Prevalence and taxonomic identification of hard ticks (Ixodidea) found in livestock of Harnai District, Balochistan, Pakistan

Ticks cause considerable damage to the livestock in tropical and subtropical areas. Because of climatic and environmental changes, the frequencies of tick-borne infections are increasing day by day. This research trial displays an outline about the different species of hard ticks (Family; Ixodidae) found in sheep, goats, cows, buffaloes and camels in District Harnai, Baluchistanon. The study duration of the present research trial was from 1 st December 2018 to 1 st December 2019. Ticks were collected from different parts of the body and examined for species identification. Total 913 ticks were collected from livestock. Most susceptible animal was cow (65%) followed by buffalo (55%), camel (47.5%), sheep (30%) and goats (27.5%). Prevalence with respect to season, it was noted that, in summer season all hosts i.e. cows, buffalo, camels, goats and sheep were found to be highly infested while in winter infestation was low. In the current research trial, the prevalence of tick was high in cattle as compared to other livestock of Harnai District, Baluchistan. Nine species of ticks were identified in the study area as Amblyomma variegatum, Amblyomma heberaeum, Rhipicephalus Boophilus, Hyalomma dromedarii, Rhipicephalus appendiculatus, Rhipicepalus boophilus microplus, Rhipicephalus Boophilus annulatus, Hyalomma anatolicum, and Ixodes rubicundus. The widely spread and most prevalent species of tick were Amblyomma heberaeum and Rhipicephalus appendiculatus, while Ixodes rubicundus was the least prevalent specie. Favorable climatic conditions may be one of the reasons of this high prevalence of ticks in District Harnai.


Introduction
Economy of Pakistan mainly depends on its livestock production as chief provider of food [1]. The domesticated animals are the vital piece of the economy of Pakistan and viewed as the foundation of the rural economy as over 70% of the population lives in rural areas. People rely upon keeping domesticated animals for their survival [2]. Ticks are extremely well known specific group of obligate ectoparasites of birds, reptiles and mmamles. They have blood sucking feeding habit [3]. There are three types of ticks identified as hard ticks, soft ticks and nuttalliella [4]. The differences between male and female ticks are fallowing. The male have totally covered scutum and female covers only a portion of dorsal surface [5]. Ticks are economically responsible for great lasses in cattle farming worldwide [6]. Ticks are the specific type of ectoparasites, which suck blood from warm blooded animals and have a potential to transfer parasites mostly in animals and rarely in humans. These parasites cause serious loses by spreading pathogens among animal populations [7]. Ixodidae family is one of the biggest groups of parasitic ticks in which about 13 genera and 650 species have been recognized [8].
Ticks of Ixodidae family transmit various bacterial, protozoal, rickettsial, spirochaetal and viral pathogens. The ixodid tick species are the most common tick parasites found in ruminants. Ticks have been thought as the critical danger because of their sensitivity and toxicity to animals. Ticks cause considerable damage to the assets of cultivating community particularly in tropical and subtropical areas

Study design
To identify and determined the prevalence of hard ticks in livestock, ticks were collected from upper, lower, anterior and posterior parts of the body and examined for species identification. Tick collection and preservation Livestock were thoroughly inspected for ticks from every part of the body (ear, eye, testis, udder, vulva, tail, memory gland, abdominal and anal area). 30% alcohol was used to preserve ticks, after being separated from the host body.

Tick processing and identification
In lab separate the sample according to their specific features start the process including After boiling for 20-30 min in potassium hydroxide (KOH), the observed ticks were then passed through a series of grading level of alcohol (30%, 50%, 70%, 90% and 100%). Ticks were placed for 2 hours in each level of alcohol then transferred to the higher grade. Staining was done before transferring to 90% grade of alcohol. After 100%, finally ticks were placed in xylol for 5 seconds and then clove oil for few seconds. Ticks were mounted from Canada balsam after placing on slide. After processing all ticks were identified with the help of a microscope by taxonomy keys [10].

Sex determination
On the basis of scutum on the dorsum, female ticks had been separated and all ticks were identified for genus and specie level.

Results
The most susceptible animal was cow (65%) followed by buffalo (55%), camel (47.5%), sheep (30%) and goat (27.5%) as shown in (Table 1). The male and female were observed from every host i.e. cow, buffalo, camel, goat and sheep. It was noted that 48 males and 42 females were found infested (Table 1) (Table 2). In cow 133 ticks were males and females were 179. The total male: female ratio was (1:1.4), which showed dominancy of female ticks over males. in (sheep) 1:1.5,(cows) 1:1.2 and (camels)1:1.9 respectively. Rhipicepalus Boophilus was observed in maximum number as larvae N (10) and nymp N (19) in while Amblyomma herbraeum was detected in highest number as adult N (72) ( Table 3). In summer season all hosts i.e. cows, buffalo, camels, goats and sheep were found to be highly infested while in winter infestation was low. In July high number of ticks were collected, which was followed by August (Table 4), while in February, minimum number of ticks were observed on cattle. This proved a direct association of tick's prevalence with increased temperature. Humidity and temperature may influence tick infestation.   and Rhipicephalus appendiculatus, while Ixodes rubicundus was the least prevalent specie (Table 5). This further indicated that the finding of this tick in the area is in line with its wide spread occurrence in most parts of the country. Hyalommaand Rhicephalus had also been confirmed from elevated and hilly areas and from grasslands areas of Pakistan [14]. The authors examined 813 cattle in Gilgit Baltistan and Khyber Pakhtunkhwa. It was observed that the rate of prevalence of Hyalomma anatolicum and Rhipicephalus microplus was exceedingly high as compare to other species. In another study. Khalil et al. [15]. found Hyalomma as highly prevalentspeciewith a rate of 44.5% prevalence in buffaloes of Jampur District, South Punjab, Pakistan. Esmail & Ahari, [16]; Radfar [17] also stated Hyalomma as dominating specie over other species of ticks. In cow 133 ticks were males and females were 179. The total male: female ratio was (1:1.4), which showed dominancy of female ticks over males. in (sheep) 1:1.5,(cows) 1:1.2 and (camels)1:1.9 respectively. Rhipicepalus Boophilus was observed in maximum number as larvae N (10) and nymp N (19) in while Amblyomma herbraeum was detected in highest number as adult N (72). In summer season all hosts i.e. cows, buffalo, camels, goats and sheep were found to be highly infested while in winter infestation was low. In July high number of ticks were collected, which was followed by August, while in February, minimum number of ticks were observed on cattle. This proved a direct association of tick's prevalence with increased temperature. Humidity and temperature may influence tick infestation [18].  and Rhipicephalus appendiculatus, while Ixodes rubicundus was the least prevalent specie. In general, due to unsatisfactory tick control measures and lack of awareness about effective control strategies, ticks were highly prevalent in cattle in the study area. Favorable climatic conditions may be one of the reason of this high prevalence of ticks in District Harnai. The ixodid tick have been thought as the critical danger to cattle because of their sensitivity and toxicity to animals and accountable for severe financial losses through impairment to hide and by spreading various diseases through blood sucking. Consequently, it is necessary to control tick infestation in cattle to reduce the loss. Use of acaricide could be a good mechanism to minimize tick infestation. Further studies on the impact of environment on tick infestation, other risk factors and as well as effective tick controlling strategies are suggested in the area as per current finding.