Amplification and cloning of exon regions from 1-4 of cathepsin B-like cysteine protease gene of Haemonchus contortus and immune response in lambs against cathepsin B-like cysteine protease fraction

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  • CHARU PATHAK Senior Research Fellow, ICAR-Directorate of Knowledge Management in Agriculture, KAB-I, Pusa, New Delhi
  • S K GHOURI Principal Scientist, Maharaja Ganga Singh University of Bikaner, Bikaner, Rajasthan 334 001 India
  • S K KUMAR Scientist, National Research Center on Camel, Jorbeer
  • M M SAXENA Dean, Faculty of Science and Director Research, Maharaja Ganga Singh University, Bikaner


Cysteine proteases, Haemonchus contortus, Small ruminants, Vaccine


Haemonchus contortus is an economically important, highly pathogenic blood-sucking nematode parasite of ruminants. A study was conducted to characterize the cathepsin B-like cysteine protease gene of H. contortus of Indian origin, which has been explored as potential target for vaccine development. Adult H. contortus worms were collected from the abomasum of sheep and goats and whole genomic DNA was extracted and amplified using appropriate primers. The expected amplicon of 767 bp was cloned into T-vector using GeNeiTM Instant cloning kit. Immunization study was conducted to examine the effect of cysteine protease fraction on the faecal egg counts in lambs. It was found that the egg per gram of faeces was lower in immunized group as compared to control group (at 10, 11, 12, and 15 week). At post mortem, the worms recovered from the abomasa were lower in immunized group (800.40±101.8) as compared to control group (950.93±300.29). The results indicated that the immunization with cysteine protease fraction in lambs reduces the faecal egg count, but did not remove all the worms from lambs.


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Burke J M and Miller J E. 2006. Evaluation of multiple low doses of copper oxide wire particles compared with levamisole for control of Haemonchus contortus in lambs. Veterinary Parasitology 139: 145–49. DOI:

Cheng M, Yang X, Li Z, He H, Qu Z, He A, Wu Z, and Zhan X. 2012. Cloning and characterization of a novel cathepsin Blike cysteine proteinase from Angiostrongylus cantonensis. Parasitology Research 110: 2413–22. DOI:

Dalton J P, Brindley P J, Knox D P, Brady C P, Hotez P J, Donnelly S, O’neill SM, Mulcahy G and Loukas A. 2003. Helminth vaccines: from mining genomic information for vaccine targets to systems used for protein expression. International Journal of Parasitology 33: 621–40. DOI:

Das M and Singh S. 2005. Anthelminthic resistance to nematodes in sheep and goat farms in Hissar. Journal of Veterinary Parasitology 19: 103–06.

De Vries E, Bakker N, Krijgsveld J, Knox D P, Heck A J and Yatsuda A P. 2009. An AC-5 cathepsin B-like protease purified from Haemonchus contortus excretory secretary products shows protective antigen potential for lambs. Veterinary Research 40: 41. DOI:

Dobson R J, Waller P J and Donald A D. 1990. Population dynamics of Trichostrongylus columbiformis in sheep: the effect of infection rate on the establishment of infective larvae and parasite fecundity. International Journal of Parasitology 46: 205–13.

Donald A D, Morley F H W, Waller PJ, Axelsen A and Donnelley J R. 1978. Availability of grazing sheep of gastrointestinal nematode infection arising from summer contamination of pastures. Australian Journal of Agricultural Research 29: 189– 204. DOI:

Gupta R P, Yadav C L and Chaudhry S S. 1987. Epidemiology of gastrointestinal nematodes of sheep and goats in Haryana, India. Veterinary Parasitology 24: 117–27. DOI:

Jacquiet P, Humbert J F, Comes A M, Cabaret J, Thiam A and Cheikh D. 1995. Ecological, Morphological and Genetic characterization of sympatric Haemonchus spp. Parasites of domestic ruminants in Mauritania. Parasitology 110: 483– 92. DOI:

Jasmer D P, Mitreva M D and McCarter J P. 2004. mRNA sequences for Haemonchus contortus intestinal cathepsin Blike cysteine proteases display an extreme in abundance and diversity compared with other adult mammalian parasitic nematodes. Molecular and Biochemical Parasitology 137: 297–305. DOI:

Jasmer D P, Roth J and Myler P J. 2001. Cathepsin B-like cysteine proteases and Caenorhabditis elegans homologues dominate gene products expressed in adult Haemonchus contortus intestine. Molecular and Biochemical Parasitology 116: 159– 69. DOI:

Jithendran K P and Bhat T K. 1999. Epidemiology of parasitoses in dairy animals in North-West humid Himalayan region of India with particular reference to gastrointestinal nematodes. Tropical Animal Health and Production 31: 205–14. DOI:

Knox D P, Redmond D L, Newlands G F, Skuce P J, Pettit D and Smith WD. 2003. The nature and prospects for gut membrane proteins as vaccine candidates for Haemonchus contortus and other ruminant trichostrongyloids. International Journal of Parasitology 33: 1129–37. DOI:

Lee J, Kim J H, Sohn H J, Yang H J, Na B K, Chwae Y J, Park S, Kim K, and Shin H J. 2014. Novel cathepsin B and cathepsin B-like cysteine protease of Naegleria fowleri excretorysecretory proteins and their biochemical properties. Parasitology Research 113: 2765–76. DOI:

Longbottom D, Redmond D L, Russel M, Liddell S, Smith W D and Knox D P. 1997. Molecular cloning and characterization of a putative aspartate proteinase associated with a gut membrane protein complex from adult Haemonchus contortus. Molecular and Biochemical Parasitology 88: 63–72. DOI:

MAFF. 1986. Manual of veterinary parasitological laboratory techniques. Part 1 Helminthology. Reference Book No. 418, Ministry of Agriculture, Fisheries and Food.

Pathak C, Ghouri S K, Kumar S and Saxena M M. 2012. Haemonchus contortus isolate Bikaner cysteine proteinase mRNA. NCBI GeneBank Accession No. JX402408.1

Pratt D, Cox G N, Milhausen M J and Boisvenue R J. 1990. A developmentally regulated cysteine protease gene family in Haemonchus contortus. Molecular and Biochemical Parasitology 43: 181–91. DOI:

Redmond D L, Knox D P, Newlands G and Smith W D. 1997. Molecular cloning and characterization of a developmentally regulated putative metallopeptidase present in a host protective extract of Haemonchus contortus. Molecular and Biochemical Parasitology 85: 77–87. DOI:

Sajid M and Mckerrow J H. 2002. Cysteine proteases of parasitic organisms. Molecular and Biochemical Parasitology 120: 1–21. DOI:

Sambrook J and Russel D W. 2001. Molecular cloning: A Laboratory Manual. 3rd edn, Cold Spring Harbor Laboratory Press, New York.

Sanyal P K. 1998. Integrated parasite management in ruminants in India. Biological control of gastrointestinal nematodes in ruminants using predacious fungi (FAO). FAO Animal Production and Health Paper 144: 54–65.

Singh D, Swarnkar CP and Khan F A. 2002. Anthelminthic resistance in gastrointestinal nematodes of livestock in India. Journal of Veterinary Parasitology 16: 115–30.

Singla L D. 1995. A note on sub-clinical gastro-intestinal parasitism in sheep and goats in Ludhiana and Faridkot districts of Punjab. Indian Veterinary Medical Journal 19: 61–62.

Smith W D, Smith S K, Pettit D, Newlands G F and Skuce P J. 2000. Relative protective properties of three membrane glycoprotein fraction from Haemonchus contortus. Parasite Immunology 22: 63–71. DOI:









How to Cite

PATHAK, C., GHOURI, S. K., KUMAR, S. K., & SAXENA, M. M. (2015). Amplification and cloning of exon regions from 1-4 of cathepsin B-like cysteine protease gene of Haemonchus contortus and immune response in lambs against cathepsin B-like cysteine protease fraction. The Indian Journal of Animal Sciences, 85(11), 1167–1171.