Assessment of exposure to piroplasms in sheep grazing in communal mountain pastures by using a multiplex DNA bead-based suspension array

Particle and Fibre Toxicology

Ana Hurtado¹  Ramón A Juste¹  Ana L García-Pérez¹  Jesús F Barandika¹  Amaia Ros-García¹ 

[1] Department of Animal Health, NEIKER - Instituto Vasco de Investigación y Desarrollo Agrario, Berreaga 1, Derio, Bizkaia 48160, Spain

关键词: Piroplasmosis;    Tick-borne pathogens;    RLB;    Sheep;    Suspension microarray;    Luminex;    Theileria;    Babesia;   

Others  :  824487
DOI  :  10.1186/1756-3305-6-277

 received in 2013-07-02, accepted in 2013-09-17,  发布年份 2013

【 摘 要 】

Background

Piroplasms are tick-borne hemoprotozoans with a major impact on extensive management systems. Detection of sub-clinical low-level carriers, which can act as source of infection for vector ticks, is key to protect livestock trade and facilitate preventive control programs. The purpose of this study was to develop a method for the detection of ovine piroplasms and to use it in a field study aimed at investigating piroplasms infection in semi-extensive production systems in the Basque Country (northern Spain).

Methods

A DNA bead-based suspension array using the Luminex® xMAP technology that included a generic Theileria-Babesia control probe, 6 species-specific probes, and an internal control probe was developed to detect and identify piroplasms that infect sheep. To monitor piroplasm infection in clinically healthy sheep from 4 flocks that share communal mountain pastures, blood samples were collected during 2 grazing seasons.

Results

Piroplasms were detected in 48% (214/446) of blood samples, nearly half of them (49.1%, 105/214) as mixed infections. Five different piroplasms were identified: Theileria sp. OT3 in 34.8% of the samples, Theileria ovis in 20.9%, and at lower prevalences Babesia motasi (12.3%), Theileria luwenshuni/OT1 (10.5%) and Babesia ovis (6.3%). Despite differences among flocks associated to differences in management, an increasing trend in the incidence of piroplasm infection with increasing age of animals after increased tick exposure was observed. This increment could be attributed to continued re-infection associated with re-exposure to ticks at grazing. Ticks were collected from animals (4 species) and vegetation (8 species), and associations between tick abundance seasonality and risk of infection with the different piroplasms were established.

Conclusion

The multiplex Luminex® xMAP procedure is a rapid and high throughput technique that provided highly specific and sensitive identification of single and mixed piroplasm infections in blood of sheep carriers. This study confirmed a situation of endemic stability for piroplasm infection in the region, where infection is present in the absence of clinical signs, and mountain grazing allows for sufficient inoculation rates to maintain such situation.

【 授权许可】

   
2013 Ros-García et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Gabiña D, Arrese F, Arranz J, Beltran de Heredia I: Average milk yields and environmental effects of Latxa sheep. J Dairy Sci 1993, 76:1191-1198.
• [2]Nagore D, García-Sanmartín J, García-Pérez AL, Juste RA, Hurtado A: Identification, genetic diversity and prevalence of Theileria and Babesia species in a sheep population from Northern Spain. Int J Parasitol 2004, 34:1059-1067.
• [3]Preston PM: Theilerioses. In Encyclopedia of arthropod-transmitted infections of man and domesticated animals. Edited by Service M.W. Wallingford, UK: CABI Publishing; 2001:487-502.
• [4]Uilenberg G: Babesia-A historical overview. Vet Parasitol 2006, 138:3-10.
• [5]Yin H, Luo J, Schnittger L, Lu B, Beyer D, Ma M, Guan G, Bai Q, Lu C, Ahmed J: Phylogenetic analysis of Theileria species transmitted by Haemaphysalis qinghaiensis. Parasitol Res 2004, 92:36-42.
• [6]Yin H, Luo J, Guan G, Lu B, Ma M, Zhang Q, Lu W, Lu C, Ahmed J: Experiments on transmission of an unidentified Theileria sp. to small ruminants with Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum. Vet Parasitol 2002, 108:21-30.
• [7]Uilenberg G: Babesiosis. In Encyclopedia of arthropod-transmitted infections of man and domesticated animals. Edited by Service M.W. Wallingford, UK: CABI Publishing; 2001:53-60.
• [8]Sonenshine DE: Biology of ticks, Volume II. New York: Oxford University Press; 1993.
• [9]Chauvin A, Moreau E, Bonnet S, Plantard O, Malandrin L: Babesia and its hosts: adaptation to long-lasting interactions as a way to achieve efficient transmission. Vet Res 2009, 40:37.
• [10]Yeruham I, Hadani A, Galker F: Some epizootiological and clinical aspects of ovine babesiosis caused by Babesia ovis–a review. Vet Parasitol 1998, 74:153-163.
• [11]Liu AH, Yin H, Guan GQ, Schnittger L, Liu ZJ, Ma ML, Dang ZS, Liu JL, Ren QY, Bai Q, Ahmed JS, Luo JX: At least two genetically distinct large Babesia species infective to sheep and goats in China. Vet Parasitol 2007, 147:246-251.
• [12]Schnittger L, Yin H, Qi B, Gubbels JM, Beyer D, Niemann S, Jongejan E, Ahmed JS: Simultaneous detection and differentiation of Theileria and Babesia parasites infecting small ruminants by reverse line blotting. Parasitol Res 2004, 92:189-196.
• [13]Ros-García A, Juste RA, Hurtado A: A highly sensitive DNA bead-based suspension array for the detection and species identification of bovine piroplasms. Int J Parasitol 2012, 42:207-214.
• [14]Gil-Collado J, Guillén-Llera JL, Zapatero-Ramos LM: Claves para la identificación de los Ixodoidea españoles (adultos). Rev Iber Parasitol 1979, 39:107-111.
• [15]Manilla G: Fauna D’Italia: Acari, Ixodida. Bologna: Edizioni Calderini; 1998.
• [16]Georges K, Loria GR, Riili S, Greco A, Caracappa S, Jongejan F, Sparagano O: Detection of haemoparasites in cattle by reverse line blot hybridisation with a note on the distribution of ticks in Sicily. Vet Parasitol 2001, 99:273-286.
• [17]Juste RA, Garrido JM, Geijo M, Elguezabal N, Aduriz G, Atxaerandio R, Sevilla I: Comparison of blood polymerase chain reaction and enzyme-linked immunosorbent assay for detection of Mycobacterium avium subsp. paratuberculosis infection in cattle and sheep. J Vet Diagn Invest 2005, 17:354-359.
• [18]Gubbels JM, de Vos AP, van der Weide M, Viseras J, Schouls LM, de Vries E, Jongejan F: Simultaneous detection of bovine Theileria and Babesia species by reverse line blot hybridization. J Clin Microbiol 1999, 37:1782-1789.
• [19]Hoorfar J, Malorny B, Abdulmawjood A, Cook N, Wagner M, Fach P: Practical considerations in design of internal amplification controls for diagnostic PCR assays. J Clin Microbiol 2004, 42:1863-1868.
• [20]Barandika JF, Hurtado A, Juste RA, García-Pérez AL: Seasonal dynamics of Ixodes ricinus in a 3-year period in northern Spain: first survey on the presence of tick-borne encephalitis virus. Vector Borne Zoonotic Dis 2010, 10:1027-1035.
• [21]Barandika JF, Olmeda SA, Casado-Nistal MA, Hurtado A, Juste RA, Valcarcel F, Anda P, García-Pérez AL: Differences in questing tick species distribution between Atlantic and continental climate regions in Spain. J Med Entomol 2011, 48:13-19.
• [22]García-Sanmartín J, Barandika JF, García-Pérez AL, Juste RA, Hurtado A: Distribution and molecular detection of Theileria and Babesia in questing ticks from Northern Spain. Med Vet Entomol 2008, 22:318-325.
• [23]Barandika JF, Berriatua E, Barral M, Juste RA, Anda P, García-Pérez AL: Risk factors associated with ixodid tick species distributions in the Basque region in Spain. Med Vet Entomol 2006, 20:177-188.
• [24]Yeruham I, Hadani A, Galker F, Rosen S: A study of an enzootic focus of sheep babesiosis (Babesia ovis, Babes, 1892). Vet Parasitol 1995, 60:349-354.
• [25]Norval RAI, Perry BD, Young AS: The Epidemiology of Theileriosis in Africa. London, UK: Academic Press; 1992.
• [26]Dalgliesh RJ: Babesiosis . In Immunology and molecular biology of parasitic infections. 3rd edition. Edited by Warren KS. Boston: Blackwell Scientific Publications; 1993.
• [27]Wikel SK: Tick modulation of host immunity: an important factor in pathogen transmission. Int J Parasitol 1999, 29:851-859.