Epidemiology, morphometry, and phylogenetic analysis of Haemonchus contortus in small ruminants of Malakand division, Pakistan

M. I. Khan; W. Khan; A. Akbar; A. Ullah; A. Ali; A. U. Khan; P. R. De Los Ríos-Escalante

doi:10.2478/helm-2025-0023

Figures & Tables

The study area map displays the geographical distribution of sampling locations in the Malakand Division of Khyber Pakhtunkhwa, Pakistan. It includes a national context map (country map), a provincial focus map (province map), and detailed district boundaries within the study area, highlighting Dir Upper, Dir Lower, Swat, Malakand, Shangla, and Buner. The map was designed and modified by the authors.

Morphological features of Haemonchus contortus.
(A). Egg containing morula, showing measured length and width (μm); (B). Male bursa highlighting spicules and caudal rays, with spicule length (mm); (C). Adult male and Female H. contortus aligned on a calibrated scale to show total body length (mm); (D). Posterior end of female showing anus and measured tail length (mm); (E). Vulvar flap of female Haemonchus; (F). Cervical papillae with recorded length (mm); (G). Oesophagus of H. contortus showing total length (mm) — Morphological features of Haemonchus contortus. (A). Egg containing morula, showing measured length and width (μm); (B). Male bursa highlighting spicules and caudal rays, with spicule length (mm); (C). Adult male and Female H. contortus aligned on a calibrated scale to show total body length (mm); (D). Posterior end of female showing anus and measured tail length (mm); (E). Vulvar flap of female Haemonchus; (F). Cervical papillae with recorded length (mm); (G). Oesophagus of H. contortus showing total length (mm)

Agarose gel electrophoresis showing PCR amplification of COX region of H. contortus.

Phylogenetic tree of H. contortus in Goats and Sheep using Cox-1 marker. The constructed tree illustrates the genetic links between the isolates of the current study and reference sequences from other areas. Bootstrap values are given at branch points representing the group’s confidence levels.

Overall prevalence of H_ contortus in goats and sheep in Malakand division_

Host	Sample	Positive	Percentage	P-value
Goats	398	96	24.12	0.51379^ns
Sheep	480	125	26.04
Total	878	221	25.17

Morphometric measurement (in mm) of H_ contortus isolated from goats and sheep_

Particulars	Male worms (n= 50)	Female worms (n= 76)
Body length (mm)	11.9 – 13.8	15 – 23.8
Body width (mm)	0.23 – 0.38	0.29 – 0.38
Cervical papillae (mm)	0.28 – 0.38	0.24 – 0.42
Spicule length (mm)	0.44 – 0.78	–
Length of esophagus (mm)	0.61 – 0.92	0.72 – 1.22
Gubernaculum length (mm)	0.09 – 0.18	–
Distance from vulva to posterior end (mm)	–	0.68 – 0.82
Distance from Anus to posterior end (mm)	–	0.29 – 0.35

Epidemiological spectrum of Haemochus contortus among sheep and goat populations in Pakistan_

Study area	Host	Sample type	Method	Samples	Positive	Prev. %	QAS	Reference
KPK	Goat & Sheep	Fecal	Mic & Mol	180	46	25.55	5	Ahmad et al., 2024
Sindh	Goat & Sheep	Fecal	Mic & Mol	800	88	11	4	Memon et al., 2024
KPK	Goat & Sheep	Fecal	Mic, Floa & Sed	1340	312	23.28	4	Irshad et al., 2023
KPK	Goat & Sheep	Fecal	Floa & MM	400	57	14.25	5	Khan et al., 2022
KPK	Goat & Sheep	Blood & Fecal	Mic	300	56	18.66	2	Ali et al., 2022
KPK	Goat & Sheep	Fecal	Mic	480	63	13.12	3	Aimen et al., 2022
KPK	Goat & Sheep	Fecal	Mic	315	71	22.53	3	Ruhoollah et al., 2021
Punjab	Goat & Sheep	Worm	Mic	200	105	52.2	3	Bibi et al., 2017
KPK	Sheep	Fecal	Floa, sed & MM	300	120	40	4	Jamil et al., 2016
Punjab	Goat & Sheep	Fecal	Mic & EPG	646	178	27.55	5	Qasim et al., 2016
KPK	Goats	Fecal	Mic	150	22	14.66	4	Nabi et al., 2014
Punjab	Goat & Sheep	Fecal	Floa & Sed	1000	133	13.3	4	Raza et al., 2014
Punjab	Goat	Fecal	Floa & Sed	100	20	20	3	Ayaz et al., 2013
Punjab	Goat & Sheep	Fecal	Floa, Sed & MM	480	137	28.54	4	Khalid et al., 2013
Balochistan	Sheep	Fecal	McMaster	1200	125	10.42	2	Razzaq et al., 2013
KPK	Sheep	Fecal	Mic & Floa	356	190	53.37	3	Ullah et al., 2013
Punjab	Sheep	Fecal	Mic, Floa & Sed	523	34	6.5	4	Lashari & Tasawar, 2011
Sindh	Goat	Fecal	Mic	1065	156	14.65	3	Akhter et al., 2011
Punjab	Sheep	Fecal	MM	333	259	77.7	3	Tasawar et al., 2010
Punjab	Goat & Sheep	Worm	K/Iden	4740	1604	33.83	1	Raza et al., 2009
Punjab	Goat & Sheep	Fecal	Mic, Floa & Sed	400	225	56.25	1	Gadahi et al., 2009
Punjab	Goat	Fecal	Mic, Floa & MM	300	84	28	2	Ijaz et al., 2008
Punjab	Sheep	Worm	Mic & K/Iden	960	591	61.5	3	Lateef et al., 2005
Punjab	Sheep	Fecal	Mic	10000	5477	54.77	2	Jabeen et al., 2000
Islamabad	Sheep	Fecal	Floa & MM	62	54	87	0	Sajid et al., 2000

Prevalence of Haemonchus contortus in small ruminants concerning various risk factors in Malakand division_

Risk Factors	Variables	Total samples	Total positive cases	Total prevalence (%)	Goats Positive (%)	Sheep Positive (%)	P-value
Gender	Male	368	85	23.1	22.62	23.50	0.2292^ns
Gender	Female	510	136	26.7	25.22	27.86	0.2292^ns
Age	<1 years	235	66	28.1	29.25	27.13	0.0391*
	1-5 years	325	66	20.3	17.36	22.65
	>5 years	318	89	28	27.03	28.82
Area	Swat	164	43	26.2	23.19	28.42	0.9815^ns
	Shangla	135	33	24.4	23.61	25.40
	Dir (L)	156	41	26.3	25.00	27.50
	Dir (U)	130	30	23.1	23.21	22.97
	Malakand	160	42	26.2	25.76	26.60
	Buner	133	32	24.1	23.73	24.32
Grazing system	Grazing	465	126	27.1	26.19	27.84	0.1629^ns
Grazing system	Stall-fed	413	95	23	21.81	24.00	0.1629^ns
Health condition	Healthy	435	86	19.8	18.78	20.59	0.0003*
	Moderate	338	97	28.7	26.97	30.11
	Poor	105	38	36.2	36.73	35.71
Drinking water source	Open	285	81	24.4	28.46	28.39	0.1729^ns
	Bore	187	40	21.4	20.45	22.22
	Stream	211	58	27.5	25.56	28.93
	Tap water	195	42	21.5	20.00	22.86
Season	Autumn	205	48	23.4	21.98	24.56	0.5016^ns
	Winter	190	42	22.1	20.51	23.21
	Spring	223	59	26.5	25.23	27.59
	Summer	260	72	27.7	27.05	28.26
Treatment	Treated	290	50	17.2	16.55	17.88	0.0004*
Treatment	Untreated	588	171	29.1	28.19	29.79	0.0004*

Morphometric characteristics of H_ contortus egg sizes (in μm)_

Shape	Statistical index	Length	Width
		μm	μm
Oval	Average	80.98	45.05
	Maximum	99	61
	Minimum	63	36
	Standard error	1.6016	1.43
	Standard deviation	9.4749	8.49
	Sample variance	89.7754	72.14
	N	35	35

Epidemiology, morphometry, and phylogenetic analysis of Haemonchus contortus in small ruminants of Malakand division, Pakistan

Figures & Tables

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Overall prevalence of H_ contortus in goats and sheep in Malakand division_

Morphometric measurement (in mm) of H_ contortus isolated from goats and sheep_

Epidemiological spectrum of Haemochus contortus among sheep and goat populations in Pakistan_

Prevalence of Haemonchus contortus in small ruminants concerning various risk factors in Malakand division_

Morphometric characteristics of H_ contortus egg sizes (in μm)_

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