Gibsoni Infection Research Articles

Renal azotemia and associated haematobiochemical findings in a Labrador Retriever dog with Babesia gibsoni infection

Renal azotemia and associated haematobiochemical findings in a Labrador Retriever dog with Babesia gibsoni infection

Molecular Survey of Tick-Borne Haemoparasites of Dogs by Multiplex Polymerase Chain Reaction from Punjab, India.

Tick-transmitted parasites as Babesia gibsoni, Babesia vogeli, Ehrlichia canis, and Hepatozoon canis are major health concern for dogs. Owing to prevalence and infection severity, there is need of sensitive, specific, and affordable test for their simultaneous detection. Prevalence of B. gibsoni, B. vogeli, E. canis, and H. canis infections was assessed on 719 blood samples by microscopy and multiplex PCR assay targeting 18S rRNA (B. gibsoni & H. canis), ITS1 & 5.8S rRNA (B. vogeli) and VirB9 gene (E. canis). An internal control (canine-actin) was also included to increase the accuracy of assay and effect of associated risk factors with disease prevalence was also studied. Microscopic prevalence of B. gibsoni, B. vogeli, E. canis and H. canis was 5.0%, 0.1%, 1.4% and 1.0%, respectively, whereas with multiplex PCR assay, the corresponding values were 8.9%, 1.1%, 2.6% and 5.1% besides concurrent infections of B. gibsoni & H. canis (0.4%), B. gibsoni & E. canis (0.4%), E. canis & H. canis (0.3%) and B. gibsoni & B. vogeli (0.1%). Analytical sensitivity of developed assay was 0.1pg (B. gibsoni & H. canis), 0.01pg (B. vogeli), and 1.0pg (E. canis). A ″fair″ (B. vogeli & H. canis) to ″substantial″ (B. gibsoni & E. canis) agreement between two tests was observed with data as statistically significant. Breed, sex and location were significantly associated with B. gibsoni infection. The developed multiplex PCR assay offers a potential solution to detect these pathogens simultaneously, aiding in timely diagnosis and effective disease management in suspected dogs.

First report of occurrence of Babesia gibsoni in captive Indian wolves

Blood samples from fifteen captive Indian wolves (Canis lupus pallipes) maintained at Arignar Anna Zoological Park, Vandalur, Chennai were screened for the presence of Babesia spp., Ehrlichia canis and Trypnosoma evansi DNA by PCR. Out of 15 wolf samples, 3 samples were found positive for Babesia spp. The amplified 18S rRNA gene fragments from 3 wolves were sequenced and confirmed as Babesia gibsoni. A maximum likelihood tree was constructed using the three sequences along with other Babesia spp. sequences derived from GenBank adopting HKY nucleotide substitution model based on the Bayesian Information Criterion. The phylogenetic analysis confirmed that the three sequences were of Babesia gibsoni and highly divergent from Babesia canis, B. vogeli and B. vulpes. This might be a possible spill over event of B. gibsoni from community dogs through blood feeding dog ticks. This is the first report and molecular confirmation of B. gibsoni infection in captive Indian wolves.

Analysis of Genetic Diversity of cytb gene from Babesia gibsoni Isolates from Naturally Infected Dogs in Karnataka, India.

The study aimed to investigate genetic diversity in Babesia gibsoni, the causative agent of canine babesiosis, and to assess the presence of atovaquone-resistant isolates in naturally infected dogs. A total of 24 blood samples confirmed for B. gibsoni infection was subjected to PCR amplification and sequencing based on cytb gene. Genetic characterization of B. gibsoni as well as attempts to detect the point mutation rendering atovaquone resistance was carried out based on the analysis of nucleotide sequence of cytb gene using bioinformatics software. The findings indicated that the B. gibsoni isolates in the investigation exhibited a high nucleotide identity with the Asian genotype, ranging from 98.41 to 98.69%. Notably, none of the isolates carried cytb gene variants associated with atovaquone resistance. Phylogenetic analysis revealed clustering of most isolates with those from Japan and China, except for one isolate forming a distinct subclade. Haplotype network analysis indicated a high diversity with 22 distinct haplotypes among the B. gibsoni isolates, emphasizing the genetic variability within the studied population. In conclusion, the cytb gene exhibited remarkable conservation among the twenty-four B. gibsoni isolates studied and the study represents the first genetic diversity assessment of B. gibsoni using the cytb gene in dogs from India. These findings shed light on the genetic characteristics of B. gibsoni in the region and provide valuable insight for addressing the challenges posed by this life-threatening disease in dogs.

Evaluation of recombinant Babesia gibsoni thrombospondin-related adhesive protein (BgTRAP) for the sero-diagnosis of canine babesiosis

Canine babesiosis, caused by Babesia gibsoni is one of the most significant tick-borne illnesses across the world. Light microscopy as well as polymerase chain reaction may fail in the diagnosis of disease when the level of parasitaemia is very low during subclinical and chronic cases. The serological techniques using a recombinant protein will be useful for the accurate and sensitive surveillance of the disease, especially in chronic cases. The present study describes the evaluation of recombinant N-terminal B. gibsoni Thrombospondin-related adhesive protein (BgTRAP) based indirect ELISA for the sero-diagnosis of B. gibsoni infection in dogs. A partial N-terminal BgTRAP gene (870 bp) of B. gibsoni, was expressed in Escherichia coli using a pET32a (+) vector. The recombinant BgTRAP based indirect ELISA was compared with the PCR targeting the same gene. A sensitivity and a specificity of 84% and 73.33% were observed in the indirect ELISA. The accuracy, positive predictive value and negative predictive value were 78.18%, 72.30%, 84.60% respectively. The rBgTRAP antigen did not show any cross-reactivity with sera from dogs infected with common helminth parasites viz. Ancylostoma caninum, Dirofilaria immitis, D. repens, Spirometra spp., Toxocara canis and haemoparasites like Trypanosoma evansi, Babesia vogeli, Hepatozoon canis and Ehrlichia canis.

Clinical, Haemato-biochemical Evaluation and Comparison of Polymerase chain reaction with Conventional Blood Smear Examination for the Detection of Babesia gibsoni Infection in Dogs

The objective of the present study was to detect Babesia gibsoni infection in dogs which are presented to Veterinary Clinical Complex, Rajiv Gandhi Institute of Veterinary Education and Research, Puducherry. A total of 85 dogs with clinical signs such as inappetence, fever, enlarged lymph nodes and tick infestation were screened. The diagnostic efficacy of polymerase chain reaction (PCR) technique has been compared with blood smear examination to detect B. gibsoni infection in dogs. By blood smear and PCR examination, six (7.1%) dogs and 28 (42.4%) dogs were positive for B. gibsoni infection respectively. Clinical examination of dogs with B. gibsoni showed tick infestation, anorexia, dullness, pyrexia, pale/congested conjunctival mucus membrane, tachypnoea and swollen lymph nodes. In addition, few dogs had icterus and haemoglobinuria. Laboratory examination of blood samples of B. gibsoni dogs revealed significant reduction in haemoglobin, PCV, erythrocyte count, platelet count and serum albumin levels. Significant increase in levels of ALP, globulin, bilirubin, BUN and creatinine were recorded.

Babesia gibsoni Whole-Genome Sequencing, Assembling, Annotation, and Comparative Analysis.

The intracellular protozoan parasite Babesia gibsoni infects canine erythrocytes and causes babesiosis. The hazards to animal health have increased due to the rise of B. gibsoni infections and medication resistance. However, the lack of high-quality full-genome sequencing sets has expanded the obstacles to the development of pathogeneses, drugs, and vaccines. In this study, the whole genome of B. gibsoni was sequenced, assembled, and annotated. The genomic size of B. gibsoni was 7.94 Mbp in total. Four chromosomes with the size of 0.69 Mb, 2.10 Mb, 2.77 Mb, and 2.38 Mb, respectively, 1 apicoplast (28.4 Kb), and 1 mitochondrion (5.9 Kb) were confirmed. KEGG analysis revealed 2,641 putative proteins enriched on 316 pathways, and GO analysis showed 7,571 annotations of the nuclear genome in total. Synteny analysis showed a high correlation between B. gibsoni and B. bovis. A new divergent point of B. gibsoni occurred around 297.7 million years ago, which was earlier than that of B. bovis, B. ovata, and B. bigemina. Orthology analysis revealed 22 and 32 unique genes compared to several Babesia spp. and apicomplexan species. The metabolic pathways of B.gibsoni were characterized, pointing to a minimal size of the genome. A species-specific secretory protein SA1 and 19 homologous genes were identified. Selected specific proteins, including apetala 2 (AP2) factor, invasion-related proteins BgAMA-1 and BgRON2, and rhoptry function proteins BgWH_04g00700 were predicted, visualized, and modeled. Overall, whole-genome sequencing provided molecular-level support for the diagnosis, prevention, clinical treatment, and further research of B. gibsoni. IMPORTANCE The whole genome of B. gibsoni was first sequenced, annotated, and disclosed. The key part of genome composition, four chromosomes, was comparatively analyzed for the first time. A full-scale phylogeny evolution analysis based on the whole-genome-wide data of B. gibsoni was performed, and a new divergent point on the evolutionary path was revealed. In previous reports, molecular studies were often limited by incomplete genomic data, especially in key areas like life cycle regulation, metabolism, and host-pathogen interaction. With the whole-genome sequencing of B. gibsoni, we provide useful genetic data to encourage the exploration of new terrain and make it feasible to resolve the theoretical and practical problems of babesiosis.

Babesia gibsoni Infection in a Cat with Immune-Mediated Haemolytic Anaemia and Thrombocytopenia

Tick-borne haemoparasite Babesia gibsoni has been detected rarely in cats, in surveys of apparently healthy animals. In stored blood from a 6-year-old male-neutered domestic shorthair cat in Hong Kong, B. gibsoni DNA was detected retrospectively using PCR for Babesia spp. 18S rRNA and mitochondrial cytochrome B genes, followed by sequencing and basic local alignment search tool (BLAST) analysis. The cat presented with severe haemolytic anaemia and thrombocytopenia. The cat responded to supportive care and glucocorticoids and was clinically normal despite persistent subclinical thrombocytopenia until six months after presentation, when it succumbed to a fatal haemorrhagic episode. Necropsy revealed severe intestinal and pulmonary haemorrhage and hypocellular bone marrow with megakaryocytosis but no other causes of immune-mediated thrombocytopenia (IMTP) or immune-mediated haemolytic anaemia (IMHA). Blood stored on days 158 and 180 tested PCR negative for Babesia spp. This report demonstrates that geographic range of B. gibsoni detection in cats includes Hong Kong. The exclusion of other causes suggests that B. gibsoni might have potentially played a role in triggering immune-mediated disease in this case.

Prevalence of Babesia and Ehrlichia in owned dogs with suspected tick-borne infection in Hong Kong, and risk factors associated with Babesia gibsoni.

Babesiosis and ehrlichiosis are the most clinically significant tick-borne infections in dogs. Although epidemiological investigations of these diseases have been performed in some Asian regions, little data is available in Hong Kong, where competent vector tick species are endemic. The objectives of this study were to determine the molecular prevalence of Ehrlichia canis and Babesia species (B. canis, B. gibsoni, B. vogeli) in owned dogs suspected of tick-borne infection in Hong Kong and to identify risk factors associated with B. gibsoni infection. Electronic records from the Veterinary Diagnostic Laboratory of City University of Hong Kong were searched to identify canine blood samples submitted for molecular testing of these pathogens by real time PCR between March 2018 and May 2021. Electronic patient records from the affiliated veterinary hospital were searched to identify a subset of tested dogs to investigate the potential risk factors for B. gibsoni infection using logistic regression models. Among 1508 tested dogs for all four pathogens of interest, Babesia spp. were detected in 435 (28.8%) and E. canis in 112 (7.4%). Babesia gibsoni was detected in 408 dogs while B. vogeli was detected in 27 dogs. Babesia canis was not detected in any dog. Co-infections of different combinations of B. gibsoni, B. vogeli and E. canis were present in 25 dogs. In multivariable logistic regression, mixed breed dogs were more likely to be infected with B. gibsoni than purebreds (P=0.005), while dogs >10 years of age were less likely to be infected than younger dogs (P=0.019). Hematological abnormalities significantly associated with B. gibsoni infection included thrombocytopenia, neutropenia, or pancytopenia. Babesiosis caused by B. gibsoni is a common infection in owned dogs suspected of tick-borne infection in Hong Kong. The risk factors reported should be considered in diagnosing dogs suspected of infection with this agent. Furthermore, consideration for testing for B. gibsoni infection should be given if the results of a complete blood count show thrombocytopenia even in the absence of anemia, neutropenia or pancytopenia.

Identification of three members of the multidomain adhesion CCp family in Babesia gibsoni

Babesia gibsoni is an intraerythrocytic apicomplexan parasite transmitted by Haemaphysalis longicornis and causes canine babesiosis. Within the tick, the Babesia parasite undergoes sexual conjugation and the sporogony process of its life cycle. To control B. gibsoni infection, prompt and effective treatment of acute infections and curing chronic carriers are urgently needed. Gene disruption of Plasmodium CCps resulted in blocking the transition of sporozoites from the mosquito midgut to the salivary glands, showing that these proteins are potential targets for the development of a transmission-blocking vaccine. In this study, we described the identification and characterization of three members of the CCp family in B. gibsoni, named CCp1, CCp2, and CCp3. The B. gibsoni sexual stages were induced in vitro by exposing parasites to xanthurenic acid (XA), dithiothreitol (DTT), and tris (2-carboxyethyl) phosphine (TCEP) at serial concentrations. Among them, 100 µM XA-exposed and cultured at 27 °C without CO2B. gibsoni presented diverse morphologies, including parasites with long projections, gradually increased free merozoites, and aggregated and round forms, indicative of sexual stage induction. Then, the expression of CCp proteins of induced parasites was confirmed by real-time reverse transcription PCR, immunofluorescence, and western blot. The results showed that BgCCp genes were highly significantly increased at 24 h post-sexual stage induction (p < 0.01). The induced parasites were recognized by anti-CCp mouse antisera and anti-CCp 1, 2, and 3 antibodies weakly reacted with sexual stage proteins of expected molecular weights of 179.4, 169.8, and 140.0 KDa, respectively. Our observations on morphological changes and confirmation of sexual stage protein expression will advance elemental biological research and lay the foundation for the development of transmission-blocking vaccines against canine babesiosis.

First molecular report of Babesia gibsoni infection in pet dogs in Hunan province, subtropical China.

Canine babesiosis caused by Babesia spp. is a noteworthy tick-borne zoonotic disease of domestic dogs and wild canids. In present study, a total of 556 blood samples were randomly collected from pet dogs in eight cities of Hunan province, subtropical China. Genomic DNA was extracted and Babesia DNA was detected by amplification of partial 18S rRNA gene sequences. A total of 56 (10.1%) blood samples were tested positive for Babesia species. Sequence analysis showed that 29 dogs (5.2%) were positive for B. gibsoni, and other 27 dogs for B. vogeli (4.9%). The age and health status were considered as important risk factors for B. gibsoni and B. vogeli infections in pet dogs in this study (P<0.05). Phylogenetic analysis showed that the examined positive samples were highly clustered in the same branch with B. gibsoni and B. vogeli, respectively. This is the first molecular report of B. gibsoni infection in pet dogs in Hunan province, subtropical China. Our finding has provided a guide for the control of dog babesiosis in China and elsewhere.

Multiple point mutations in cytochrome b gene of Babesia gibsoni – A possible cause for buparvaquone resistance

The development and rise in drug-resistant Babesia gibsoni strain is a serious concern among veterinary practitioners. Of several therapeutic strategies followed, buparvaquone-azithromycin combination therapy is widely used to treat small Babesia infections in Asia. The present study focused on buparvaquone-induced mutations in B. gibsoni by analyzing its cytochrome b gene sequence. Among the 12 dogs that were administered with buparvaquone-azithromycin combination therapy, 8 of them were unresponsive to drug-resistant B. gibsoni infection. Hematological parameters were recorded before and after 10 day treatment plan and an improvement in these values was observed. Eight samples with persistent parasitemia after 10 day treatment plan was subjected to cytochrome b gene amplification and analyzed for mutations. On analysis, out of the 25 mutations only 9 were non-synonymous in nature; T15N, S48T, P152L, V167I, A217T, F258Y, M311I, S336G, A337S. Mutation P152L was seen near to Q01 (130–148) binding region and F258Y within the drug binding region Q02 (244–266).

The Etiology, Incidence, Pathogenesis, Diagnostics, and Treatment of Canine Babesiosis Caused by Babesia gibsoni Infection.

Simple SummaryBabesia gibsoni is a parasite that causes the rupture of red blood cells in dogs. Although there is no natural, tick-borne transmission of this disease in Europe, it has become more common in European countries in recent years. Dogfighting breeds are predisposed to disease and they are a potential source of infection. Given the high popularity of these dog breeds in Europe and the participation of many dogs at sports competitions and shows, it is likely that the incidence of the disease in Europe may be higher than expected. The fact that the disease is mostly manifested as asymptomatic infection and that dogs of predisposed breeds are often imported from endemic areas, or they travel due to mating or competitions around the world, also contributes to this hypothesis.Babesia gibsoni is one of the small Babesia species and the infection this pathogen causes is usually asymptomatic, which complicates the capture of potential parasite carriers. In endemic areas, especially in Asia, B. gibsoni occurs quite often due to direct transmission by way of a tick vector. Due to the absence of vectors, its occurrence is described only sporadically in Europe; but, it is increasingly occurring in predisposed, so-called fighting breeds, especially the American pit bull terrier. This review describes the etiology, incidence, clinical signs, pathogenesis, diagnostics, and treatment of B. gibsoni infection, with an emphasis on the clinical and laboratory peculiarities of the disease. As the treated dogs do not eliminate the parasite from the body—only reducing parasitemia and improving clinical signs—the treatment of B. gibsoni infection is a challenge in many cases, and its study therefore deserves great attention.

Babesia gibsoni Infection in Dogs-A European Perspective.

Simple SummaryCanine babesiosis is a tick-borne, protozoal, hemoparasitic disease caused by infection by parasites of the genus Babesia. Numerous species of Babesia exist worldwide. These protozoa are classified as either large forms (e.g., Babesia canis) or small forms (e.g., Babesia gibsoni). Reports of infections with small protozoan species are far less numerous. In most European countries where B. gibsoni has been reported, the percentage of infected dogs is around 1%. The literature review suggests that B. gibsoni should not only be considered a random and imported pathogen, but also a possible emerging parasite in Europe. The disease is much more severe than B. canis infections in most cases. Accurate molecular detection and species identification are important for selecting the correct therapy and predicting the course of the disease in dogs with babesiosis. In the future, it is expected that B. gibsoni infections may appear in other non-endemic regions in Europe, which may pose significant diagnostic and therapeutic challenges for veterinary practitioners.Canine babesiosis is a disease caused by infection with parasites of the genus Babesia. These protozoa are classified as either large (e.g., Babesia canis) or small (e.g., Babesia gibsoni). So far, only three small Babesia species of clinical importance, able to infect dogs, have been described: B. gibsoni, B. conradae, and B. vulpes. This review presents the current epidemiological situation of Babesia gibsoni infections in dogs in Europe. In most European countries where B. gibsoni has been reported, the percentage of infected dogs is around 1%. The higher prevalence of the B. gibsoni infection among American Pit Bull Terriers suggests breed susceptibility. An analysis of the available data makes it possible to conclude that B. gibsoni infections may appear in the future in other non-endemic regions of Europe, which may pose significant diagnostic and therapeutic challenges for veterinary practitioners.

Clinical characteristics of naturally Babesia gibsoni infected dogs: A study of 60 dogs.

Babesia gibsoni is increasingly recognized globally as a cause of canine tick-borne anemic disease; however, only a few clinical reports of naturally acquired infection are available. In this systematic study of dogs presenting with B. gibsoni infection, clinical and laboratory data were collected for dogs with PCR-confirmed B. gibsoni infection admitted to the National Taiwan University Veterinary Teaching Hospital (NTUVH) from January 2014 through December 2015. Of the 60 dogs recruited, 20 (33.3%) had concurrent disease and 40 (66.7%) had only B. gibsoni infection. The severity of anemia in B. gibsoni infected dogs with concurrent or without concurrent infection was not significantly different. The most commonly observed hematological abnormalities were anemia (49/60, 81.7%) and thrombocytopenia (37/60, 61.7%). Of 49 dogs, 24 (49%) had severe to very severe anemia (PCV<20%). The main biochemical abnormalities included hyperglobulinemia (28/53, 52.8%), hyperbilirubinemia (10/28, 35.7%) and elevated hepatic enzyme activity (7/48, 14.6%). In addition, 2 of the 60 the client-owned dogs and 5 of the 33 B. gibsoni-positive stray dogs were detected as having a naturally atovaquone-resistant strain, using the SimpleProbe® assay. The study results provide a useful clinical presentation of B. gibsoni infection and raise the issue of the naturally atovaquone-resistant strain currently existing in Taiwan.

Sequence and phylogenetic analysis of the thrombospondin-related adhesive protein gene of Babesia gibsoni isolates in dogs in South India.

Babesia gibsoni, the causative agent of canine piroplasmosis, is a tick-borne intraerythrocytic protozoan parasite predominantly reported in Asian countries. The present study aimed at genotypic characterization of B. gibsoni isolates prevalent in dogs in Kerala, a southern state of India. Blood samples were collected from 272 dogs in Kerala and B. gibsoni infection was detected by microscopy and polymerase chain reaction (PCR). Molecular confirmation of B. gibsoni parasites was carried out by 18S rRNA nested-PCR, followed by sequencing. Nested-PCR detected a higher percentage of dogs (40.44%) positive for B. gibsoni infection than microscopy where 15.81% dogs were detected positive for infection. Genetic characterization of B. gibsoni isolates (n=11) prevalent in dogs in the state of Kerala was carried out by PCR amplification and sequencing of the 855bp thrombospondin-related adhesive protein (TRAP) gene fragment. Phylogenetic analysis of the B. gibsoni TRAP (BgTRAP) gene revealed that B. gibsoni isolates from Kerala formed a distinct cluster with the isolates from north India and Bangladesh, away from other East Asian isolates. Nucleotide analysis of the tandem repeats of BgTRAP gene showed considerable genetic variation among Indian isolates that was shared by B. gibsoni isolates of Bangladesh but not by the isolates of East Asian countries. The results of the present study further confirmed that B. gibsoni parasites in a distinct genetic clade are endemic in dogs in India and Bangladesh. However, elaborate studies are required for better understanding of the genetic diversity of B. gibsoni.

Comparative evaluation of darbepoetin therapy in non-regenerative anaemia associated with Babesia gibsoni infection in dogs

The present study describes a comparative evaluation of haemato-therapeutic response to darbepoetin therapy in non-regenerative anaemia associated with Babesia gibsoni infection in dogs. A total of 68 dogs, presented at Teaching Veterinary Clinical Complex, Mannuthy with signs of anaemia, weakness, anorexia and pallor of mucous membranes were screened for nonregenerative anaemia. Twenty dogs with a reticulocyte count of &lt; 60,000/μL, haematocrit of &lt; 30 per cent and positive for B. gibsoni were selected for detailed study. The twenty dogs were divided into two groups of ten animals and were subjected to estimation of complete blood count and analysis of serum protein and iron status. Ten dogs brought to the hospital for vaccination or health check-up served as healthy control. Haematological examination revealed significant anaemia and thrombocytopenia in both the groups. Serum total protein, albumin and total iron binding capacity revealed no difference between diseased groups with the healthy control. Serum iron and percentage transferrin saturation were similar in Group I and II, but significantly higher than healthy control. To compare darbepoetin efficacy, Group I dogs were administered with clindamycin and doxycycline for three weeks, whereas Group II was given with darbepoetin at the dose rate of 0.5 μg/kg body weight at weekly intervals for three weeks in addition to clindamycin-doxycycline as in Group I. Dogs of both groups showed complete remission of clinical signs within 10 days of therapy. Statistical analysis revealed no significant difference in haematological parameters between the treatment groups. It can be concluded from this study that there is no added advantage of darbepoetin therapy over conventional therapies against non-regenerative anaemia associated with B. gibsoni infection in dogs.

Haemato-biochemical changes associated with Babesia gibsoni infection in dogs

The present study was conducted in dogs presented to the UVH, Kokkalai and TVCC, Mannuthy with clinical signs of inappetence, pyrexia, pale mucous membrane and lethargy suggestive of babesiosis. Forty-five dogs which were primarily screened for B. gibsoni infection through blood smear examination and then confirmed by PCR were included in the study and were subjected to a detailed physical examination, haematology and serum biochemistry analyses. Clinical outcome and variation in haematological and serum biochemical parameters were compared with values of normal dogs. Haematological analysis of affected dogs revealed normocytic normochromic anaemia, mild to moderate thrombocytopenia, inconsistent leucocyte count and differential leucocyte count revealed mild lymphocytosis and granulocytosis. Serum biochemical analysis revealed elevated alkaline phosphatase level.

Emergence of Babesia conradae infection in coyote-hunting Greyhounds in Oklahoma, USA

BackgroundBabesia species are intraerythrocytic Apicomplexan parasites that infect a wide range of vertebrate hosts. These pathogens are typically transmitted either by tick vectors or by direct blood-to-blood contact, and may cause life-threatening clinical disease, such as thrombocytopenia, hemolytic anemia and acute renal failure, in canine hosts. While Babesia vogeli and Babesia gibsoni infections have both been reported in Oklahoma, reports of Babesia conradae infections have been limited to California.MethodsFour separate kennels of coyote-hunting dogs were identified in Oklahoma after the kennels had consulted with Oklahoma State University Boren Veterinary Medical Teaching Hospital (antemortem cases) or the Oklahoma Animal Disease Diagnostic Lab (postmortem cases). Upon owner consent, every accessible dog from each of the four kennels was briefly examined for ectoparasites, particularly ticks, and whole blood samples were collected in EDTA tubes. Clinically ill dogs were examined by a practicing veterinarian, and clinical signs included anorexia, vomiting, lethargy, fever and anemia. DNA was extracted from each blood sample, and a nested PCR was performed using general apicomplexan primers for the partial 18S rRNA gene. PCR products were electrophoresed in agarose matrix, and appropriately sized amplicons were sequenced. Sequences were compared to reference 18S rRNA gene sequences available in GenBank, and samples with > 98% homology to B. conradae (GenBank: AF158702) were considered positive. Babesia conradae-positive dogs were then treated with atovaquone (13.5 mg/kg three times per day) and azithromycin (10 mg/kg once daily) for 10 days and retested at 30 and 60 days post-treatment by PCR.ResultsOf 40 dogs tested, 15 (37.5%) were positive for B. conradae with 98–99% sequence homology to B. conradae from California. All positive cases were coyote-hunting Greyhounds. Ectoparasites were not identified on any of the dogs at the time of blood collection. Treatment of clinically ill dogs with atovaquone and azithromycin resulted in complete clinical recovery in all treated dogs with negative follow-up PCR at 30 and 60 days post-treatment.ConclusionsCollectively, this study (i) documents the occurrence of B. conradae in Oklahoma, (ii) highlights this pathogen as a differential to be considered when clinical signs are present, (iii) supports the use of atovaquone and azithromycin as effective treatment in these cases and (iv) demonstrates chronic subclinical carrier dogs serving as potential reservoirs of B. conradae infection to naïve dogs.Graphical abstract

Babesia gibsoni infection in Italy: a cross sectional study of 607 blood samples belonging to dogs that needed a molecular analysis investigation (2016-2019).

Babesia gibsoni (B. gibsoni) infection was sporadically described in Europe. In Italy, in particular, it was detected by PCR analysis only in one dog with unclear source of infection, while it has never been found in molecular studies involving more subjects. The aim of this retrospective, cross-sectional study was to determine the presence and the proportion of B. gibsoni infected dogs in a convenient Italian canine population. Babesia spp PCR analysis reports of canine blood samples collected from January 2016 to December 2019 were selected from the San Marco veterinary laboratory database and evaluated. Piroplasms PCR analysis was performed on 18S rRNA gene and positive samples were sequenced. Available data about signalment, living area and season of diagnosis were evaluated and statistically analyzed. The studied canine population had a median age of 62 months (interquartile range, IQR: 72.5) and was composed of male/female purebred and mixed breed dogs. Through sequencing, B. canis (26/40, 65%), B. vogeli (7/40, 17.5%), B. gibsoni (6/40, 15%) and B. vulpes (1/40, 2.5%) infections were reported. The proportion of B. gibsoni infected dogs in the described population was of 0.99%. The infection resulted more likely in male (6/6, 100%), purebred (5/6, 83.3%) dogs with median age of 40.5 months (IQR:32.3). In one case, the clinical signs and parasitemia appeared after a dogfight with a cohabitant infected dog. No other plausible sources of infection were detected. Likely, this could be considered the first autochthonous B. gibsoni infection case documented in Italy. Our results underline the need of considering B. gibsoni not only an exotic, random and rare cause of disease for dogs in Italy, but a possible emerging pathogen to take into account during diagnostic and epidemiological investigations, in symptomatic as well as in asymptomatic patients.