Occurrence of Babesia Species and Co-Infection with Hepatozoon canis in Symptomatic Dogs and in Their Ticks in Eastern Romania

Although the distribution of Babesia spp. and Hepatozoon canis is well known in Romania, there is still a marked lack of information in many places of the country. This study aimed to investigate the occurrence of these haemoparasites in symptomatic dogs and in their ticks in Iasi, eastern Romania. Ninety owned dogs were subjected to clinical examination at the Faculty of Veterinary Medicine of Iasi and all detectable ticks (58 ticks from 15 dogs) were collected. Additionally, 124 ticks collected from the coat of other dogs (no. = 23) were included. Three Babesia species were found in dogs: Babesia canis (94.4%), Babesia vogeli (3.3%), and Babesia rossi (2.2%). All the dogs resulted negative for H. canis. The ticks were identified as follows: Ixodes ricinus (64%), Dermacentor reticulatus (33%), and Rhipicephalus sanguineus group (3%). B. canis (Minimum Infection Rate; MIR = 81%), B. vogeli (MIR = 3%), and Babesia microti-like piroplasm (MIR = 1%) were found in ticks. Moreover, 15 ticks were positive for H. canis, 6 were co-infected with B. canis, and 1 with B. microti-like piroplasm. This is the first molecular identification of B. rossi in two symptomatic dogs from Romania, although further studies are needed to investigate the vector competence of other ticks from Europe.

Morphological, molecular and MALDI-TOF MS identification of ticks and tick-associated pathogens in Vietnam

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been reported as a promising and reliable tool for arthropod identification, including the identification of alcohol-preserved ticks based on extracted leg protein spectra. In this study, the legs of 361 ticks collected in Vietnam, including 251 Rhiphicephalus sanguineus s.l, 99 Rhipicephalus (Boophilus) microplus, two Amblyomma varanensis, seven Dermacentor auratus, one Dermacentor compactus, and one Amblyomma sp. were submitted for MALDI-TOF MS analyses. Spectral analysis showed intra-species reproducibility and inter-species specificity and the spectra of 329 (91%) specimens were of excellent quality. The blind test of 310 spectra remaining after updating the database with 19 spectra revealed that all were correctly identified with log score values (LSV) ranging from 1.7 to 2.396 with a mean of 1.982 ± 0.142 and a median of 1.971. The DNA of several microorganisms including Anaplasma platys, Anaplasma phagocytophilum, Anaplasma marginale, Ehrlichia rustica, Babesia vogeli, Theileria sinensis, and Theileria orientalis were detected in 25 ticks. Co-infection by A. phagocytophilum and T. sinensis was found in one Rh. (B) microplus.

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

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.

Seroprevalence and prevalence of Babesia vogeli in clinically healthy dogs and their ticks in Costa Rica

Canine babesiosis is a disease caused by a parasite of the genus Babesia which destroys red blood cells. Previous studies have shown the presence of Babesia vogeli in rural areas in Costa Rica using molecular techniques. The objective of the present study was to determine the seroprevalence and prevalence of B. vogeli in clinically healthy dogs and their ticks at the national level, both within and outside the Central Valley. Blood samples and ticks from 482 dogs were collected between June 2011 and May 2014, and analyzed by immunofluorescence assay (IFA) and real-time polymerase chain reaction (qPCR); two protocols of endpoint PCR and sequencing were used to confirm qPCR-positive samples. Seroprevalence of canine babesiosis of 5.3% (24/453) was determined at the national level, specifically 2.0% (5/253) within and 9.5% (19/200) outside the Central Valley, respectively. Real-time PCR determined a global prevalence of B. vogeli of 31.3% (125/400): 21.4% (47/220) within the Central Valley and 43.3% (78/180) outside the Central Valley. The endpoint PCR amplified only 10 of the 125 blood samples identified as positive in qPCR. One sample amplified by endpoint PCR was sequenced and identified as B. vogeli. Twelve canines were identified with past infections, seven canines with active infection, and 111 canines with early infection. Two species of ticks were found with B. vogeli: Rhipicephalus sanguineus sensu lato (n = 40) and Amblyomma ovale (n = 1). The prevalence of canine babesiosis at the national level, both within and outside the Central Valley, is reported here for the first time, determining the presence of the piroplasmid throughout the country, with a higher circulation of the agent outside the Central Valley. Only one species, B. vogeli, was detected in the blood of dogs and their ticks. Therefore, veterinarians should consider using qPCR to determine the presence of the parasite in blood donors and before starting treatment of vector-borne disease in dogs.

Detection of Borrelia burgdorferi s.l., Anaplasma phagocytophilum and Babesia spp. in Dermacentor reticulatus ticks found within the city of Białystok, Poland—first data

Pathogens carried by ticks pose a threat to both human and animal health across the world. Typically associated with rural landscapes, ticks appear to adapt well to life in urban recreational areas. Although Dermacentor reticulatus is commonly found across Europe, data on the prevalence of pathogens in this tick species, in an urban environment, are very limited. PCR was used to examine 368 D. reticulatus individuals collected in the Zwierzyniecki Forest Nature Reserve in Białystok, Poland. In total, 10.3% of ticks were infected, with Babesia spp. (9.2%), Anaplasma phagocytophilum (0.8%) and Borrelia burgdorferi sensu lato (0.3%). Rickettsia spp., Bartonella spp., and Coxiella burnetii were not detected. Sequence analysis for Babesia-positive samples identified 79.4% of them as Babesia canis, 8.8% as Babesia microti, 5.9% as Babesia spp., 2.9% as Babesia venatorum, and 2.9% as Babesia vogeli. Results obtained in this study indicate that D. reticulatus ticks found within the urban premises of the study area are infected with at least three pathogens and therefore are an important factor in public health risk for tick-borne diseases.

Babesia vogeli in dogs from Rio Branco, South-west Amazonia, Brazil

This is the first report of Babesia vogeli molecular detection in dogs from the state of Acre, northern Brazil. This study aimed to perform the molecular detection of Babesia vogeli in dogs in the municipality of Rio Branco, Acre. Blood samples were collected from 47 dogs presenting with clinical signs comparable to hemoparasitosis. These were dogs which were attended in veterinary clinics from Rio Branco municipality, Acre. Physical examinations, packed cell volume (PCV) determination, platelet number estimation, hemoparasite investigation in the blood (collected from the pinna and peripheral blood), and polymerase chain reaction (PCR) for piroplasm based on the 18S rRNA gene, were performed. One dog (1/47, 2.1%; CI 95%: 0.1-11.3%) tested positive to Babesia vogeli in the polymerase chain reaction (PCR) assay for piroplasms and the resulting sequence showed 100% identity with Babesia vogeli isolates deposited in GenBank®. Co-infection with Ehrlichia spp. was also observed by direct examination (via blood smear). The clinical and hematological alterations observed in the positive animal were anorexia, dehydration, white mucous membranes, anemia and thrombocytopenia.

Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific

The Asia-Pacific hosts a large diversity of canine vector-borne pathogens (VBPs) with some of the most common and most pathogenic, generating significant mortality as well as a spectrum of health impacts on local dog populations. The VBPs Anaplasma platys, Babesia gibsoni, Babesia vogeli, Ehrlichia canis, Hepatozoon canis and haemotropic Mycoplasma spp. are all endemic throughout the region, with many exhibiting shifting geographical distributions that warrant urgent attention. Moreover, many of these species cause similar clinical signs when parasitising canine hosts, whilst knowledge of the exact pathogen is critical to ensure treatment is effective. This is complicated by frequent coinfection that can exacerbate pathology. Here, we describe the development, optimisation and validation of two novel quadruplex Taq-Man based real-time PCRs (qPCRs) for the specific and sensitive detection of the aforementioned VBPs. To ensure accurate evaluation of diagnostic performance, results of our qPCRs were evaluated on field samples from Thai dogs and compared with both conventional PCR (cPCR) results and next-generation sequencing (NGS) metabarcoding. Our qPCRs were found to be more sensitive at detecting canine VBP than cPCR and generated results similar to those achieved by NGS. These qPCRs will provide a valuable high-throughput diagnostic tool available to epidemiologists, researchers and clinicians for the diagnosis of key canine VBPs in the Asia-Pacific and further afield.

Prevalence, genetic, and biochemical evaluation of immune response of police dogs infected with Babesia vogeli

Background and Aim: Babesia species are tick-borne protozoan parasites of apicomplexan type which infect the erythrocytes of dogs it ranges from subclinical to severe cases, depending on different factors such as immune status, age, and presence of other co-infections with the Babesia species. Hence, this study aimed to identify the protozoan parasites infecting police dogs of different breeds, ages, and both sexes in Egypt. Concerning molecular detection of Babesia vogeli using conventional polymerase chain reaction sequencing and phylogenetic analysis, followed by the assessment of immunological and biochemical status of infected dogs. Materials and Methods: The blood of 242 police K9 dogs was collected. The age, breed, sex, and health status with clinical signs of dogs were recorded. Hematological, biochemical, and oxidative stress analyses of the blood were performed together with gene expression analysis using two genes (gamma interferon [IFN-γ] and tumor necrosis factor-alpha [TNF- α]). The identification of the causative agent was performed using molecular analysis of the 18S ribosomal RNA (rRNA). The 18S rRNA region of canine Babesia spp. was successfully amplified, and sequencing data were deposited in GenBank (accession number: MT565474.1), which resembled those of B. vogeli. Results: The results of blood samples screening revealed that of the 242 blood samples, 62 were positive for B. vogeli infection. The infection rate in male dogs was higher than that in female dogs. The police dogs were classified into the following three groups of dogs: (1st group) healthy, (2nd infected with B. vogeli, and mixed infection of B. vogeli and Ehrlichia canis). The oxidative stress biomarkers levels in B. vogeli infected dogs were greater than that of healthy dogs. Likewise, IFN-γ and TNF-α level in B. vogeli infected dogs were elevated in infected dogs. Conclusion: Our findings demonstrated that B. vogeli had completely adverse effects on the health condition of the police dogs that may lead to death in some dogs.

Molecular Investigation of Canine Babesiosis in and Around Bhubaneswar, India

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.