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

B. Das
B. Das
, 
N. Kumar
N. Kumar
, 
J. B. Solanki
J. B. Solanki
, 
M. M. Jadav
M. M. Jadav
 and 
I. H. Kalyani
I. H. Kalyani
   | Sep 22, 2023
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Published Online: Sep 22, 2023
Page range: 175 - 188
Received: Nov 29, 2022
Accepted: May 30, 2023
DOI: https://doi.org/10.2478/helm-2023-0020
Keywords
© 2023 B. Das et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Bursa of male H. contortus showing Y shaped dorsal ray and each spicule with a single barb at its distal tip.
Bursa of male H. contortus showing Y shaped dorsal ray and each spicule with a single barb at its distal tip.

Fig. 2.

Intertwining of red intestine and white ovary of H. contortus female
Intertwining of red intestine and white ovary of H. contortus female

Fig. 3.

a: Linguiform type B vulvar flap of H. contortus female; b: Linguiform type I vulvar flap of H. contortus female
a: Linguiform type B vulvar flap of H. contortus female; b: Linguiform type I vulvar flap of H. contortus female

Fig. 4.

Knobbed/button type vulvar flap of H. contortus female.
Knobbed/button type vulvar flap of H. contortus female.

Fig. 5.

Smooth type vulvar flap of H. contortus female
Smooth type vulvar flap of H. contortus female

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.
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.

Phylogenetic relationship of Haemonchus along with some other important nematodes based on ITS-1 sequences.
Phylogenetic relationship of Haemonchus along with some other important nematodes based on ITS-1 sequences.

Fig. 8.

Phylogenetic relationship of Haemonchus along with some other important nematodes based on ITS-2 plus sequences.
Phylogenetic relationship of Haemonchus along with some other important nematodes based on ITS-2 plus sequences.

Multiple alignment percent identity matrix of ITS-1 of H. contortus.

1 2 3 4 5 6 7 8 9
2 100.0
3 97.0 97.0
4 95.8 95.8 98.8
5 96.3 96.3 98.8 98.8
6 96.3 96.3 98.8 98.8 100.0
7 96.3 96.3 98.8 98.8 100.0 100.0
8 96.4 96.4 99.4 99.4 99.4 99.4 99.4
9 95.7 95.7 98.8 98.8 98.1 98.1 98.1 99.4
10 95.2 95.2 98.2 99.4 98.1 98.1 98.1 98.8 98.2

Multiple alignment percent identity matrix of ITS-2 plus of H. contortus.

1 2 3 4 5 6 7 8 9 10 11 12
2 100.0
3 100.0 100.0
4 99.1 99.1 99.0
5 98.4 98.4 98.4 98.4
6 99.5 99.5 99.5 98.4 97.8
7 99.5 99.5 99.5 99.5 97.8 98.9
8 98.0 98.0 98.0 99.0 97.8 97.3 98.5
9 100.0 100.0 100.0 99.0 98.4 99.5 99.5 98.0
10 99.6 99.6 99.5 98.7 97.8 98.9 99.0 97.5 99.5
11 99.5 99.5 99.5 99.5 97.8 98.9 100.0 98.5 99.5 99.0
12 96.5 96.5 96.5 95.5 94.6 95.6 96.0 94.5 96.5 96.0 96.0
13 99.0 99.0 99.0 100.0 98.4 98.4 99.5 99.0 99.0 98.5 99.5 95.5

Distribution pattern of vulvar morphotypes of H. contortus in the small ruminants.

Host Female worms (No.) Vulvar morphotypes p value
Recovered Examined Linguiform Knobbed Smooth
No. % No. % No. %
Goat 2408 2061 342 16.6 1602 77.7 117 5.7 0.045
Sheep 730 581 125 21.5 423 72.8 33 5.7 0.05
Total 3138 2642 467 17.7 2025 76.6 150 5.7 0.05

Primers used for the amplification of the DNA of the nematode.

Name of gene Name of primers Primers sequence Product length References
ITS1 Forward 5′-TATGACATGAGCCGTTCGAG-3′ 198 bp Bandyopadhyay et al., 2011
Reverse 5′-TGATCATTAAGGTTCCCCGA-3
ITS2 plus Forward 5′-ACGTCTGGTTCAGGGTTGTT-3′ 350 bp Stevenson et al., 1995
Reverse 5′-TTAGTTTCTTTTCCTCCGCT-3′
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