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. 2023 Sep 22;60(2):175-188.
doi: 10.2478/helm-2023-0020. eCollection 2023 Jun.

Morphological and molecular characterization of Haemonchus contortus isolated from the small ruminants of south Gujarat, India

B Das  1 , N Kumar  1 , J B Solanki  1 , M M Jadav  1 , I H Kalyani  2
Affiliations

Morphological and molecular characterization of Haemonchus contortus isolated from the small ruminants of south Gujarat, India

B Das et al. Helminthologia. .

Abstract

The successful design of strategic control measures against the blood-sucking gastrointestinal nematode, Haemonchus contortus in small ruminants can be facilitated by revealing its general features from morphology to the molecular level. In the south Gujarat region of India, a total of 2408 H. contortus were collected from 84 slaughtered sheep's abomasum, consisting of 347 males and 2061 females (1:6 ratio) (p<0.05). Furthermore, 726 H. contortus were collected from 61 goats, comprising 145 males and 581 females (1:4 ratio) (p<0.05). The male worms were approximately 12±0.06 mm long, while female worms were about 20±0.09 mm long. The vulvar morphotypes of the female worms were found to be 17.7% linguiform, 76.6 % knobbed/button (p<0.05), and 5.7 % smooth type, demonstrating common features of H. contortus. The nucleotide sequences of the Internal Transcribed Spacer 1 (ITS-1) of 165 bp or ITS-2 plus of 256 bp were aligned, and it was found that the genotypes of male and female specimens of either sheep or goat origin were identical, with a 100 % match. The present isolates shared >95 % and >94 % homology with published sequences of ITS-1 and ITS-2 plus of H. contortus, respectively, with more nucleotide transitions than transversions in the aligned sequences. The reconstructed phylogram of either ITS-1 or ITS-2 plus revealed two major clades, one for H. contortus and another for other nematodes, with Haemonchus placei showing its proximity with the clade of H. contortus. The study established the role of morphological and molecular features in identifying and differentiating H. contortus parasite at the local level.

Keywords: Haemonchus contortus; ITS; Molecular; PCR; Small ruminants; Vulvar morphology.

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Conflict of interest statement

Conflict of Interest The authors declare that they have no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Bursa of male H. contortus showing Y shaped dorsal ray and each spicule with a single barb at its distal tip.
Fig. 2.
Fig. 2.
Intertwining of red intestine and white ovary of H. contortus female
Fig. 3.
Fig. 3.
a: Linguiform type B vulvar flap of H. contortus female; b: Linguiform type I vulvar flap of H. contortus female
Fig. 4.
Fig. 4.
Knobbed/button type vulvar flap of H. contortus female.
Fig. 5.
Fig. 5.
Smooth type vulvar flap of H. contortus female
Fig. 6.
Fig. 6.
PCR amplification of ITS-1 of 198 bp (lane 1 and 3) and ITS-2 of 350 bp (lane 2 and 4) genotype sequence of Navsari isolates of H. contortus. Lane M: 100 bp DNA ladder.
Fig. 7.
Fig. 7.
Phylogenetic relationship of Haemonchus along with some other important nematodes based on ITS-1 sequences.
Fig. 8.
Fig. 8.
Phylogenetic relationship of Haemonchus along with some other important nematodes based on ITS-2 plus sequences.
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