A new PCR assay for the detection and differentiation of Babesia canis and Babesia vogeli

Background and Aim: Canine babesiosis, a tick-borne parasitic disease, is caused by the hemoprotozoa, Babesia vogeli, and Babesia gibsoni. Infection with these parasites, which is endemic globally, leads to life-threatening immunosuppression in dogs. The merozoites invade the red blood cells (RBCs) of infected dogs. Ehrlichia canis, an intracellular bacterium that infects monocytes, is transmitted by the same tick species (Rhipicephalus sanguineus) during blood consumption and coinfection with B. vogeli and E. canis has been reported. Although the hematology and biochemistry of canine babesiosis have been studied, more studies are needed to develop a better understanding of the hematobiochemical and molecular profiles associated with cases of single infection and coinfection of canine babesiosis in Thailand. This study aimed to investigate the hematological, biochemical, and molecular profiles of B. vogeli infection and E. canis coinfection. Materials and Methods: The study included 33 B. vogeli–positive blood samples and 11 E. canis–coinfected blood samples. To exclude coinfection with Hepatozoon canis and Anaplasma platys, only dogs with B. vogeli infection and B. vogeli–E. canis coinfection were included in the study. A multiplex polymerase chain reaction (PCR) assay was conducted to detect B. vogeli, E. canis, and H. canis, and a conventional PCR assay was conducted for the detection of A. platys. Besides, the PCR assay and sequencing, comprehensive data analysis was conducted, including a microscopic blood parasite examination and hematological and biochemical data analysis. Results: The comparison of the hematobiochemical data between the B. vogeli–positive and E. canis coinfection groups identified that there were statistically significant differences in the RBC parameters, including RBC count, hemoglobin concentration, hematocrit, and RBC distribution width (p=0.001). Neither B. vogeli infection nor coinfection with E. canis was associated with the sex, breed, recorded clinical signs, geographic origin of the dog and also B. vogeli 18S rRNA gene sequencing results. Conclusion: Coinfection with E. canis increased the severity of babesiosis. The pathogenic mechanisms underlying this infection, such as destruction of RBCs, require further investigation. This study may enhance diagnosis, treatment, and prevention of canine babesiosis.

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Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular approach

Parasitology Research ◽

10.1007/s00436-011-2683-y ◽

2011 ◽

Vol 110 (5) ◽

pp. 1659-1664 ◽

Cited By ~ 22

Author(s):

Mariana Ionita ◽

Ioan Liviu Mitrea ◽

Kurt Pfister ◽

Dietmar Hamel ◽

Catalin Marius Buzatu ◽

...

Keyword(s):

Babesia Canis ◽

Molecular Approach ◽

Canine Babesiosis ◽

Babesia Vogeli

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Assessment of primers designed for the subspecies-specific discrimination among Babesia canis canis, Babesia canis vogeli and Babesia canis rossi by PCR assay

Veterinary Parasitology ◽

10.1016/j.vetpar.2007.12.013 ◽

2008 ◽

Vol 152 (1-2) ◽

pp. 16-20 ◽

Cited By ~ 22

Author(s):

Sabrina Castilho Duarte ◽

Guido Fontgalland Coelho Linhares ◽

Tatiana Nunes Romanowsky ◽

Osvaldo José da Silveira Neto ◽

Ligia Miranda Ferreira Borges

Keyword(s):

Babesia Canis ◽

Pcr Assay ◽

Babesia Canis Canis

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Detection and Molecular Characterization of Babesia canis vogeli and Theileria annulata in Free-Ranging Dogs and Ticks from Shahriar County, Tehran Province, Iran

Iranian Journal of Parasitology ◽

10.18502/ijpa.v15i3.4196 ◽

2020 ◽

Author(s):

Gholamreza HABIBI ◽

Alireza IMANI ◽

Asghar AFSHARI ◽

Soghra BOZORGI

Keyword(s):

Molecular Techniques ◽

Babesia Canis ◽

Pcr Assay ◽

Blood Samples ◽

Tick Borne Disease ◽

Dog Blood ◽

Vector Borne ◽

Bovine Theileriosis ◽

Free Ranging

Background: We aimed to detect and characterize vector-borne parasites of Babesia and Theileria in dog and ticks by PCR assay. Canine babesiosis is a significant tick-borne disease caused by different Babesia species. As the infection has not been reported in Shahriar region Tehran, Iran, molecular techniques allowed us to identify tick-borne parasites in asymptomatic dogs. Methods: The number of 40 dog peripheral blood samples and 27 skin attached ticks were analyzed by molecular PCR assay. The specific primers were used for detecting Babesia canis, B. gibsoni and T. annulata. Results: B. c. vogeli was detected in 10 dog blood samples (25%). Additionally, T. annulata infection was identified in 13 dog blood samples (32.5%) and 18 isolated tick DNAs (66.7%). The results of PCR were confirmed by 18S rRNA and Tams1 gene sequence analyzing and have been registered in GenBank under following accession numbers for B. c. vogeli (MH793502) and T. annulata (MK105284). Conclusion: The verification of T. annulata infection in free-ranging dogs and ticks shows dogs might be considered as important natural carriers/reservoirs for T. annulata in enzootic region for bovine theileriosis. The obtained data may be useful for veterinary practitioners and dog owners to aware of Babesia and Theileria infection in dog and tick to establish the effective preventive measures.

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Genetic Detection and Phylogenetic Relationship of Babesia Species Infecting Domestic Dogs from Select Regions in Kenya

10.21203/rs.3.rs-95605/v3 ◽

2021 ◽

Author(s):

Ismail Thoya Ngoka ◽

Kevin Mbogo ◽

Martina Kyallo ◽

David Obiero ◽

Roger Pelle

Keyword(s):

Phylogenetic Relationship ◽

Vaccine Development ◽

Severe Disease ◽

Current Control ◽

Control Measures ◽

Babesia Canis ◽

Canine Babesiosis ◽

Study Objective ◽

Babesia Vogeli ◽

Babesia Rossi

Abstract The genus Babesia has more than 100 species that are transmitted by ticks with some being zoonotic. They can infect humans, livestock, and wildlife. Although canine babesiosis occurs locally, published studies on the species involved are limited. Babesia parasites cause severe disease in dogs which can be fatal. Drawbacks of the current control methods necessitate vaccine development. The study objective was to identify the Babesia species infecting dogs from three Kenyan counties; Nairobi, Mombasa, Nakuru and determine their phylogenetic relationship. This will enable improved control and rule out zoonotic potential. The study period was October 2018 to November 2019.The study design was descriptive and sampling opportunistic. One hundred and forty-three dogs were sampled. From whole blood, total DNA was extracted using the TanBead extractor followed by PCR amplification targeting Babesia 18S rRNA. Positive samples were purified and sequenced using the Sanger Dideoxy method.CLC Genomics Workbench, GenBank™ and BLASTn™ on NCBI were used for sequence processing and analysis. Geneious prime™ was used for multiple sequence alignment and phylogenetic analysis. The overall prevalence of Babesia canis was 9.0% (95% CI: 4.37–13.81). Two out of 13 positive samples (2/13) were identified as Babesia canis vogeli, with a prevalence of 1.4% (95% CI: 1.38–14.2, n = 143) while 11/13 were identified as Babesia canis rossi, with a prevalence of 7.69% (95% CI: 3.3–12, n = 143). The Babesia rossi sequences identified were closely related to sequences from black-backed jackals, while the Babesia vogeli ones were related to sequences from a pet cat in China. Babesia rossi which causes severe canine babesiosis was identified in 84.6% of the positive samples, immediate and aggressive clinical intervention is necessary. The possible sylvatic cycle of Babesia rossi and low levels of infections by Babesia vogeli should inform pertinent control measures.

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Molecular Investigation of Canine Babesiosis in and Around Bhubaneswar, India

10.21203/rs.3.rs-237874/v1 ◽

2021 ◽

Author(s):

Charchika Panda ◽

Manaswini Dehuri ◽

Mitra Ranjan Panda ◽

Lakshman Kumar Sahoo ◽

Bijayendranath Mohanty

Keyword(s):

Hepatozoon Canis ◽

Gene Fragment ◽

Canine Babesiosis ◽

Pcr Assay ◽

Blast Analysis ◽

Babesia Vogeli ◽

Molecular Investigation ◽

Pcr Products ◽

Microscopic Diagnosis ◽

Genetic Sequence Analysis

Abstract Canine babesiosis, a hemolytic protozoan disease represents an important veterinary problem caused primarily by large and small forms of piroplasms of Babesia spp. A molecular-based survey on the overall occurrence of natural Babesia infection in stray(n=98 ) and pet dogs(n=100) from Bhubaneswar and nearby areas using PCR technique targeting 18s RNA gene fragment along with genetic sequence analysis was carried out. A total of 38 (pet:22, stray:16) samples (19.19%) were found positive for babesiosis based on the amplification of 450 bp amplicon region of the gene while 4 samples (0.02%) showed co-infection with Hepatozoon canis. The sequenced PCR products were submitted to NCBI ,and on BLAST analysis the isolates with accession no KT246303, KT246306, KT246307 showing similarity with Babesia vogeli, while KT246305 was identical to B.gibsoni isolates and KT246304 was identical to Hepatozoon canis. This is the first report on the molecular diagnosis of canine babesiosis in the state. PCR assay was found to be more precise over microscopic diagnosis, the use of more specific and sensitive tests along with more samples could aid in a better understanding of the epidemiology of canine babesiosis in this region.

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Development and validation of a multiplex, real-time PCR assay for Babesia rossi and Babesia vogeli

Ticks and Tick-borne Diseases ◽

10.1016/j.ttbdis.2018.12.004 ◽

2019 ◽

Vol 10 (2) ◽

pp. 421-432 ◽

Cited By ~ 2

Author(s):

Milana Troskie ◽

Lourens de Villiers ◽

Andrew Leisewitz ◽

Marinda C. Oosthuizen ◽

Melvyn Quan

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Assay ◽

Babesia Vogeli ◽

Babesia Rossi ◽

Development And Validation

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Detection and molecular characterization of piroplasms species from naturally infected dogs in southeast Brazil

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612012000200012 ◽

2012 ◽

Vol 21 (2) ◽

pp. 137-142 ◽

Cited By ~ 17

Author(s):

Tatiana Didonet Lemos ◽

Aloysio de Mello Figueiredo Cerqueira ◽

Helena Keiko Toma ◽

Adrianna Vieira da Silva ◽

Rafael Gomes Bartolomeu Corrêa ◽

...

Keyword(s):

Rio De Janeiro ◽

Fragment Length Polymorphism ◽

Restriction Enzymes ◽

18S Rrna Gene ◽

Rrna Gene ◽

Sequencing Analysis ◽

Pcr Assay ◽

Babesia Vogeli ◽

Babesia Spp

Rangelia vitalii is a protozoon described from dogs in the south and southeast regions of Brazil. It is phylogenetically related to Babesia spp. that infects dogs, but data on this enigmatic parasite is still limited. The aim of this work was to detect piroplasm species in dogs in the state of Rio de Janeiro, Brazil, by 18S rRNA gene-based PCR assay, restriction fragment length polymorphism (RFLP) and sequence analyses. Of 103 dogs examined, seven (6.8%) were positive for Babesia spp. by PCR. The amplified products were digested by restriction enzymes to differentiate the Babesia species, and one sample was identified as Babesia vogeli. The pattern observed for the other six amplification products did not match with pattern described for large Babesia infecting dogs. Sequencing analysis confirmed these six samples as R. vitalii, with high homologies (99-100%) with a sequence from south Brazil. This study confirms the presence of Babesia vogeli and Rangelia vitalii circulate in domestic dogs in Teresópolis, Rio de Janeiro, Brazil.

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