Identification of tick species infesting livestock in Dera Ismail Khan Pakistan

Ticks are blood-feeding obligate ectoparasites that belong to order Arachnida with three main families (Ixodidae, Argasidae, and Nuttalleillidae). Ticks transmit viral, bacterial, and zoonotic diseases in humans, and animals (wild and domestic). Ticks directly and indirectly affect the livestock industry which plays an important role in the rural economy of Pakistan. These are the main source of income and a source of food for poor people. The current study was conducted to identify the tick species infesting livestock in Dera Ismail Khan Pakistan. For this purpose, different farms were randomly visited and 130 ticks were collected from 200 scanned animals. The collected ticks were identified to species level using morphological keys. In the current study, four tick species were identified as Rhipicephalus sanguineus, Hyalomma marginatum, Hyalomma anatolicum, and Dermacentor marginatus were identified. The most dominant species were Hy. anatolicum (35.39%) followed by R. sanguineus (30.00%), Hy. marginatum (23.85%) and D. marginatus (10.77%). Hy. anatolicum was the predominant tick species and accounted for 35.39% of the ticks. Tick infestation was highest in buffaloes followed by cows, sheep, and goats. Tail and ear were the most preferable sites for tick infestation than other body parts.

Hyalomma spp. ticks and associated Anaplasma spp. and Ehrlichia spp. on the Iran-Pakistan border

Background Anaplasmosis and ehrlichiosis are tick-borne diseases affecting humans and livestock, particularly in tropical and subtropical regions. Animal husbandry is the main activity of people on the borders of Iran and Pakistan, with thousands of cattle crossing the border each week. Methods PCR and sequencing of the 16S rRNA gene was used to determine the percentage and geographical distribution of the pathogens carried by Hyalomma spp. (n = 306) collected from 126 goats, cattle and camels in the region between November 2017 and late March 2018. Results In total, 1124 hard ticks including 1020 Hyalomma spp. ticks belonging to six species (Hyalomma anatolicum, Hyalomma asiaticum, Hyalomma marginatum, Hyalomma dromedarii, Hyalomma schulzei, and Hyalomma detritum) were found on the borders of Iran and Pakistan, with H. anatolicum being the most prevalent tick species. Anaplasma spp. and/or Ehrlichia spp. DNA was found in 68.3% of the engorged tick specimens (n = 256). Sequencing of a subset (12.6%) of PCR-positive samples revealed Anaplasma ovis, Anaplasma marginale, and Ehrlichia ewingii DNA in 81.8%, 9.1%, and 9.1% of the ticks, respectively. To our knowledge, this is the first report of E. ewingii, an important human pathogen, in Iran. Conclusions Based on molecular analysis, three pathogenic Anaplasmataceae were detected in six Hyalomma spp. parasitizing cattle, goats and camels, confirming the presence of these pathogens along the Iran-Pakistan border. Graphical Abstract

Four Tick-Borne Microorganisms and Their Prevalence in Hyalomma Ticks Collected from Livestock in United Arab Emirates

Ticks and associated tick-borne diseases in livestock remain a major threat to the health of animals and people worldwide. However, in the United Arab Emirates (UAE), very few studies have been conducted on tick-borne microorganisms thus far. The purpose of this cross-sectional DNA-based study was to assess the presence and prevalence of tick-borne Francisella sp., Rickettsia sp., and piroplasmids in ticks infesting livestock, and to estimate their infection rates. A total of 562 tick samples were collected from camels, cows, sheep, and goats in the Emirates of Abu Dhabi, Dubai, and Sharjah from 24 locations. DNA was extracted from ticks and PCR was conducted. We found that Hyalomma dromedarii ticks collected from camels had Francisella sp. (5.81%) and SFG Rickettsia (1.36%), which was 99% similar to Candidatus Rickettsia andeanae and uncultured Rickettsia sp. In addition, Hyalomma anatolicum ticks collected from cows were found to be positive for Theileria annulata (4.55%), whereas H. anatolicum collected from goats were positive for Theileria ovis (10%). The widespread abundance of Francisella of unknown pathogenicity and the presence of Rickettsia are a matter of concern. The discovery of T. ovis from relatively few samples from goats indicates the overall need for more surveillance. Increasing sampling efforts over a wider geographical range within the UAE could reveal the true extent of tick-borne diseases in livestock. Moreover, achieving successful tick-borne disease control requires more research and targeted studies evaluating the pathogenicity and infection rates of many microbial species.

An integrated overview of the bacterial flora composition of Hyalomma anatolicum, the main vector of CCHF

The microbial flora associated with Hyalomma anatolicum ticks was investigated using culture-dependent (CD) and independent (next generation sequencing, NGS) methods. The bacterial profiles of different organs, development stages, sexes, and of host cattle skins were analyzed using the CD method. The egg and female gut microbiota were investigated using NGS. Fourteen distinct bacterial strains were identified using the CD method, of which Bacillus subtilis predominated in eggs, larval guts and in adult female and male guts, suggesting probable transovarial transmission. Bacillus velezensis and B. subtilis were identified in cattle skin and tick samples, suggesting that skin is the origin of tick bacteria. H.anatolicum males harbour lower bacterial diversity and composition than females. The NGS analysis revealed five different bacterial phyla across all samples, Proteobacteria contributing to >95% of the bacteria. In all, 56611sequences were generated representing 6,023 OTUs per female gut and 421 OTUs per egg. Francisellaceae family and Francisella make up the vast majority of the OTUs. Our findings are consistent with interference between Francisella and Rickettsia. The CD method identified bacteria, such B. subtilis that are candidates for vector control intervention approaches such paratransgenesis whereas NGS revealed high Francisella spp. prevalence, indicating that integrated methods are more accurate to characterize microbial community and diversity.

MicroRNA-1 Expression and Function in Hyalomma Anatolicum anatolicum (Acari: Ixodidae) Ticks

MicroRNAs act as mRNA post-transcriptional regulators, playing important roles in cell differentiation, transcriptional regulation, growth, and development. In this study, microRNA expression profiles of Hyalomma anatolicum anatolicum ticks at different developmental stages were detected by high-throughput sequencing and functionally assessed. In total, 2,585,169, 1,252,678, 1,558,217, and 1,155,283 unique reads were obtained from eggs, larvae, nymphs, and adults, respectively, with 42, 46, 45, and 41 conserved microRNAs in these stages, respectively. Using eggs as a control, 48, 43, and 39 microRNAs were upregulated, and 3, 10, and 9 were downregulated in larvae, nymphs, and adults, respectively. MicroRNA-1 (miR-1) was expressed in high abundance throughout Ha. anatolicum development, with an average of nearly one million transcripts, and it is highly conserved among tick species. Quantitative real-time PCR (qPCR) showed that miR-1 expression gradually increased with tick development, reaching the highest level at engorgement. Differential tissue expression was detected, with significantly higher levels in the salivary glands and epidermis than in the midgut. Inhibition assays showed no significant change in body weight or spawning time or amount between experimental and control groups, but there was a significant difference (p < 0.01) in engorgement time. With miR-1 inhibition, ticks displayed obvious deformities during later development. To more fully explain the microRNA mechanism of action, the miR-1 cluster was analyzed according to the target gene; members that jointly act on Hsp60 include miR-5, miR-994, miR-969, and miR-1011. Therefore, microRNAs are critical for normal tick development, and the primary structure of the mature sequence of miR-1 is highly conserved. Nonetheless, different developmental stages and tissues show different expression patterns, with a certain role in prolonging feeding. miR-1, together with other cluster members, regulates mRNA function and may be used as a molecular marker for species origin, evolution analysis, and internal reference gene selection.

Prevalence and intensity of Ixodidae in cattle, goats and sheep brought to the municipal slaughterhouse of Tadjenanet, in Mila, north-eastern Algeria

A four-month survey was carried out to study the prevalence, intensity and ecological indices of Ixodidae on cattle, goats and sheep brought to the slaughterhouse in the municipality of Tadjenanet, in north-eastern Algeria. Of the 447 animals examined, 67 (14.96%) were infested by one or more ticks. A total of 262 ticks were collected and ten species grouped under two genera were inventoried: Rhipicephalus spp. (79.77%) and Hyalomma spp. (20.23%). The tick spicies collected included Hyalomma anatolicum, Hyalomma excavatum, Hyalomma impeltatum, Hyalomma lusitanicum, Hyalomma marginatum, Hyalomma scupence, Rhipicephalus (Boophilus) annulatus Rhipicephalus bursa, Rhipicephalus turanicus and Rhipicephalus sanguineus. This is the first time that Hyalomma impeltatum have been reported in Mila region. The most abundant species was Rhipicephalus bursa (41.22%). The presence of Hyalomma impeltatum in the study area was an interesting finding. The present study provides basic data on the status of the parasite in the province of Mila.

Entomological Survey for CCHF Vectors in Palghar District of Maharashtra

In view of reports of alert issued by Department of Animal Husbandry, Government of Maharashtra for Crimean Congo Hemorrhagic Fever (CCHF) in Palghar district, a visit was planned from 2-5 October 2020 to assess the situation and vector surveillance of ticks was carried out in the district. There is no past history with any report of CCHF cases in the district. However, there have been reported outbreaks of dengue and malaria during 2018 & 2019 as per IDSP data. The department of Animal Husbandry had initiated tick reduction activities in the bordering 19 villages with Gujarat using the chemical compounds (Deltamethrin, Cypermethrin). No confirmed cases of CCHF have been reported in District Palghar so far. During tick surveys in 4 villages and a dairy farm, heavy infestation of Boophilus Sp. (60-70%) mainly on cows and buffaloes.Hyalomma Sp. (30-40%) was observed mainly on goats which is the main vector responsible for CCHF. The infected ticks were found to be Boophilus microplus, Hyalomma anatolicum, Hy. marginatumand Rhipicephalus sanguineus species.Guidelines on proper dosage for the formulations of insecticide being used and spray techniques need to be followed up on the animals by the field staff and officials of Animal Husbandry department.