12-Title: Molecular identification of epsilon gene and pathomorphology of intestine in sheep with reference to Clostridium perfringens type D

12-Title: Molecular identification of epsilon gene and pathomorphology of intestine in sheep with reference to Clostridium perfringens type D

Authors: Renu, H Dadhich, M Mathur, PK Boyal and Subhita Kumari

Source: Ruminant Science (2022)-11(1):53-56.

How to cite this manuscript: Renu, Dadhich H, Mathur M, Boyal PK and Kumari Subhita (2022). Molecular identification of epsilon gene and pathomorphology of intestine in sheep with reference to Clostridium perfringens type D. Ruminant Science 11(1):53-56.


The aim of this study was to determine molecular identification of epsilon toxin secreted by Clostridium perfringens type D from intestine of sheep that died due to enterotoxemia and pathological changes observed in intestine.  Necropsy was performed on 362 sheep irrespective of age, sex and breeds. Among these, 123 sheep suspected for enterotoxemia were subjected detection of Etx toxin gene of Clostridium perfringens type D from intestinal contents by using PCR. PCR results of 66 sheep samples found positive for Clostridium perfringens type D.  PCR positive sheep intestinal tissues were further processed for histopathological studies. Grossly, affected intestines were congested, hemorrhagic and filled with gas. Microscopically, intestine showed goblet cells hyperplasia, edema, haemorrhages, blunted and fused villi, mononuclear cells and polymorphonuclear cells infiltration. In conclusion, Clostridium perfringens type D associated enterotoxemia is a fatal enteric disease which can be identified by histopathological alteration and molecular identification.


Ahsani MR, Mohammadabadi MR, Shamsaddini MB (2010). Clostridium perfringens isolate typing by multiplex PCR. Journal of Venomous Animals and Toxins 16(4):573-578.

Bullen JJ and Battey I (1957). Enterotoxaemia of sheep. Veterinary Record 69:1268-1273.

Elsify A, Tarabess R, Nayel MA, Salama A, Allaam M, El-Gaber AM, Hassan H, Zaghawa A and Elballal S (2016). Bacteriological and molecular studies on Clostridium perfringens isolated from sheep in three Egyptian provinces. African Journal of Microbiology Research 10(20):725-732.

Fernandez Miyakawa ME and Uzal FA (2003). The early effects of Clostridium perfringens type D epsilon toxin in ligated intestinal loops of goats and sheep. Veterinary Research Communications 27:231-241.

Hart ML, Meyer A, Johnson PJ and Ericsson AC (2015). Comparative evaluation of DNA extraction methods from feces of multiple host species for downstream next-generation sequencing. PLoS One 10(11):1-16.

Hassanein KMA, Sayed MM and Hassan AM (2017). Pathological and biochemical studies on enterotoxemia in sheep. Comparative Clinical Pathology 26:513-518.

Hussain K, Ijaz M, Farooqi SH, Rizvi SNB, Ali A, Ghaffar A (2018). Molecular Characterization of Clostridium perfringens Toxino-types and Type ‘D’ Multidrug Resistance Profile in Diarrheic Sheep. Pakistan Veterinary Journal 38(3):271-275.

Jubb, Kennedy and Palmer’s (2007). Pathology of Domestic Animals. 5th Edn, Elsevier Health Sciences. pp 218-1220.

Kumar R, Gururaj K, Pawaiya RVS, Mishra AK, Chaturvedi V, Varshney M, Andani D, Jena D, Sharma A, Gangwar NK and Singh R (2019). Pathology of experimental Clostridium perfringens type D enterotoxaemia in goats. Indian Journal of Veterinary Pathology 43(2):94-103.

Kumar VN, Sreenivasulu D and Reddy YN (2014). Prevalence of Clostridium perfringens toxin genotypes in enterotoxemia suspected sheep flocks of Andhra Pradesh. Veterinary World 7(12):1132-36.

Lillie RD (1965). Histopathological Technique and Practical Histochemistry. McGraw Hill Book Co; New York and London.

Mekathoti RD (2018). Pathological and molecular diagnosis of Clostridium perfringens type D infection in sheep. MVSc thesis submitted to Sri Venkateswara Veterinary University, Tirupati.

Nazki S, Wani SA, Parveen R, Ahangar SA, Kashoo ZA, Hamid S, Dar ZA, Dar TA and Dar PA (2017). Isolation, molecular characterization and prevalence of Clostridium perfringens in sheep and goats of Kashmir Himalayas, India. Veterinary World 10(12):1501-1507.

Popoff MR and Bouvet P (2009). Clostridial toxins. Future Microbiology 4(8):1021-64.

Rasool S, Hussain I, Wani SA, Kashoo ZA, Beigh Q and Nyrah Q (2017). Molecular Typing of Clostridium perfringens Isolates from Faecal Samples of Healthy and Diarrhoeic Sheep and Goats in Kashmir. International Journal of Current Microbiology and Applied Sciences. 2017;6(10):3174-3184.

Salvarani FM, Faccin M, Freitas NFdeQR, Matos MRde, Garcia EC, Pagliosa GM, Barbosa JD, Lobato FCF and Viott AdeM (2019). Outbreak of Clostridium perfringens type D enterotoxemia in sheep. Veterinary Medicine 40(6):2593-2602.

Singh DD, Pawaiya RVS, Gururaj K, Gangwar NK, Mishra AK, Singh R, Pathak A, Andani D, Gautam TK and Kumar A (2017). Pathological studies on spontaneous cases of enterotoxaemia in goat kids at organized and unorganized farms. Indian Journal of Veterinary Pathology 41(4):245-250.