Serological and molecular diagnostic surveys combined with examining hematological profiles suggests increased levels of infection and hematological response of cattle to babesiosis infections compared to native buffaloes in Egypt

Particle and Fibre Toxicology

Carlos E. Suarez²  Marta G. Silva⁴  Dalia M. Mabrouk³  Salwa M. Habeeb¹  Seham H.M. Hendawy¹  Soad M. Nasr¹  Omnia M. Kandil¹  Mona S. Mahmoud¹ 

[1] Parasitology and Animal Diseases Department, National Research Center, 33 Bohouth St., Giza 12622, Dokki, Egypt;Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Pullman, WA, USA;Cell biology Department, National Research Center, 33 Bohouth St., Giza 12622, Dokki, Egypt;Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA

关键词: Egypt;    Hemogram;    Semi-nested PCR;    Nested PCR;    Competitive ELISA;    B. bovis;    B. bigemina;   

Others  :  1224202
DOI  :  10.1186/s13071-015-0928-9

 received in 2015-01-23, accepted in 2015-06-02,  发布年份 2015

【 摘 要 】

Background

Babesiosis threatens the development of the cattle and buffaloes industries in Egypt and improved control is needed. The main objectives of this study are surveying the presence of bovine babesiosis in distinct selected bovine and buffalo populations in Egypt using novel molecular and previously validated serological methods, while also comparing the occurrence of hematological alterations among Babesia infected cattle and buffalos.

Methods

A total of 253 and 81 blood samples from apparently healthy cattle and buffaloes, respectively, were randomly collected from diverse locations in Egypt. All samples were tested for Babesia bovis and B. bigemina infection using blood film examination, competitive ELISA (cELISA) and PCR. Novel semi-nested and nested PCR assays for the detection of B. bovis and B. bigemina respectively, were developed and used to analyze DNA extracted from bovine and buffalo samples. Hematological profiles were studied using a hematological analyzer.

Results

Blood films examination revealed 13.8 % and 7.4 % Babesia infection rates in cattle and buffaloes, respectively. However, in cattle, the cELISA detected 32.8 %, 21.3 % and 10.7 % infection rates with B. bigemina, B. bovis and mixed infection, respectively. In addition, cELISA identified 22.2 %, 22.2 % and 6.2 % infection rates with B. bigemina, B. bovis and mixed infection, respectively in buffaloes. The semi-nested PCR assay showed that 15 % of the tested samples were positive for B. bovis in cattle, but just 3 % in buffaloes. Infections with B. bigemina were also found in cattle (32.4 %), but not in buffaloes upon nested PCR analysis. Sequencing analysis confirmed the identity of the PCR amplicons and showed that Egyptian genotypes of B. bigemina and B. bovis highly resemble sequences previously deposited in GenBank. Hemograms performed on the sampled animals revealed macrocytic hypochromic anemia associated with reduced platelet counts in infected cattle with babesiosis. In addition, marked increases in total leukocyte and granulocytic counts and decreases in lymphocytic counts were found in infected cattle. In contrast, no such hematological anomalies were found in presumably Babesia-infected buffaloes.

Conclusions

Frequent occurrence of babesiosis among apparently healthy bovines in Egypt, suggests the need for appropriately designed prevalence studies in this country. Infected bovine, but not buffalo, populations often present hematological disorders compatible with intravascular hemolysis and thrombocytopenia.

【 授权许可】

   
2015 Mahmoud et al.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

【 参考文献 】

• [1]Zintl A, Mulcahy G, Skerrett HE, Taylor SM, Gray JS. Babesia divergens, a bovine blood parasite of veterinary and zoonotic importance. Clin Microbiol Rev. 2003; 16(4):622-636.
• [2]Mosqueda J, Olvera-Ramirez A, Aguilar-Tipacamu G, Canto GJ. Current advances in detection and treatment of babesiosis. Curr Med Chem. 2012; 19(10):1504-1518.
• [3]Spickler ARaJA R. Emerging and Exotic Diseases of Animal, 3rd ed. CFSPH Iowa State University, Ames, Iowa; 2008.
• [4]Lozano DFA. Modelling of Parasitic Diseases with Vector OF Transmission: Toxoplasmosis and Babesiosis Bovine. Universidad Politecnica de Valenci, Departamento de Matematica Aplicada; 2011.
• [5]Adham FK, Abd-el-Samie EM, Gabre RM, El-Hussein H. Detection of tick blood parasites in Egypt using PCR assay I--Babesia bovis and Babesia bigemina. Parasitol Res. 2009; 105(3):721-730.
• [6]Abdel-Shafy S, Mona S. Mahmoud: Morphological and ultrastructural studies on Babesia bigemina local strain in haemolymph and salivary glands of infected Boophilus annulatus females. J Egypt Vet Med Assoc. 2002; 62(6):247-256.
• [7]Messick JB. Hemotrophic mycoplasmas (hemoplasmas): a review and new insights into pathogenic potential. Veterinary clinical pathology/American Society for Veterinary Clinical Pathology. 2004; 33(1):2-13.
• [8]Goes TS, Goes VS, Ribeiro MF, Gontijo CM. Bovine babesiosis: anti-erythrocyte antibodies purification from the sera of naturally infected cattle. Vet Immunol Immunopathol. 2007; 116(3-4):215-218.
• [9]LP. Mechanisms of Thrombocytopenia in Tick-Borne Diseases. Internet J Infect Dis. 2002;2(2). https://ispub.com/IJID/2/2/3023
• [10]Mahmmod YS. Molecular detection of natural Babesia bovis infectioon from clinically infected and apparently healthy water buffaloes (Bubalis bubalis) and crossbred cattle. J Buffalo Sci. 2012; 1:55-60.
• [11]Elsify A, Sivakumar T, Nayel M, Salama A, Elkhtam A, Rizk M et al.. An epidemiological survey of bovine Babesia and Theileria parasites in cattle, buffaloes, and sheep in Egypt. Parasitol Int. 2014; 64(1):79-85.
• [12]Bashir HA. Modeling of development time for hydro electric generators using factor and multiple regression analyses. J Project Manage. 2008; 26:457-464.
• [13]Bose R, Jorgensen WK, Dalgliesh RJ, Friedhoff KT, de Vos AJ. Current state and future trends in the diagnosis of babesiosis. Vet Parasitol. 1995; 57(1-3):61-74.
• [14]Salih DA, El Hussein AM, Seitzer U, Ahmed JS. Epidemiological studies on tick-borne diseases of cattle in Central Equatoria State, Southern Sudan. Parasitol Res. 2007; 101(4):1035-1044.
• [15]Jacobson LS. The South African form of severe and complicated canine babesiosis: clinical advances 1994-2004. Vet Parasitol. 2006; 138(1-2):126-139.
• [16]Araujo FR, Madruga CR, Leal CR, Schenk MA, Kessler RH, Marques AP et al.. Comparison between enzyme-linked immunosorbent assay, indirect fluorescent antibody and rapid conglutination tests in detecting antibodies against Babesia bovis. Vet Parasitol. 1998; 74(2-4):101-108.
• [17]Goff WL, Molloy JB, Johnson WC, Suarez CE, Pino I, Rhalem A et al.. Validation of a competitive enzyme-linked immunosorbent assay for detection of antibodies against Babesia bovis. Clin Vaccine Immunol. 2006; 13(11):1212-1216.
• [18]Goff WL, McElwain TF, Suarez CE, Johnson WC, Brown WC, Norimine J et al.. Competitive enzyme-linked immunosorbent assay based on a rhoptry-associated protein 1 epitope specifically identifies Babesia bovis-infected cattle. Clin Diagn Lab Immunol. 2003; 10(1):38-43.
• [19]Goff WL, Johnson WC, Molloy JB, Jorgensen WK, Waldron SJ, Figueroa JV et al.. Validation of a competitive enzyme-linked immunosorbent assay for detection of Babesia bigemina antibodies in cattle. Clin Vaccine Immunol. 2008; 15(9):1316-1321.
• [20]Sharma A, Das Singla L, Tuli A, Kaur P, Batth BK, Javed M et al.. Molecular prevalence of Babesia bigemina and Trypanosoma evansi in dairy animals from Punjab, India, by duplex PCR: a step forward to the detection and management of concurrent latent infections. BioMed rsesearch international. 2013; 2013:893862.
• [21]Fahrimal Y, Goff WL, Jasmer DP. Detection of Babesia bovis carrier cattle by using polymerase chain reaction amplification of parasite DNA. J Clin Microbiol. 1992; 30(6):1374-1379.
• [22]Figueroa JV, Chieves LP, Johnson GS, Buening GM. Detection of Babesia bigemina-infected carriers by polymerase chain reaction amplification. J Clin Microbiol. 1992; 30(10):2576-2582.
• [23]Hunfeld KP, Hildebrandt A, Gray JS. Babesiosis: recent insights into an ancient disease. Int J Parasitol. 2008; 38(11):1219-1237.
• [24]Tang YW, Procop GW, Persing DH. Molecular diagnostics of infectious diseases. Clin Chem. 1997; 43(11):2021-2038.
• [25]Suarez CE, Palmer GH, Florin-Christensen M, Hines SA, Hotzel I, McElwain TF. Organization, transcription, and expression of rhoptry associated protein genes in the Babesia bigemina rap-1 locus. Mol Biochem Parasitol. 2003; 127(2):101-112.
• [26]Suarez CE, Laughery JM, Bastos RG, Johnson WC, Norimine J, Asenzo G et al.. A novel neutralization sensitive and subdominant RAP-1-related antigen (RRA) is expressed by Babesia bovis merozoites. Parasitology. 2011; 138(7):809-818.
• [27]Suarez CE, Laughery JM, Schneider DA, Sondgeroth KS, McElwain TF. Acute and persistent infection by a transfected Mo7 strain of Babesia bovis. Mol Biochem Parasitol. 2012; 185(1):52-57.
• [28]Figueroa JV, Chieves LP, Johnson GS, Buening GM. Multiplex polymerase chain reaction based assay for the detection of Babesia bigemina, Babesia bovis and Anaplasma marginale DNA in bovine blood. Vet Parasitol. 1993; 50(1-2):69-81.
• [29]Snedecor GW. CWG: Statistical Methods. 7th ed. The State University Press American, Iowa; 1989.
• [30]El-Ghaysh A. Studies on Babesia species infecting cattle and water Buffaloes in Egypt. Faculty of Veterinary Medicine, Cairo University, Egypt; 1993.
• [31]El-Fayomy AO, Ghoneim AM, Abu-Samak OA, Khidr AA. Contribution of Babesia to the Illness of Cows in Port Said Governorate. Egypt Glob Vet. 2013; 11:118-122.
• [32]Adel EM. Studies on some blood parasites infecting farm animals in Gharbia Governorate. Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt; 2007.
• [33]Abd-El Gawad M. Some studies on Babesia species in cattle in Beni-Suef governorate. Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt; 1993.
• [34]Mazyad SA, Khalaf SA. Studies on theileria and babesia infecting live and slaughtered animals in Al Arish and El Hasanah, North Sinai Governorate, Egypt. J Egypt Soc Parasitol. 2002; 32(2):601-610.
• [35]Dominguez M, Echaide I, de Echaide ST, Wilkowsky S, Zabal O, Mosqueda JJ et al.. Validation and field evaluation of a competitive enzyme-linked immunosorbent assay for diagnosis of Babesia bovis infections in Argentina. Clin Vaccine Immunol. 2012; 19(6):924-928.
• [36]Ibrahim HMAMP, Mohammed-Gebaa K, Sheira SK, Ihab SY, Hashemc S, Caob MA et al.. Molecular and serological prevalence of Babesia bigemina and Babesia bovis in cattle and water buffalos under small-scale dairy farming in Beheira and Faiyum Provinces. Egypt Vet Parasitol. 2013; 198:187-192.
• [37]Bock R, Jackson L, de Vos A, Jorgensen W. Babesiosis of cattle. Parasitology. 2004; 129 Suppl:S247-S269.
• [38]Ibrahim A, EL Behairy AM, Mahran KA, Awad WS. Clinical and laboratory diagnosis of piroplasmids in naturally infected cattle in Egypt. J Egypt Vet Med Assoc. 2009; 69:191-203.
• [39]Silva MG, Henriques G, Sanchez C, Marques PX, Suarez CE, Oliva A. First survey for Babesia bovis and Babesia bigemina infection in cattle from Central and Southern regions of Portugal using serological and DNA detection methods. Vet Parasitol. 2009; 166(1-2):66-72.
• [40]Terkawi MA, Thekisoe OM, Katsande C, Latif AA, Mans BJ, Matthee O et al.. Serological survey of Babesia bovis and Babesia bigemina in cattle in South Africa. Vet Parasitol. 2011; 182(2-4):337-342.
• [41]World Organization for Animal Health [OIE]. Bovine babesiosis. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.04.02_BOVINE_BABESIOSIS.pdf. (Online) 2008.
• [42]Wright IG, Goodger BV, Leatch G, Aylward JH, Rode-Bramanis K, Waltisbuhl DJ. Protection of Babesia bigemina-immune animals against subsequent challenge with virulent Babesia bovis. Infect Immun. 1987; 55(2):364-368.
• [43]Rania YE. Some studies on diagnosis on babesiosis. Faculty of Veterinary Medicine, Benha University, Benha, Egypt; 2009.
• [44]Chaudhry ZI, Suleman M, Younus M, Aslim A. Molecular Detection of Babesia bigemina and Babesia bovis in Crossbred Carrier Cattle through PCR. Pakistan J Zool. 2010; 42:201-204.
• [45]Yu L, Terkawi MA, Cruz-Flores MJ, Claveria FG, Aboge GO, Yamagishi J et al.. Epidemiological survey of Babesia bovis and Babesia bigemina infections of cattle in Philippines. The Journal of veterinary medical science/the Japanese Society of Veterinary Science. 2013; 75(7):995-998.
• [46]L’Hostis M, Seegers H. Tick-borne parasitic diseases in cattle: current knowledge and prospective risk analysis related to the ongoing evolution in French cattle farming systems. Vet Res. 2002; 33(5):599-611.
• [47]Coles GC. The future of veterinary parasitology. Vet Parasitol. 2001; 98(1-3):31-39.
• [48]Zulfiqar S, Shahnawaz S, Ali M, Bhutta AM, Iqbal S, Hayat S et al.. Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab. Asian Pacific journal of tropical biomedicine. 2012; 2(2):104-108.
• [49]Pandy NaM S. Haematological and Biochemical response to haemolytic anaemia of clinical Babesiosis in cattle and therapy. Ind Vet J. 1987; 64:882-886.
• [50]Murase TaM Y. Increased erythophagocytosis activity of macrophages in dogs with Babesia gibsoni infection. Nippon Juigaku Zasshi. 1990; 52:321-327.
• [51]Court RA, Jackson LA, Lee RP. Elevated anti-parasitic activity in peripheral blood monocytes and neutrophils of cattle infected with Babesia bovis. Int J Parasitol. 2001; 31(1):29-37.
• [52]Taboada JaL R. Babesiosis. In: Infectious diseases of the dog and Cat. 3rd ed. Greene CE, editor. Saunders Elsevier, St Louis; 2006: p.722-736.
• [53]Riond BMM, Braun U, Deplazes P, Joerger K, Thoma R, Lutz H et al.. Concurrent infections with vector-borne pathogens associated with fatal anaemia in cattle: Haematology and blood chemistry. Comp Clin Pathol. 2008; 17(3):171-177.
• [54]Musaji A, Cormont F, Thirion G, Cambiaso CL, Coutelier JP. Exacerbation of autoantibody-mediated thrombocytopenic purpura by infection with mouse viruses. Blood. 2004; 104(7):2102-2106.
• [55]Mahmmod Y. Natural Babesia bovis Infection in Water Buffaloes (Bubalus bubalis) and Crossbred Cattle under Field Conditions in Egypt: a Preliminary Study. J Arthropod-Borne Dis. 2014; 8(1):1-9.