Sunanda CHULLIYIL, Lavinia WAHLANG, Aravindakshan THAZHATHUVEETIL, Bindu LAKSHMANAN, Naicy THOMAS, Jain JOSE, Anu BOSEWELL

SYBR green-based real-time PCR detection of canine Babesia spp. in ixodid ticks infesting dogs in Kerala, South India

Ticks, being hematophagous ectoparasites, are notorious vectors that transmit an array of pathogenic viruses, bacteria, andprotozoa while simultaneously exhibiting a zoonotic potential by transmitting pathogens that affect the health of owners in contact.The distribution of ixodid ticks of the genera Rhipicephalus and Haemaphysalis spp. in tropical climate of the state contributes to manyserious tick-borne parasitic and rickettsial infections in domestic and wild canines. In south India, canine babesiosis is one of the mostprevalent vector-borne parasitic diseases. Detection of parasites in tick vectors has significant epidemiological implications. The presentstudy was designed to identify the presence of the most common vector-borne parasites of dogs in ixodid ticks using sensitive detectionprotocols. SYBR green-based real-time protocols detected Babesia vogeli in Rhipicephalus sanguineus sensu lato, while B. gibsoni wasdetected in R. sanguineus (s.l.) and Haemaphysalis bispinosa. The present study points out the need to reinvestigate the vectorial capacityof ticks in different geographical regions.

PDF

___

1. Otranto D, Testini G, Dantas-Torres F, Latrofa MS, de Paiva Diniz et al. Diagnosis of canine vector-borne diseases in young dogs: a longitudinal study. Journal of Clinical Microbiology 2010; 48(9): 3316-24.
2. Rani APA, Irwin PJ, Coleman GT, Gatne M, Traub RJ. A survey of canine tick-borne diseases in India. Parasites and Vectors 2011; 4: 141.
3. Kundu K, Kumar S, Mandal MM, Ram H, Garg R et al. PCR based identification of Babesia canis vogeli in clinically affected dogs. Journal of Veterinary Parasitology 2012; 26: 167-169.
4. Augustine S, Sabu L, Lakshmanan B. Molecular identification of Babesia spp. in naturally infected dogs of Kerala, South India. Journal of Parasitic Diseases 2017; 2: 459-462.
5. Jain J, Lakshmanan B, Nagaraj HV, Praveena JE, Syamala K et al. Detection of Babesia canis vogeli, Babesia gibsoni and Ehrlichia canis by multiplex PCR in naturally infected dogs in South India. Veterinarski Arhiv 2018; 88: 215-224.
6. Jose KJ, Lakshmanan B, Wahlang L, Syamala K, Aravindakshan TV. Molecular evidence of haemoparasites in ixodid ticks of dogs-first report in India. Veterinary Parasitology Regional Studies and Reports 2018; 13: 177-179.
7. Solano-Gallego L, Baneth G. Babesiosis in dogs and cats- Expanding parasitological and clinical spectra. Veterinary Parasitology 2011; 181: 48-60.
8. Hamel D, Silaghi C, Zapadynska S, Kurdin A, Pfster K. Vectorborne pathogens in ticks and EDTA-blood samples collected from client-owned dogs Kiev Ukraine. Ticks and Tick Borne Diseases 2013; 4: 152-155.
9. Harrus S, Warner T. Diagnosis of canine monocytotropic ehrlichiosis (Ehrlichia canis): an overview. Veterinary Journal 2010; 187: 292-296.
10. Paiva-Cavalcanti M, Regis-da-Silva CG, Gomes YM. Comparison of real-time PCR and conventional PCR for detection of Leishmania (Leishmania) infantum infection: a mini-review. Journal of Venomous Animals and Toxins including Tropical Diseases 2010; 16: 537-542.
11. Costa- Junior LM, Zahler- Rinder M, Ribeiro MFB, Rembeck K, Rabelo EML et al. Use of a Real time PCR for detecting subspecies of Babesia canis. Veterinary Parasitology 2012; 188: 160-163.
12. Cardoso L, Oliveira AC, Granada S, Nachum-Biala Y, Gilad et al. Molecular investigation of tick-borne pathogens in dogs from Luanda, Angola. Parasites and Vectors 2016; 9: 252.
13. Sen SK, Fletcher TB. Veterinary Entomology and Acarology in India. New Delhi, India: Indian Council of Agricultural Research; 1962.
14. Rodriguez I, Fraga J, Noda AA, Mayet M, Duarte Y et al. An alternative rapid method for the extraction of nucleic acid from ixodid ticks by potassium acetate procedure. Brazilian Archives of Biology and Technology 2014; 57: 542-547.
15. M’ghirbi Y, Bouattour A. Detection and molecular characterization of Babesia canis vogeli from naturally infected dogs and Rhipicephalus sanguineus ticks in Tunisia.Veterinary Parasitology 2008; 152: 1-7.
16. Trotta M, Nicetto M, Fogliazza A, Montarsi F, Caldin M et al. Detection of Leishmania infantum, Babesia canis, and rickettsiae in ticks removed from dogs living in Italy. Ticks and Tick-borne Diseases 2012; 3: 294-297.
17. Tsuji N, Battsetseg B, Boldbaatar D, Miyoshi T, Xuan X et al. Babesial vector tick defensin against Babesia sp. parasites. Infection and Immunity 2007; 75: 3633-3640.
18. Taboada J, Lobetti R. Babesiosis Infectious Diseases of the Dog and Cat. Missouri, USA: Saunders Elsevier; 2006.
19. Smythe LD, Smith IL, Smith GA, Dhont MF, Symonds ML et al. A quantitative PCR assay for pathogenic Leptospira spp. Journal of BioMed Central Infectious Diseases 2002; 2: 1-7.
20. Zemtsova GE, Montgomery M, Levin ML. Relative sensitivity of conventional and real-time PCR assays for detection of SFG Rickettsia in blood and tissue samples from laboratory animals. PloS one 2015; 10: 1-7.
21. Burggraf S, Reischl U, Malik N, Bollwein M, Naumann L et al. Comparison of an internally controlled, large-volume Light Cycler assay for detection of Mycobacterium tuberculosis in clinical samples with the COBAS AMPLICOR assay. Journal of Clinical Microbiology 2005; 43: 1564-1569.
22. Yera H, Delhaes L, Filisetti D, Ancelle T, Thulliez P et al. Multicentre comparison of five DNA extraction methods for Toxoplasma gondii from amniotic fluid Journal of Clinical Microbiology 2009; 47: 3881-3886.
23. Bastien P, Procop GW, Reischl U. Quantitative real-time PCR is not more sensitive than “conventional” PCR. Journal of Clinical Microbiology 2008; 46: 1897-1900.