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      A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan) Translated title: Étude sur la détermination des facteurs de risque associés à la babésiose et à la présence de Babesia sp., par amplification par PCR, chez les petits ruminants du Penjab du Sud (Pakistan)

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          Abstract

          Babesiosis is a parasitic infection due to the multiplication of tick borne parasite, Babesia sp., in erythrocytes of host, which includes a wide variety of vertebrates including small ruminants causing decreased livestock output and hence economic losses. The objective of the present study was to establish a PCR based method for the detection of Babesia sp. in small ruminant population in Southern Punjab and to determine the risk factors involve in the spread of babesiosis. A total of 107 blood samples were collected from 40 sheep and 67 goats in seven districts of Southern Punjab from randomly selected herds. Data on the characteristics of the animals and the herd were collected through questionnaires. 36 blood samples (34% of total) produced the DNA fragment specific for 18S rRNA gene of Babesia sp., by PCR amplification, of which 20 were sheep and 16 were goats. Samples from all seven district contained Babesia positive samples and prevalence varied between 18 to 68%. It was observed that male animals (P = 0.009) and young animals under one year of age (P = 0.01) were more prone to the parasite. It was observed that herds consist of more than 15 animals (P = 0.007), composed of mixed species of small ruminants (P = 0.022), associated with dogs (P = 0.003) and dogs having ticks on their bodies (P = 0.011) were among the major risk factors for the spread of babesiosis in small ruminants.

          Translated abstract

          La babésiose est une infection due à la multiplication du parasite Babesia sp., transmis par des tiques, dans les érythrocytes de nombreux hôtes vertébrés dont les petits ruminants, chez qui il est responsable d’importantes pertes de productivité. Les objectifs de l’étude étaient de mettre au point une méthode de détection de Babesia sp., reposant sur la PCR, dans des populations de petits ruminants du Penjab du Sud, et de determiner les facteurs de risque impliqués dans la diffusion de la babésiose. 107 échantillons de sang ont été prélevés de façon aléatoire chez 40 ovins et 67 caprins dans des troupeaux de sept districts du Penjab du Sud. Les données concernant les caractéristiques des animaux et des troupeaux ont été collectées à l’aide d’un questionnaire. 36 échantillons de sang (34 % du total) ont révélé la présence du gène spécifique de l’ARNr 18S de Babesia sp., par amplification par PCR, chez 20 ovins et 16 caprins. Des échantillons positifs pour Babesia ont été trouvés dans les sept districts, avec une fréquence variant de 18 à 68 %. Les animaux de sexe mâle (P = 0,009) et les jeunes de moins d’un an (P = 0,01) étaient les plus touchés par le parasite. Les principaux facteurs de risque de diffusion de la babésiose chez les petits ruminants sont : les troupeaux de plus de 15 têtes (P = 0,007), composés d’un mélange de plusieurs espèces (P = 0,022), associés à la presence de chiens (P = 0,003) et à des chiens porteurs de tiques (P = 0,011).

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          Most cited references 7

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          Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe. Part I. Epizootiological aspects.

          Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced. Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following: Piroplasmid infections: Approximately from 50 to 70% of wild or domestic mammals (symptomatic) were infected. Piroplasmids detected in ruminants were:COW: B. bovis, T. annulata and Theileria sp. (type C). Sheep and goat: B. ovis. Piroplasmids present in canids were: Babesia canis vogeli, Babesia canis canis, Theileria annae and B. equi. The only piroplasmid found in asymptomatic dogs was B. equi. Piroplasmids found in horse were: B. equi and B. canis canis.H. canis infections in canids: H. canis was absent of domestic dog samples, whereas all foxes studied were infected by this protozoa. Genetic analysis showed that most of piroplasmid and Hepatozoon isolates from southern Europe matched unambigously with previously described species, as demonstrated by the high level sequence identity between them, usually between 99 and 100%. Minor differences, usually detected in hypervariable regions of 18s rRNA gene are probably due to strain variations or rare genetic polymorphisms. A possible exception was B. bovis, which shows a relatively lower degree of homology (94%) with regard to other B. bovis isolates from several countries. The same is true for B. ovis, that showed a 94% identity with regard to Babesia sp. from South African cow and a 92% with rapport to B. bovis from Portugal.
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            The piroplasms: life cycle and sexual stages.

             H Mehlhorn,  E Shein (1983)
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              Identification, genetic diversity and prevalence of Theileria and Babesia species in a sheep population from Northern Spain.

              The genetic diversity and prevalence of virtually all Theileria and Babesia species in a sheep population were studied using a specifically designed reverse line blot macroarray. The amplified hypervariable V4 region of the 18S rRNA gene was hybridised against generic and species-specific probes. In a first screening (Study I), 320 apparently healthy animals corresponding to 32 flocks located in the Basque Country (Northern Spain) were analysed. The survey demonstrated a high prevalence of subclinical infections (64.7%). Three Theileria genotypes were identified, sharing 96.7-97.0% similarity between their 18S rRNA gene sequences: Theileria ovis, Theileria sp. OT1 (99.6% similarity with the recently described pathogenic piroplasm Theileria sp. China 1), and Theileria sp. OT3. Two Babesia species sharing 91.5% similarity were also detected: Babesia ovis and Babesia motasi. The complete 18S rRNA gene sequences of these and other piroplasm species were phylogenetically analysed. Prevalence of piroplasms was also investigated in a second group of 80 sheep from 16 flocks reared in mountain areas that had been heavily exposed to ticks and had suffered a recent abortion episode (Study II). The screening revealed a significantly higher (P < 0.05) prevalence (78.7%) of piroplasm infections compared to Study I. Although the prevalence rates for some piroplasm species were significantly related to abortion (e.g. Theileria sp. OT3), decreases in the red cell parameters were not significant. The widespread distribution of Theileria spp. in the studied sheep population suggests that the parasites involved are of relatively low pathogenicity, in contrast to what has been reported for Theileria sp. China 1 in other countries.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite : journal de la Société Française de Parasitologie
                EDP Sciences
                1252-607X
                1776-1042
                August 2011
                15 August 2011
                : 18
                : 3 ( publisher-idID: parasite/2011/03 )
                : 229-234
                Affiliations
                [1 ] Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University Multan Pakistan
                [2 ] Institute of Biotechnology, Bahauddin Zakariya University Multan Pakistan
                [3 ] Department of Parasitology, Faculty of Veterinary Medicine, University of Firat 23119 Elazig Turkey
                [4 ] Faculty of Veterinary Sciences, Bahauddin Zakariya University Multan Pakistan
                Author notes
                [* ]Correspondence: Furhan Iqbal, Ph.D., Department of Zoology, Institute of Pure and Applied Biology, Bahauddin Zakariya University Multan 60800, Pakistan. Tel.: 92 61 9210053 – Fax : 92 61 9210098. E-mail: furhan.iqbal@ 123456bzu.edu.pk
                Article
                parasite2011183p229 10.1051/parasite/2011183229
                10.1051/parasite/2011183229
                3671477
                21894263
                © PRINCEPS Editions, Paris, 2011

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 1, Tables: 3, Equations: 0, References: 22, Pages: 6
                Categories
                Original Contribution

                goats, sheep, pcr amplification, ovin, caprin, pcr, amplification, babesia sp

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