection Dynamics of Clostridium perfringens Fingerprinting in Buffalo and Cattle of Punjab Province, Pakistan

Front Vet Sci. 2022 Jul 22:9:762449. doi: 10.3389/fvets.2022.762449. eCollection 2022.

Authors

Muhammad Umar Zafar Khan^{1

2}, Shumaila Khalid³, Muhammad Humza^{4

5}, Shunli Yang^{6

7}, Mughees Aizaz Alvi⁸, Tahir Munir³, Waqar Ahmad⁹, Muhammad Zahid Iqbal¹⁰, Muhammad Farooq Tahir¹¹, Yongsheng Liu¹, Jie Zhang¹

Affiliations

¹ Hebei Key Laboratory of Preventive Veterinary Medicine, College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao, China.
² Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan.
³ Livestock and Dairy Development Department, Lahore, Pakistan.
⁴ Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing, China.
⁵ Department of Plant Pathology, University of Agriculture, Faisalabad, Pakistan.
⁶ State Key Laboratory of Veterinary Etiological Biology of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
⁷ Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, China.
⁸ Department of Clinical Medicine and Surgery, University of Agriculture, Faisalabad, Pakistan.
⁹ The Equine Clinic, Al-Hashar Stables, Muscat, Oman.
¹⁰ Department of Veterinary Medicine, University of Veterinary and Animal Sciences, Lahore, Pakistan.
¹¹ Health Security Partners, Washington, DC, United States.

Abstract

Clostridium perfringens produces core virulence factors that are responsible for causing hemorrhagic abomasitis and enterotoxemia making food, animals, and humans susceptible to its infection. In this study, C. perfringens was isolated from necropsied intestinal content of buffalo and cattle belonging to four major bovine-producing regions in the Punjab Province of Pakistan for the purpose offind out the genetic variation. Out of total 160 bovine samples (n: 160), thirty-three (n: 33) isolates of C. perfringens were obtained from buffalo (Bubales bubalis) and cattle (Bos indicus) that were further subjected to biochemical tests; 16S rRNA based identification and toxinotyping was done using PCR (Polymerase Chain Reaction) and PFGE (Pulse Field Gel Electrophoresis) pulsotypesfor genetic diversity. Occurrence of C. perfringens was found to be maximum in zone-IV (Bhakkar and Dera Ghazi Khan) according to the heatmap. Correlation was found to be significant and positive among the toxinotypes (α-toxin, and ε-toxin). Response surface methodology (RSM) via central composite design (CCD) and Box-Behnken design (BBD) demonstrated substantial frequency of C. perfringens based toxinotypes in all sampling zones. PFGE distinguished all isolates into 26 different pulsotypes using SmaI subtyping. Co-clustering analysis based on PFGE further decoded a diversegenetic relationship among the collected isolates. This study could help us to advance toward disease array of C. perfringens and its probable transmission and control. This study demonstrates PFGE patterns from Pakistan, and typing of C. perfringens by PFGE helps illustrate and mitigate the incidence of running pulsotypes.

Keywords: Clostridium perfringens; PFGE; intestinal contents; sampling zones; toxinotyping.