Prevalence of gastrointestinal parasitic infection in cows and buffaloes in Lower Dir, Khyber Pakhtunkhwa, Pakistan

Abstract Gastrointestinal (GI) Parasitic infection is a hot issue for cattle management. There is variation of GI parasites effects in sex, age of cattle, drinking water condition, nutrition, and severity of infection. Studies on prevalence of GI parasites among cattle population in Dir Lower are lacking. A total of 40 farms were selected randomly in six tehsil namely Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda ,Tehsil Lalqala, Tehsil Khall. Freshly cattle fecal samples were collected randomly from the selected farms during March 2018 till December 2018. Out of 314 buffaloes and cattle examined 58.59% (184/314) were positive for eggs, cyst/oocyst of one or more species of GI Parasites. The prevalence of parasitic infection was higher in Buffaloes 63.55% (75/118) as compared to Cow 55.61% (109/196) but the difference was not significant (p>0.05) Entamoeba,spp, Moniezia spp, Haemonchus spp and Coccidian spp were found in this study. The non-treated animals indicated the highest percentage of infection in cow 57.71% (101/175) and buffalo 68.13% (62/91).GI parasite prevalence in female animal were higher female cow 62.58% (87/139) and female buffalo 77.33% (58/75) as compared to male. But the difference is non-significant (p> 0.05) Yearling calves had the lower rate of GI parasitic infection than adults. Future investigations are necessary to evaluate the economic loss due to GI parasites in cattle’s.


Introduction
The agriculture sector is has a key role in developing the economy of a country.Pakistan is considered an agricultural country, but with repaid decline in the performance of homes of the host (Cuomo et al., 2009).Large ruminants are pretentious with dissimilar types of vermin (Coop and Kyriazakis, 1999), while in some case are deadly because of the parasites types.
Because of unhygienic environment, improper care, close interaction with septic animals and extreme climate sheep and goat become infected with large variability of gastrointestinal parasites (Gadahi et al., 2009.).Most of ruminants badly are affected by helminthiasis, which cause biochemical and hematological disturbance, anorexia, poor reproductive performance, weight loss and decreased animal confrontation to diseases that can lead to death or even become the cause of lots of mortality among the ruminant and lead to a great economic loss (Ngategize et al., 1993).It is reported that reduction occurred in farmer profit up to 16% and decrease in weight up to 50% is because of gastrointestinal parasitic infections.The current study was conducted to investigate the prevalence of gastrointestinal parasitic infection as to find out which parasites might be harboured by the large ruminants in Lower Dir district, Pakistan.

Study Area
This study was conducted at District Dir lower Khyber PukhtunKhwa, Pakistan.The district lies in the northern highlands of KP at a latitude 34.35, longitude 71.85 and altitude of 823.The average annual temperature is 16°C and average annual rainfall is 1186 mm recorded.The relative humidity varies from 70 to 81% in rainy season and in dry season from 40 to 50% humidity is recorded.

Study Population
The study population consisted of all buffalos and cows in the study area.

Collection
Fecal samples were collected directly from the rectum of buffalos and cows randomly, using gloved fingers, put in clean sampling plastic bottles containing 10% formalin and giving sampling number.A questionnaire was also designed which included the following parameters: animal category, date, gender, area, age, season, grazing system, health status, condition of farm/household, treatment, socio-economic status of the farmer and source of drinking water.Samples were brought to the Laboratory of Parasitology in the Department of Zoology, University of Malakand for further analysis.

Laboratory tools
Normal saline (0.85%), formalin, sugar, cotton, refrigerator, cover slip, centrifuge machine, slides, beaker, cello tape, glass rod, stick, measuring cylinder, needle, Pakistan.About 20 percent GDP and 46 percent of labor are engaged in this sector to earn live hood.About 62 percent of people of Pakistan live in rural areas who are directly or indirectly linked with agriculture (Raza et al., 2012).
The role of parasitism is very much significant in most of the agro-ecological regions and a major threat to worldwide livestock economy (Vercruysse and Claerebout, 2001).Helminths infection are considered one of the chief restraint to the production of livestock all over the subtropics and tropic region (Githiori et al., 2004).It reduces the efficiency and productivity cause of high economic losses (Sykes, 1994) and increase mortality which effect the income of farming communities.
An efficient, effective and good strategy is necessary for fruitful implementation and formulation of gastrointestinal parasitic infection control.A periodic surveillance is required for the frequency of gastrointestinal parasitic infections in a particular environment and their association to risk factors which effect the transmission of parasites.From various parts in the world about 0.72 to 84.1% gastrointestinal parasites prevalence has been reported (Bundy et al., 1983;Khan et al., 2010).
Pastoral grazing system is the main cause of gastrointestinal parasitic infections.Many Gastrointestinal parasitic species have developed resistance to antigastrointestinal parasitic drugs (Waller, 1994;Min et al., 2005).Due to Gastrointestinal parasitic infection the productivity of animals has reduced and affected the animal performance also (Waller, 1999).Lower growth rate, impaired tissue deposition, productivity and feed intake depress is caused by subclinical gastrointestinal parasitic infections (Sykes, 1994).In young animals mortality may exceed 40% while losses in weight occur 6-13 kilogram per year in per animal (Waller, 1999;Hawkins, 1993).
Buffaloes and cows are not only the sources of animal proteins, but other products which are made from their fetch are very much important for the use of man.Trematode parasites cause secondary infections as well as weakness, watery diarrhea, weight loss, reduced product quality and decrease in milk production also (Solusby, 1982;Gupta et al., 1978).Enormous economic losses occur due to gastrointestinal parasitic infections.Abdominal vermin are parasites which can damage the gastrointestinal walls of human as well as other animals (Loukopoulos et al., 2007).The gastrointestinal parasites are majour health problems to the health of small ruminants in Upper Dir district, Pakistan (Ruhoollah et al., 2023).Similarly gastro-intestinal parasites are a serous problem in cattle and buffaloes in the Loer Dir, Pakistan (Khan et al., 2021).
They live through all the physique, but favour the abdominal wall (Coop and Holmes, 1996).Protozoans have the ability that can directly infect if they out from the abdomen into the environment hence the helminthic needed the period of maturation in the soil while other have some extant of intermediate host for the (Arcari et al., 2000).The most constructive places for abdominal parasites are the small part of stomach, ileum, cecum and big intestine (Cuomo et al., 2009).For the endure or replicate in the intestinal tract the vermin have to familiarise with incessant physical changes comparative to the nourishing volumetric flask, gloves, Electronic weigh machine, mask, tea strainer, microscope, questioner, Pasteur pipette, rack, motor and pestle were the laboratory tool.

Laboratory methods
In the laboratory the samples were examined for the detection of gastrointestinal parasites eggs or cyst on the basis of morphology using the following methods.

Normal saline solution
A solution of saline wet mount was made in which a 2 gram of faces were mixed within drop of saline upon a glass slide.Microscope was used for smear examination.

Concentration techniques 2.4.2.1. Floatation and centrifugation
This procedure is very popular and widely used for spotting eggs of nematodes and cestodes.As their eggs are lighter and small, they can float in the flotation liquid.Almost 3 gm of stool sample was taken in a beaker and 42ml of water was added with the help of motor and pestle, sample was grinded lightly and clarified with a tea filter.
The samples which were filtered and poured into plastic tube of 15ml and were centrifuged at 1000 rpm for 5 minutes.The plastic tube was taken out from the centrifuge and with the help of pipette the upper part of water was poured.Then again the tube was filled with NaCl solution and centrifuged at 1000 rpm for 5-7 minutes.NaCl solution was added so that the tube may filled to the tip of the tube.Cover slip was put on top of the tube, so that the Sedum chloride trace the cover slip for few minutes and after few minutes cover slip will be put on a slide and was examined at 10X (Khan et al., 2018).

Sedimentation technique
This procedure is mostly used for spotting trematodes eggs because the trematodes eggs are bit denser than the other eggs .Havier egg settle down in bottom of test tube when its was centrifuged with sodium chloride solution.
With the help of pipette, a drop of deposited material was took on the slide.On it, a drop sodium chloride (NaCl) was added and examined under a microscope at 4Xand 10X (Solusby, 1982).

Statistical analysis
Data was analyzed by used of Graph Pad version 5. Comparative prevalence of various gastrointestinal (GI) parasites group or species was calculated by following formula.
Prevalence (%) = (Number of positive samples / Total number of samples examined) x 100

Discussion
Cows and buffaloes play an important role in the economy of Pakistan in view of elevation of poverty, production of milk owl, leather, and meat.But the gastrointestinal parasites cause huge morbidity and mortality in these animals, affect the economic condition of the country.A handsome amount of money is also spent on the treatment of gastrointestinal parasites of these animals.The current study was conducted in district Lower Dir to find out the gastrointestinal parasites of Cows and buffalos because less attention has been paid to the screening of cows and buffalo parasites.
In present study the overall prevalence occurred 58.59% with cows 55.61% and buffalos 63.55% .Our finding is nearly similar and in line with the result of (Raza et al.,2012 (51%) and buffaloes (47%) (Jittapalapong et al., 2011 ) cows 46.6% Dula, 2018 cows 40% but our result are less from the finding of (Marskole et al., 2016 ) Baffaloes 73.33% and Cows 75% were found.It may be due to differences in environmental status of the study area and methodology.
High prevalence was observed in female cows 62.58% while more occurrence also in female buffaloes 77.33%. in this regard our finding is nearly similar in line with the result of Dula, 2018 found 39.9% in male cows and 41.6% prevalence found in female cows but our result are less from the finding of (Raza et al., 2007) which may be during samples collection it was observed that more peoples keep female cows and buffalos for their daily life requirement like milk, and other dairy products.It may be due sensitive immune system of females animals during pregnancy and hormonal differences.
Regarding age, adult cows were highly affected by GI parasites 56.12% while more occurrence where also occurred in adult buffaloes 73.8%.Our finding is nearly  Qureshi and Tanveer (2009); Telila et al. (2014).On the contrary, Ahmed et al. (2017) stated that the younger animals are more susceptible than adult animals.The more occurrence in adult animals maybe due to the weakness of immune system, and regular exposure to infected animals and contact with infected other animals.Prevalence in treated cows was 38.09% and untreated cows 57.71% as well as, in treated buffaloes the occurrence was found 48.14% while in untreated buffalo 68.13%.Our finding is closely similar and in line with the result of Gunathilaka et al. (2018) cows untreated 46.67% and treated cows 15% and untreated Buffalos 48.56% and treated Buffalos 17.78%, but our results are different from the finding of Githiori et al. (2004).The high occurrence in untreated cows and buffalos were noted as compared to treated animals .The high prevalence rates in untreated animals may be due to untreated and lacking the used of medicine, un awareness in farmers about the use of anti-helminthic drugs to control GI parasites in animals.

Table 1 .
Overall prevalence of Intestinal parasites in cows and buffalos in Lower Dir, Pakistan.

Table 4 .
Treatment wise prevalence of GI parasites in cows and buffalos in District Lower Dir, Pakistan.

Table 2 .
Gender wise prevalence of GI parasites in cows and buffalos in district Lower Dir Pakistan.

Table 3 .
Age wise prevalence of Intestinal parasites in cows and buffalos in district Lower Dir Pakistan.Gastrointestinal Parasitic Infections in Cows and Buffaloes similar and in line with the result of Dula, 2018 cows young 38.3% while adult cows 41.9% but our result are less from the finding of Marskole et al., 2016 adult cows 77.50% and Adult Buffaloes 75% as compared to young which are 56%in cattle and 62.59% in young Buffalos.The increase in the prevalence of GI parasites with the age has also been reported by