Table 1.
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′ |
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.
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
Fig. 4.
Knobbed/button type vulvar flap of H. contortus female.
Fig. 5.
Smooth type vulvar flap of H. contortus female
Table 2.
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 |
[i] Note: p value ≤0.05- Significant; p value > 0.05- Non-significant
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.
Table 3.
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 |
[i] 1. 003-Navsari-Male, 2. 001-Navsari-Female, 3. AF044927.1-U.S.A., 4. EU084691.1-U.S.A., 5. JN590059.1-Russia, 6. KJ857556.1-Kolkata, 7. KJ857558.1-Mukteswar, 8. KJ938047.1-Chennai, 9. KP760874.1-Kenya and 10. KX534106.1-China
Table 5.
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 |
[i] 1. 002-Navsari-Female, 2. 004-Navsari-Male Navsari isolate, 3. EU084691.1-U.S.A., 4. JQ342246.1-Brazil, 5. KJ857556.1-Kolkata, 6. KJ938047.1-Chennai, 7. KP101383.1-Thailand, 8. KP760874.1-Kenya, 9. KX534106.1-China, 10. LC430925.1-Nigeria, 11. MH481597.1-Australia, 12. MT645506.1-Bangladesh and 13. X78803.1-Australia
Fig. 7.
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.