Diagnosis of Oestrus ovis infestation in sheep by PCR and enzyme-linked immunosorbent assay

Bello, Hornblenda Joaquina Silva; Lins, José Gabriel Gonçalves; Silva, Naiara Mirelly Marinho da; Albuquerque, Ana Cláudia Alexandre de; Amarante, Mônica Regina Vendrame; Neto, Vitoldo Antônio Kozlowski; Amarante, Alessandro Francisco Talamini do

doi:10.1016/j.vetpar.2022.109789

Cited by 4 publications

(2 citation statements)

References 38 publications

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“…Moxidectin is a macrocyclic lactone and belongs to the same drug class as ivermectin. Phenotypically, resistance to moxidectin typically confers resistance to ivermectin; however, resistance to moxidectin may occur independently of ivermectin resistance in C. elegans (37), a phenomenon also found in a recent field study of gastrointestinal nematodes on sheep farms (38). A single peak on chromosome 3 (peak range: 15-20 Mb) found only in the MTci7:MTci2 comparison and not in the other ivermectin-resistant isolate comparisons is suggestive of a moxidectin-specific response.…”

Section: Genetic Variation Associated With Individual Drug Classesmentioning

confidence: 61%

Chromosomal genome assembly resolves drug resistance loci in the parasitic nematode Teladorsagia circumcincta

McIntyre,

Morrison,

Maitland

et al. 2024

Preprint

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The parasitic nematode Teladorsagia circumcincta is one of the most important pathogens of sheep and goat farming in temperate climates worldwide and can rapidly evolve resistance to drugs used to control it. To understand the genetics of drug resistance, we have generated a highly contiguous genome assembly for the UK T. circumcincta isolate MTci2. Assembly using PacBio long-reads and Hi-C long-molecule scaffolding together with manual curation resulted in a 573 Mb assembly (N50 = 84 Mb, n = 1,286) with five autosomal and one sex-linked chromosomal-scale scaffolds consistent with its karyotype. The genome resource was further improved via annotation of 22,948 genes, with manual curation of over 3,200 of these, resulting in a robust and near complete resource (96.3% complete protein BUSCOs) to support basic and applied research on this important veterinary pathogen. Genome-wide analyses of drug resistance, combining evidence from three distinct experiments, identified selection around known candidate genes for benzimidazole, levamisole and ivermectin resistance, as well as novel regions associated with ivermectin and moxidectin resistance. These insights into contemporary and historic genetic selection further emphasise the importance of contiguous genome assemblies in interpreting genome-wide genetic variation associated with drug resistance and identified key loci to prioritise in developing diagnostic markers of anthelmintic resistance to support parasite control.

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Section: Genetic Variation Associated With Individual Drug Classesmentioning

confidence: 61%

Chromosomal genome assembly resolves drug resistance loci in the parasitic nematode Teladorsagia circumcincta

McIntyre,

Morrison,

Maitland

et al. 2024

Preprint

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“…The grazing area of sheep is invaded by grass species including Kikuyu ( Pennisetum clandestinum) and speargrass In addition, the oral administration of commercial preparation of Ivermectin (HITEK R : Each ml contains 0.800 mg of Ivermectin : Dose -25 ml/100 Kg body weight) for the treatment of gastrointestinal parasitism among sheep at SRRC, Mannavanur, is a periodical practice and this could be the reason for the absence of typical clinical cases of Oestrosis among sheep in the present study. Bello et al 2022) reported that in sheep, oral drenching of with Closantel at a dose rate 10 mg/kg and subcutaneous injection with ivermectin at a dose rate of 0.2 mg/kg could be effective in the control of oestrosis.…”

Section: Resultsmentioning

confidence: 99%

First report on molecular characterization of Oestrus ovis in sheep from India

Nagarajan,

Kanagarajadurai,

Pachaiyappan

et al. 2024

J Parasit Dis

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The objective of the present study is to characterize the dipteran larvae species infesting the sheep being maintained at SRRC, Mannavanur, by means of COI gene based PCR. During the last week of May 2021,post mortem examination of the skull of an Avikalin male sheep (20 months old) revealed the presence of larvae in its nasal sinuses. The larvae were washed in PBS (pH 7.2) and preserved in 70% alcohol. Total genomic DNA was isolated from the larvae using an initial step of grinding with liquid Nitrogen in a sterile mortar and pestle. Using the isolated genomic DNA from the larvae as a template, Cytochrome c oxidase subunit I (COI) gene based PCR was employed using the primers designed based on the COI gene of reference isolate of Oestrus ovis available in the GenBank. Full length COI gene (1534 bp) gene of Oestrus ovis in sheep from South India was targeted in the PCR experiment. The pTZ57R/T vector was used for the cloning of the PCR ampli ed fragment and the con rmed recombinant plasmid was subjected to sequencing experiments. In addition to morphological examination, based on COI gene based PCR, eventual sequencing experiments and BLAST analysis, it was con rmed that the larvae in the nasal sinuses of sheep from South India were Oestrus ovis. The South Indian isolate of Oestrus ovis is sharing 100% sequence identity both at nucleotide and amino acid levels with that of O.ovis from Spain. The North Indian isolate of O.ovis (from Jammu) exhibited 92 & and 99% identity at respective nucleotide and amino acid levels with South Indian isolate. With other members of the subfamily Oestrinae, the share of per cent nucleotide and amino acid identities of South Indian O.ovis ranged from 85-86% and 95-96%, respectively. O.ovis from South India was grouped with the other members of Oestrinae from different geographical areas of the globe in the analysis of phylogenetic tree based on COI amino acid sequences. Based on the research ndings, it is concluded that Oestrus ovis is the dipteran species infesting the sheep at Mannavanur, Tamil Nadu, India. To our knowledge, this is the rst report on full length nucleotide sequences of COI gene of O.ovis in sheep from Indian subcontinent.

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First report on Molecular Characterization of Oesturs ovis in sheep from India

Nagarajan

Kanagarajadurai

Pachaiyappan

et al. 2023

Preprint

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Purpose Upon post mortem examination of sheep (from SRRC, Mannavanur, South India) having the history of natural death, the nasal sinuses of the animals contained dipteran larvae. The objective of the present study is to characterize the dipteran larvae species infesting the sheep being maintained at SRRC, Mannavanur, by means of COI gene based PCR. Methods During the last week of May 2021, an Avikalin male sheep (20 months old) died suddenly. Upon Post mortem examination, the skull of the dead sheep was having of larvae in the nasal sinuses. The larvae were washed in PBS (pH 7.2) and preserved in 70 % alcohol. The larvae taken out of 70 % alcohol were thoroughly ground to make a fine powder using liquid Nitrogen in a sterile mortar and pestle. The maggot in powder form was then used for the isolation of total genomic DNA isolation. Using the isolated genomic DNA from the larvae as a template, Cytochrome c oxidase subunit I (COI) gene based PCR was employed using the primers designed based on the COI gene of reference isolate of Oestrus ovis available in the GenBank. Full length COI gene (1534 bp) gene of Oestrus ovis in sheep from South India was targeted in the PCR experiment. The pTZ57R/T vector was used for the cloning of the PCR amplified fragment and the confirmed recombinant plasmid was subjected to sequencing experiments. The resultant nucleotide sequences of COI gene of the O.ovis in sheep from South India was analysed with that of 29 dipteran species from different different geographical areas of the world (available in GenBank) by using standard bioinformatics tools. Results In addition to morphological examination, based on COI gene based PCR, eventual sequencing experiments and BLAST analysis, it was confirmed that the larvae in the nasal sinuses of sheep from South India were Oestrus ovis. The South Indian isolate of Oestrus ovis is sharing 100% sequence identity both at nucleotide and amino acid levels with that of O.ovis from Spain. The North Indian isolate of O.ovis (from Jammu) exhibited 92 & and 99 % identity at respective nucleotide and amino acid levels with South Indian isolate. With other members of the subfamily Oestrinae, the share of per cent nucleotide and amino acid identities of South Indian O.ovis ranged from 85-86 % and 95-96 %, respectively. O.ovis from South India was grouped with the other members of Oestrinae from different geographical areas of the globe in the analysis of phylogenetic tree based on COI amino acid sequences. ConclusionBased on the research findings, it is concluded that Oestrus ovis is the dipteran species infesting the sheep at Mannavanur, Tamil Nadu, India. To our knowledge, this is the first report on full length nucleotide sequences of COI gene of O.ovis in sheep from Indian subcontinent. An investigation on the report of human ocular infections and the accidental hosts for O ovis in the area of the present study has to be explored.

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Diagnosis of Oestrus ovis infestation in sheep by PCR and enzyme-linked immunosorbent assay

Cited by 4 publications

References 38 publications

Chromosomal genome assembly resolves drug resistance loci in the parasitic nematode Teladorsagia circumcincta

Chromosomal genome assembly resolves drug resistance loci in the parasitic nematode Teladorsagia circumcincta

First report on molecular characterization of Oestrus ovis in sheep from India

First report on Molecular Characterization of Oesturs ovis in sheep from India

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