Molecular Detection of<i>Theileria annulata</i>in Cattle from Different Regions of Punjab, Pakistan, by Using Recombinase Polymerase Amplification and Polymerase Chain Reaction

Hassan, Mohammad Farooque; Liu, Junlong; Sajid, Muhammad Sohail; Mahmood, Altaf; Zhao, Shuaiyang; Abbas, Qamar; Guan, Guiquan; Yin, Hong; Luo, Jianxun

doi:10.1645/17-173

Cited by 11 publications

(9 citation statements)

References 22 publications

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“…Concurrently, research should also be focused on unexplored areas of population genomics of TTBPs using high-throughput techniques such as NGS and microfluidics. As discussed earlier, the current focus of research is on the control of TTBDis of large ruminants in this country [72,119,[160][161][162], however, in a predominantly mixed-species farming system where small and large ruminants are reared together, sheep and goats could serve as alternative hosts for TTBPs of large ruminants [163,164]. Therefore, for the effective control of TTBDis of livestock in Pakistan, it is imperative to design future studies including common livestock species that are kept in proximity.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Systematic Review of Ticks and Tick-Borne Pathogens of Small Ruminants in Pakistan

et al. 2020

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Ticks and tick-borne diseases (TTBDis) are a major constraint to the health and production of small ruminants in Pakistan. Despite being the subject of intermittent studies over the past few decades, comprehensive information on the epidemiology and control of TTBDis is lacking. Herein, we have systematically reviewed the current knowledge on TTBDis of small ruminants in Pakistan. Critical appraisal of the selected 71 articles published between 1947 to 2020 revealed that morphological examination had been the most widely used method for the identification of TTBDis in Pakistan. Tick fauna comprise at least 40 species, mainly belonging to Haemaphysalis, Hyalomma and Rhipicephalus. The prevalence of ticks is the highest in summer (June–September) and it is also higher in goats than sheep. Anaplasma, Babesia and Theileria spp. are the major tick-borne pathogens (TBPs), and their prevalence is usually higher in sheep than goats. Spatio-temporal distribution, genetic diversity and control of ticks and TBPs of small ruminants as well as the competence of tick vectors for various TBPs remain to be explored. Therefore, coordinated and focused investigations are required to fill knowledge gaps in these areas to maximise the health, production and welfare of small ruminants and minimise economic losses associated with TTBDis in Pakistan.

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Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Systematic Review of Ticks and Tick-Borne Pathogens of Small Ruminants in Pakistan

et al. 2020

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“…Furthermore, previous studies were confined to the surrounding areas of major metropolitan cities without considering diverse climatic conditions and bovine production systems in Pakistan which are important factors in the design of epidemiological studies [7]. Recently, several studies have utilized molecular methods for the detection of pathogens in bovines [30, 31] and their ticks [18]. However, these studies only targeted the main TBPs pathogens (e.g.…”

Section: Introductionmentioning

confidence: 99%

Bovine ticks harbour a diverse array of microorganisms in Pakistan

et al. 2020

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BackgroundTicks and tick-borne pathogens (TTBP) are a major constraint to livestock production in Pakistan; despite a high prevalence of TTBPs, knowledge on the capacity of Pakistani ticks to carry pathogens and endosymbionts is limited. Furthermore, mixed infections with multiple microorganisms further complicate and limit the detection potential of traditional diagnostic methods. The present study investigated the tick-borne microorganisms in bovine ticks in Pakistan, employing a high-throughput microfluidic real-time PCR based technique.MethodsTicks were collected from clinically healthy cattle (n = 116) and water buffaloes (n = 88) from 30 villages across six districts located in five agro-ecological zones (AEZs) of Pakistan from September to November 2017. The microfluidic real-time PCR was used to test the genomic DNA of individual ticks for the presence of 27 bacterial and eight parasitic microorganisms. Phylogenetic methods were used to assess the genetic relationship of DNA sequences determined herein.ResultsPCR detected DNA of at least one microorganism in each of 221 ticks tested (94.4%, 221/234). DNA-based detection inferred that single pathogens/endosymbionts were the most common (43.4%, 96/221) followed by double (38.9%, 86/221), triple (14.5%, 32/221), quadruple (2.3%, 5/221) and quintuple (0.9%, 2/221) mixed infections. Piroplasms (Babesia/Theileria spp.) were the most prevalent (31.6%, 74/234), followed by Ehrlichia spp. (20%, 47/234) and Anaplasma marginale (7.7%, 18/234). Anaplasma phagocytophilum, A. ovis, A. centrale, Babesia ovis, Borrelia spp., Rickettsia spp., R. massiliae, Bartonella spp. and Hepatozoon spp. were also detected. Endosymbionts such as Francisella-like (91.5%, 214/234) and Coxiella-like (1.3%, 3/234) organisms were also detected in ticks. The highest diversity of microorganisms was detected in Hyalomma anatolicum ticks (test-positive for 14/14 microorganisms), followed by Rhipicephalus microplus (4/14), Hy. hussaini (3/14) and Rh. annulatus (2/14). Ticks collected from cattle carried significantly more frequently piroplasms (41.2%, 54/131; P < 0.05) than those from buffaloes (19.4%, 20/103). However, the overall prevalence of microorganisms did not vary significantly among ticks from the two host species as well as across different AEZs.ConclusionsTo our knowledge, this is the first study to investigate a wide range of tick-borne microorganisms in bovine ticks using a high-throughput diagnostic method from different AEZs in Pakistan. These findings will aid in establishing the distribution patterns and the control of tick-borne pathogens of bovines in Pakistan.

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“…Gebrekidan et al [ 20 ] reported the presence of T. orientalis in Pakistan for the first time from imported and native animals, reporting no T. orientalis in native animals by conventional PCR and low prevalence (6%) in native cattle as compared with imported animals (25.4%) using multiplexed tandem PCR. Recently, Hassan et al [ 21 , 22 ] determined the presence of Theileria spp. by using recombinase polymerase amplification.…”

Section: Introductionmentioning

confidence: 99%

Molecular survey of piroplasm species from selected areas of China and Pakistan

Hassan¹,

Liu

Rashid

et al. 2018

Parasites Vectors

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BackgroundPiroplasmosis is an important animal disease that is a major constraint to the development of the livestock industry, often resulting in significant economic losses. Therefore, there is an urgent need to further understand the etiology of this and other tick-borne infections.MethodsBlood samples were collected from tick-infested animals from the Chakwal, Jhang, and Faisalabad districts of Punjab, Pakistan and from peri-urban areas around Hohhot, Inner Mongolia, China to investigate the presence of Babesia and Theileria species. In total, 450 blood samples were collected with FTA cards from cattle of the study areas of Pakistan; the genomic (g)DNA of one hundred and twenty samples from cattle in Inner Mongolia were provided by the Lanzhou Veterinary Research Institute, China. Following the extraction of gDNA, the 18S rRNA gene (V4 hypervariable region) of piroplasms was amplified in all samples using semi-nested PCR. Positively identified samples were sequenced for the identification of Theileria and Babesia species. The partial full-length sequence of 18S rDNA was amplified for species confirmation of Theileria-positive samples, whereas the RAP-1c gene was amplified for Babesia bigemina-positive samples.ResultsSemi-nested PCR results revealed that 144 (25.26%) samples were positive for piroplasms. Theileria annulata was the most prevalent species (115/144; 20.17%), followed by Theileria orientalis (16/144; 2.80%). Among Babesia, the only species recorded was Babesia bigemina (13/144; 2.28%).ConclusionThe present study reveals new data on the prevalence of piroplasm species in bovine populations of selected areas of China and Pakistan and their phylogenetic relationships. It is also the first detailed report of T. orientalis from native animals in Pakistan.

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Molecular Detection ofTheileria annulatain Cattle from Different Regions of Punjab, Pakistan, by Using Recombinase Polymerase Amplification and Polymerase Chain Reaction

Cited by 11 publications

References 22 publications

Systematic Review of Ticks and Tick-Borne Pathogens of Small Ruminants in Pakistan

Systematic Review of Ticks and Tick-Borne Pathogens of Small Ruminants in Pakistan

Bovine ticks harbour a diverse array of microorganisms in Pakistan

Molecular survey of piroplasm species from selected areas of China and Pakistan

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