Molecular Detection and Characterization of Haemonchus spp. in Cattle in Nigeria
Abstract
Background: Haemonchosis is a major parasitic infestation in ruminant livestock, causing significant economic losses annually. The causative organisms are helminths of the genus Haemonchus spp. Detection of the causative agent is important for effective management and control of the disease. Molecular detection and characterization of parasites is a very dependable approach for parasite identification, especially where morphological characterization is unreliable.
Methods: To detect and characterize Haemonchus species in cases of haemonchosis at a Municipal abattoir in Ibadan, Nigeria; abomasal samples were collected from cattle at the abattoir. Polymerase chain reaction (PCR) was used to detect and amplify 320 bp internal transcribed spacer-2 (ITS-2) and 400 bp external transcribed spacer (ETS) genes of the adult worms in the samples. Multiple sequence alignment and phylogenetic tree reconstruction were carried out to further confirm the presence of the worms.
Results: PCR, multiple sequence alignment, and phylogenetic reconstruction confirmed the presence of H. placei in the abomasal samples and further confirmed the species as a distinct species of bovine worms at the abattoir. Multiple sequence alignment also revealed genetic sites that can be employed to distinguish H. placei from H. contortus and H. similis.
Conclusion: Molecular techniques; PCR and sequence analysis are very important and reliable in the diagnosis of parasitic diseases. This will help to formulate effective control measures for eradication of the parasite.
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2. Rudolphi CA. New Observations on Intestinal Worms. Arch Zool Comp Anat. 1803; 3: 1-32.
3. Place FE. Anaemic diarrhoea in young cattle. Vet Rec. 1893; 5: 589.
4. Travassos LP. Trichostrongylideos brazileiros (III preliminary note). Brazil-Med. 1914; 28(34): 325-327.
5. Achi YL, Zinsstag J, Yao K, Yeo N, Dorchies P, Jacquiet P. Host specificity of Haemonchus spp. for domestic ruminants in the savanna in northern Ivory Coast. Vet Parasitol. 2003; 116(2): 151-158.
6. Lichtenfels JR, Pilitt PA, Hoberg EP. New morphological characters for identifying individual specimens of Haemonchus spp. (Nematoda: Trichostrongyloidea) and a key to species in ruminants of North America. J Parasitol. 1994; 80(1): 107-119.
7. Alessandro F, Amarante T. Why is it important to correctly identify Haemonchus species? (Colégio Brasileiro de Parasitologia Veterinaria). Rev Bras Parasitol Vet. 2011; 20(4): 263-268.
8. Lichtenfels JR, Pilitt PA, Le Jambre LF. Spicule lengths of the ruminant stomach nematodes Haemonchus contortus, Haemonchus placei, and their hybrids. Proc Helminthol Soc Wash. 1988; 55: 97-100.
9. Ali Q, Rashid MI, Ashraf K, Zahid MN, Ashraf S, Chaudhry U. Genetic variation in the rDNA ITS-2 sequence of Haemonchus placei from cattle host. J Inf Mol Biol. 2015; 3(1): 13-18.
10. Jeremiah OT, Banwo OG. Haematologic profile and prevalence survey of haemonchosis in various breeds of slaughtered cattle in Ibadan, Nigeria. Nig J Anim Prod. 2019; 46(3): 151-162.
11. Odeniran PO, Jegede HO, Adewoga TOS. Prevalence and risk perception of adult-stage parasites in slaughtered food animals (cattle, sheep and goat) among local meat personnel in Ipata abattoir, Ilorin, Nigeria. Vet Med Anim Sci. 2016; 4:1.
12. Almeida FA. Multiple resistance to anthelmintics by Haemonchus contortus and Trichostrongylus colubriformis in sheep in Brazil. Parasitol Int. 2010; 59(4): 622-625.
13. Sutherland IA, Leathwick DM. Anthelmintic resistance in nematode parasites of cattle: a global issue? Trends Parasitol.2011;27(4):176-181.
14. Hoberg EP, Lichtenfels JR, Gibbons L. Phylogeny for species of the genus Haemonchus (Nematoda: Trichostrongylo-idea): considerations of their evolutionary history and global biogeography among Camelidae and Pecora (Artiodactyla). J Parasitol. 2004; 90(5): 1085-1102.
15. Almeida JL. Note on the species of the genus Haemonchus Cobb, 1898 (Nematoda: Trichostrongyloidea). Proceedings of the Society of Biology and Its Branches. 1935; 114: 960–961.
16. Gibbons LM. Revision of the genus Haemonchus Cobb, 1898 (Nematoda: Trichostronglidae). Syst Parasitol. 1979; 1: 3-24.
17. Amarante AF, Bagnola J, Amarante MR, Barbosa MA. Host specificity of sheep and cattle nematodes in Sao Paulo state, Brazil. Vet Parasitol. 1997; 73(1-2): 89–104.
18. Jacquiet P, Cabaret J, Thiam E, Cheikh D. Host range and the maintenance of Haemonchus spp. in an adverse arid climate. Int J Parasitol. 1998; 28(2): 253–261.
19. Le Jambre LF. Hybridization studies of Haemonchus contortus (Rudolphi. 1803) and H. placei (Place, 1893) (Nematoda: Trichostrongylidae). Int J Parasitol. 1979;9: 455-463.
20. Stevenson LA, Chilton NB, Gasser RB. Differentiation of Haemonchus placei from H. contortus (Nematoda: Trichostrongylidae) by the ribosomal DNA second internal transcribed spacer. Int J Parasitol. 1995; 25(4): 483-488.
21. Lasisi OT, Ojo NA, Otesile EB. Estimation of age of cattle in Nigeria using rostral dentition. Trop Vet. 2002; 20: 204-208.
22. Hansen J, Perry B. The Epidemiology, Diagnosis and Control of Helminthes Parasites of Ruminants. A Handbook, ILRAD. 1994.
23. Cerutti MC, Citterio CV, Bazzocchi C. et al. Genetic variability of Haemonchus contortus (Nematoda: Trichostrongyloidea) in alpine ruminant host species. J Helminthol. 2010; 84(3): 276–283.
24. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic Local Alignment Search Tool. J Mol Biol. 1990; 215(3): 403–410.
25. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximumlikelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011; 28(10): 2731–2739.
26. Palevich N, Britton C, Kamenetzky L, Mitreva M, de Moraes MM, Bennuru S, et al. Tackling hypotheticals in helminth genomes. Trends Parasitol. 2018; 34: 179–83.
27. Ndosi BA, Lee D, Bia MM, et al. Morphometry and Molecular Identification of Haemonchus Cobb, 1898 (Trichostrongylidae: Nematoda) Isolates from Small Ruminants in Tanzania Based on Mitochondrial cox 1 and rRNA-ITS genes. J Parasitol Res. 2023; 2023:1923804.
28. Mochizuki R, Endoh D, Onuma M, Fukumoto S. PCR-based species-specific amplification of ITS of Mecistocirrus digitatus and its application in identification of GI nematode eggs in bovine faeces. J Vet Med Sci. 2006; 68(4): 345-351.
29. Food and Agricultural Organization (FAO). “Distribution and impact of helminth diseases of livestock in developing countries,” in FAO Corporate Document Repository— Agriculture and Consumer Protection. 2000.
30. Zarlenga DS, Chute MB, Martin A, Kapel CMO. A multiplex PCR for unequivocal differentiation of all encapsulated and non-encapsulated genotypes of Trichinella. Int J Parasitol. 1999; 29(11): 1859-1867.
31. Ademola IO, Krücken J, Ramünke S, Demeler J, Von G, Himmelstjerna S. Absence of detectable benzimidazole resistance associated alleles in Haemonchus placei in cattle in Nigeria revealed by pyrosequencing of β-tubulin isotype 1. Parasitol Res. 2015;114(5):1997-2001.
32. Rocha RA, Rocha R, Bresciani Ket al. Sheep and cattle grazing alternately: Nematode parasitism and pasture decontamination. Small Rumin Res. 2008;75(2-3):135-143.
33. Nascimento AA, Bonuti MR, Mapeli EB, et al. Natural infections in cervids (Mammalia: Cervidae) from the states of Mato Grosso do Sul and São Paulo by Trichostrongyloidea nematodes. Braz J Vet Res Anim Sci. 1927;37(2):153-158.
34. Zarlenga DS, Stringfellow F, Nobary M, Lichtenfels JR. Cloning and characterization of ribosomal RNA genes from three species of Haemonchus (Nematoda: Tric-hostrongyloidea) and identification of PCR primers for rapid differentiation. Exp Parasitol. 1994; 78(1): 28-36.
35. Palevich N, Maclean PH, Choi YJ, Mitreva M. Characterization of the complete mitochondrial genomes of two sibling species of parasitic roundworms, Haemonchus contortus and Teladorsagia circumcincta. Front Genet. 2020;11: 573395.
2. Rudolphi CA. New Observations on Intestinal Worms. Arch Zool Comp Anat. 1803; 3: 1-32.
3. Place FE. Anaemic diarrhoea in young cattle. Vet Rec. 1893; 5: 589.
4. Travassos LP. Trichostrongylideos brazileiros (III preliminary note). Brazil-Med. 1914; 28(34): 325-327.
5. Achi YL, Zinsstag J, Yao K, Yeo N, Dorchies P, Jacquiet P. Host specificity of Haemonchus spp. for domestic ruminants in the savanna in northern Ivory Coast. Vet Parasitol. 2003; 116(2): 151-158.
6. Lichtenfels JR, Pilitt PA, Hoberg EP. New morphological characters for identifying individual specimens of Haemonchus spp. (Nematoda: Trichostrongyloidea) and a key to species in ruminants of North America. J Parasitol. 1994; 80(1): 107-119.
7. Alessandro F, Amarante T. Why is it important to correctly identify Haemonchus species? (Colégio Brasileiro de Parasitologia Veterinaria). Rev Bras Parasitol Vet. 2011; 20(4): 263-268.
8. Lichtenfels JR, Pilitt PA, Le Jambre LF. Spicule lengths of the ruminant stomach nematodes Haemonchus contortus, Haemonchus placei, and their hybrids. Proc Helminthol Soc Wash. 1988; 55: 97-100.
9. Ali Q, Rashid MI, Ashraf K, Zahid MN, Ashraf S, Chaudhry U. Genetic variation in the rDNA ITS-2 sequence of Haemonchus placei from cattle host. J Inf Mol Biol. 2015; 3(1): 13-18.
10. Jeremiah OT, Banwo OG. Haematologic profile and prevalence survey of haemonchosis in various breeds of slaughtered cattle in Ibadan, Nigeria. Nig J Anim Prod. 2019; 46(3): 151-162.
11. Odeniran PO, Jegede HO, Adewoga TOS. Prevalence and risk perception of adult-stage parasites in slaughtered food animals (cattle, sheep and goat) among local meat personnel in Ipata abattoir, Ilorin, Nigeria. Vet Med Anim Sci. 2016; 4:1.
12. Almeida FA. Multiple resistance to anthelmintics by Haemonchus contortus and Trichostrongylus colubriformis in sheep in Brazil. Parasitol Int. 2010; 59(4): 622-625.
13. Sutherland IA, Leathwick DM. Anthelmintic resistance in nematode parasites of cattle: a global issue? Trends Parasitol.2011;27(4):176-181.
14. Hoberg EP, Lichtenfels JR, Gibbons L. Phylogeny for species of the genus Haemonchus (Nematoda: Trichostrongylo-idea): considerations of their evolutionary history and global biogeography among Camelidae and Pecora (Artiodactyla). J Parasitol. 2004; 90(5): 1085-1102.
15. Almeida JL. Note on the species of the genus Haemonchus Cobb, 1898 (Nematoda: Trichostrongyloidea). Proceedings of the Society of Biology and Its Branches. 1935; 114: 960–961.
16. Gibbons LM. Revision of the genus Haemonchus Cobb, 1898 (Nematoda: Trichostronglidae). Syst Parasitol. 1979; 1: 3-24.
17. Amarante AF, Bagnola J, Amarante MR, Barbosa MA. Host specificity of sheep and cattle nematodes in Sao Paulo state, Brazil. Vet Parasitol. 1997; 73(1-2): 89–104.
18. Jacquiet P, Cabaret J, Thiam E, Cheikh D. Host range and the maintenance of Haemonchus spp. in an adverse arid climate. Int J Parasitol. 1998; 28(2): 253–261.
19. Le Jambre LF. Hybridization studies of Haemonchus contortus (Rudolphi. 1803) and H. placei (Place, 1893) (Nematoda: Trichostrongylidae). Int J Parasitol. 1979;9: 455-463.
20. Stevenson LA, Chilton NB, Gasser RB. Differentiation of Haemonchus placei from H. contortus (Nematoda: Trichostrongylidae) by the ribosomal DNA second internal transcribed spacer. Int J Parasitol. 1995; 25(4): 483-488.
21. Lasisi OT, Ojo NA, Otesile EB. Estimation of age of cattle in Nigeria using rostral dentition. Trop Vet. 2002; 20: 204-208.
22. Hansen J, Perry B. The Epidemiology, Diagnosis and Control of Helminthes Parasites of Ruminants. A Handbook, ILRAD. 1994.
23. Cerutti MC, Citterio CV, Bazzocchi C. et al. Genetic variability of Haemonchus contortus (Nematoda: Trichostrongyloidea) in alpine ruminant host species. J Helminthol. 2010; 84(3): 276–283.
24. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic Local Alignment Search Tool. J Mol Biol. 1990; 215(3): 403–410.
25. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximumlikelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011; 28(10): 2731–2739.
26. Palevich N, Britton C, Kamenetzky L, Mitreva M, de Moraes MM, Bennuru S, et al. Tackling hypotheticals in helminth genomes. Trends Parasitol. 2018; 34: 179–83.
27. Ndosi BA, Lee D, Bia MM, et al. Morphometry and Molecular Identification of Haemonchus Cobb, 1898 (Trichostrongylidae: Nematoda) Isolates from Small Ruminants in Tanzania Based on Mitochondrial cox 1 and rRNA-ITS genes. J Parasitol Res. 2023; 2023:1923804.
28. Mochizuki R, Endoh D, Onuma M, Fukumoto S. PCR-based species-specific amplification of ITS of Mecistocirrus digitatus and its application in identification of GI nematode eggs in bovine faeces. J Vet Med Sci. 2006; 68(4): 345-351.
29. Food and Agricultural Organization (FAO). “Distribution and impact of helminth diseases of livestock in developing countries,” in FAO Corporate Document Repository— Agriculture and Consumer Protection. 2000.
30. Zarlenga DS, Chute MB, Martin A, Kapel CMO. A multiplex PCR for unequivocal differentiation of all encapsulated and non-encapsulated genotypes of Trichinella. Int J Parasitol. 1999; 29(11): 1859-1867.
31. Ademola IO, Krücken J, Ramünke S, Demeler J, Von G, Himmelstjerna S. Absence of detectable benzimidazole resistance associated alleles in Haemonchus placei in cattle in Nigeria revealed by pyrosequencing of β-tubulin isotype 1. Parasitol Res. 2015;114(5):1997-2001.
32. Rocha RA, Rocha R, Bresciani Ket al. Sheep and cattle grazing alternately: Nematode parasitism and pasture decontamination. Small Rumin Res. 2008;75(2-3):135-143.
33. Nascimento AA, Bonuti MR, Mapeli EB, et al. Natural infections in cervids (Mammalia: Cervidae) from the states of Mato Grosso do Sul and São Paulo by Trichostrongyloidea nematodes. Braz J Vet Res Anim Sci. 1927;37(2):153-158.
34. Zarlenga DS, Stringfellow F, Nobary M, Lichtenfels JR. Cloning and characterization of ribosomal RNA genes from three species of Haemonchus (Nematoda: Tric-hostrongyloidea) and identification of PCR primers for rapid differentiation. Exp Parasitol. 1994; 78(1): 28-36.
35. Palevich N, Maclean PH, Choi YJ, Mitreva M. Characterization of the complete mitochondrial genomes of two sibling species of parasitic roundworms, Haemonchus contortus and Teladorsagia circumcincta. Front Genet. 2020;11: 573395.
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| Issue | Vol 19 No 4 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v19i4.17165 | |
| Keywords | ||
| Cattle Haemonchus placei Multiple sequence alignment Nigeria | ||
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How to Cite
1.
Banwo O, Jeremiah O, Adesina R, Adeyemo A, Fagbohun O. Molecular Detection and Characterization of Haemonchus spp. in Cattle in Nigeria. Iran J Parasitol. 2024;19(4):448-456.
