Candidatus Mycoplasma Haematoparvum Research Articles

Molecular detection and identification of hemotropic Mycoplasma species in dogs and their ectoparasites in Iran

Hemotropic Mycoplasma species are vector-borne bacteria that attach and grow on the surface of erythrocytes in various mammals, yet reports of canine hemoplasmosis in Iran are scarce. The aim of this study was molecular detection and identification of hemoplasmas in the blood of dogs (n = 370) from five provinces of Iran and ectoparasites infesting them including Ctenocephalides canis and Pulex irritans fleas, Rhipicephalus sanguineus sensu lato ticks, Heterodoxus spiniger lice and Hippobosca longipennis keds. Hemotropic Mycoplasma spp. pathogens were detected using genus-specific conventional PCRs, and subsequently identified using species-specific PCRs for Mycoplasma haemocanis (Mhc), and Candidatus Mycoplasma haematoparvum (CMhp). Sanger sequencing was then performed to confirm the species. Correlation of infection and risk factors (geographical area, keeping condition, body condition, sex, age, ectoparasite infestation) were analyzed. In total, 210 dogs (56.7%) were tested PCR-positive for hemotropic Mycoplasma spp. Species-specific PCR and sequencing revealed infection with Mhc in 17.8%, with CMhp in 7.02% and co-infection in 31.9% of dogs. Flea infestation, poor body condition, and being older than 3-years-old correlated with hemoplasmosis. In ectoparasites, DNA of hemoplasmas were detected only in fleas i.e. Mhc in P. irritans, CMhp in P. irritans and C. canis, and co-infection in C. canis. To our knowledge, this is the first large-scale molecular epidemiology study of canine hemoplasmosis in Iran. Considering the high prevalence of canine hemoplasmosis all over the country including potentially zoonotic CMhp, effective ectoparasite control strategies, regular examination of dogs, successful chemoprophylaxis and public awareness strategies are advocated.

Transmission of haemotropic mycoplasma in the absence of arthropod vectors within a closed population of dogs on ectoparasiticides

Dog-infecting haemotropic mycoplasmas (haemoplasmas), such as Mycoplasmahaemocanis and Candidatus Mycoplasma haematoparvum are common blood-borne pathogens of canines that can potentially inflict a substantial burden of disease, particularly in immunosuppressed individuals. Nonetheless, the transmission of these pathogens remains debated as more evidence emerges that they may not be transmitted by vectors, but instead use alternative methods such as aggressive interactions and vertical transmission. Here, we treated forty dogs with two different topically-acting ectoparasiticide products able to prevent vector-borne pathogen infections during an 8-month community trial in Cambodia. A total absence of ectoparasites were observed at all time points, and no new infections caused by pathogens confirmed as being vectorially-transmitted were detected, i.e., Babesiavogeli, Ehrlichiacanis, Anaplasmaplatys, and Hepatozooncanis. Conversely, the number of haemoplasma infections in dogs on both ectoparasiticides rose significantly, with an incidence of 26 infections per 100 dogs at risk per year, providing strong evidence of non-vectorial transmission. Over the study period, dog aggression and fighting were frequently observed, highlighting a different potential mode of transmission. This study presents the first robust evidence that canine haemoplasmas may be transmitted without arthropod vectors drawing attention to the need for new methods to prevent their transmission.

Prevalence of Bartonella spp., haemotropic Mycoplasma spp. and others vector-borne pathogens in private-owned dogs and cats, Egypt

Vector-borne pathogens have been increasingly investigated for their impact on dog and cat health and their zoonotic potential. The aim of this study was to investigate the prevalence estimates of selected vector-borne pathogens in client-owned pets from the Giza and Cairo governorates, Egypt. Out of 200 dogs and 100 cats, 94 (47%) and 23 (23%) were positive for at least one of the tested pathogens (P<0.0001). In particular, 84 (42%) dogs and 3 (3%) cats tested PCR-positive for Bartonella spp. (P<0.0001). A significantly higher prevalence of Bartonella spp. was detected in dogs from the rural areas of the Giza governorate (60/77, 79.2%, P<0.0001) compared to those from Cairo governorate. Bartonella henselae was the dominant species infecting dogs (81/200, 40.5%) followed by Candidatus Bartonella merieuxii (3/200, 1.5%), while B. henselae (2/100, 2%) and B. clarridgeiae were rare in cats. Haemoplasma DNA was detected in 17% (34/200) of dogs and 20% (20/100) of cats with increased risk in dogs from Giza rural areas (21/77, 27.27%, P=0.002) and from both dogs (16/63, 25.40%, P=0.03) and cats (7/14, 50%, P<0.002) with anemia. Candidatus Mycoplasma haematoparvum (30/200, 15%) and Mycoplasma haemocanis (4/200, 2%) in dogs and Candidatus Mycoplasma haemominutum (18/100, 18%) and M. haemofelis (2/100, 2%) in cats were detected. Additionally, 2 dogs were positive for C. burnetii DNA. Coinfections were detected in dogs, with the majority (23/200, 11.5%) including B. henselae and C.M. haematoparvum, followed by Mycoplasma haemocanis and C.M. haematoparvum (2/200, 1%) and B. henselae, CMhp and C. burnetii (2/200, 1%). Haemoplasma infection was high in Egyptian dogs and cats with a high prevalence for zoonotic Bartonella spp. in dogs with anemia, highlighting the need to investigate these agents in the diagnostic algorithm of anemia and to adopt preventive measures to protect both animal and human health.

Co-infection patterns of vector-borne zoonotic pathogens in owned free-ranging dogs in central Chile

We investigated the co-occurrence of the nine of the most relevant canine vector-borne pathogens (CVBP) using conventional and real-time PCR and evaluated risk factors and potential non-apparent haematological alterations associated with co-infection in 111 rural, owned, free-ranging dogs in the Metropolitan Region of Chile.At least one pathogen was detected in 75% of the dogs. DNA of Anaplasma platys (Ap; 36%), Candidatus Mycoplasma haematoparvum (CMhp; 31%), Mycoplasma haemocanis (Mhc; 28%), Trypanosoma cruzi (17%), Leishmania spp. (4.5%), and Acanthocheilonema reconditum (1%) was detected. All dogs were negative for Ehrlichia spp., Rickettsia spp., Bartonella spp., Piroplasmida, and Hepatozoon spp. Thirty-eight dogs (34%) were coinfected. CMhp was involved in 71%, Mhc in 58%, and Ap in 50% of the co-infections. The most common co-infection pattern was CMhp–Mhc (37% of the cases). The prevalence of Ap was higher in juvenile than in adult dogs, whereas the opposite was found for CMhp and Mhc. Adult dogs were four times more likely of being co-infected than juveniles. Co-infected animals showed higher white blood cell count, segmented neutrophil count, and GGT levels than non-co-infected dogs. Clinically healthy but infected dogs may act as reservoirs of CVBP, and their free-ranging behavior would facilitate the spread of these pathogens to other dogs as well as human beings or wild carnivores.

Nanopore Sequencing Using the Full-Length 16S rRNA Gene for Detection of Blood-Borne Bacteria in Dogs Reveals a Novel Species of Hemotropic Mycoplasma.

Dogs across the globe are afflicted by diverse blood- and vector-borne bacteria (VBB), many of which cause severe disease and can be fatal. Diagnosis of VBB infections can be challenging due to the low concentration of bacteria in the blood, the frequent occurrence of coinfections, and the wide range of known, emerging, and potentially novel VBB species encounterable. Therefore, there is a need for diagnostics that address these challenges by being both sensitive and capable of detecting all VBB simultaneously. We detail the first employment of a nanopore-based sequencing methodology conducted on the Oxford Nanopore Technologies (ONT) MinION device to accurately elucidate the "hemobacteriome" from canine blood through sequencing of the full-length 16S rRNA gene. We detected a diverse range of important canine VBB, including Ehrlichia canis, Anaplasma platys, Mycoplasma haemocanis, Bartonella clarridgeiae, "Candidatus Mycoplasma haematoparvum", a novel species of hemotropic mycoplasma, and Wolbachia endosymbionts of filarial worms, indicative of filariasis. Our nanopore-based protocol was equivalent in sensitivity to both quantitative PCR (qPCR) and Illumina sequencing when benchmarked against these methods, achieving high agreement as defined by the kappa statistics (k > 0.81) for three key VBB. Utilizing the ability of the ONT' MinION device to sequence long read lengths provides an excellent alternative diagnostic method by which the hemobacteriome can be accurately characterized to the species level in a way previously unachievable using short reads. We envision our method to be translatable to multiple contexts, such as the detection of VBB in other vertebrate hosts, including humans, while the small size of the MinION device is highly amenable to field use. IMPORTANCE Blood- and vector-borne bacteria (VBB) can cause severe pathology and even be lethal for dogs in many regions across the globe. Accurate characterization of all the bacterial pathogens infecting a canine host is critical, as coinfections are common and emerging and novel pathogens that may go undetected by traditional diagnostics frequently arise. Deep sequencing using devices from Oxford Nanopore Technologies (ONT) provides a solution, as the long read lengths achievable provide species-level taxonomic identification of pathogens that previous short-read technologies could not accomplish. We developed a protocol using ONT' MinION sequencer to accurately detect and classify a wide spectrum of VBB from canine blood at a sensitivity comparable to that of regularly used diagnostics, such as qPCR. This protocol demonstrates great potential for use in biosurveillance and biosecurity operations for the detection of VBB in a range of vertebrate hosts, while the MinION sequencer's portability allows this method to be used easily in the field.

Molecular detection of haemophilic pathogens reveals evidence of Candidatus Mycoplasma haemobos in dogs and parasitic ticks in central China

BackgroundIn addition to Mycoplasma haemocanis and Candidatus Mycoplasma haematoparvum, a few hemoplasma species that mainly infect other livestock have been detected in dogs. ‘Candidatus Mycoplasma haemobos’ (Ca. M. haemobos) has been found in a variety of animals in China. The present study was aimed to investigate the occurrence of ‘Ca. M. haemobos’ infections in dogs and ticks collected from the Henan province, China.ResultsOverall, 55 dog blood samples and 378 ticks on skins were collected from anemic and healthy dogs, and these samples were subjected to PCR, sequence analysis, and identification. The results showed that Haemaphysalis longicornis (266) and Rhipicephalus (Boophilus) microplus (112) were the only two parasitic ticks on dogs. Molecular detection revealed that 163 M. haemocanis, 88 ‘Ca. M. haemobos’ and 32 Anaplasma platys positive amplicons could be amplified from dogs, H. longicornis and R. (B.) microplus. In addition, co-infections (M. haemocanis + A. platys and ‘Ca. M. haemobos’+ A. platys) could be also detected.ConclusionsTo the best of our knowledge, this is the first molecular evidence of ‘Ca. M. haemobos’ natural infection in dogs and tick species identified as H. longicornis and R. (B.) microplus from China.

Expanding the Universe of Hemoplasmas: Multi-Locus Sequencing Reveals Putative Novel Hemoplasmas in Lowland Tapirs (Tapirus terrestris), the Largest Land Mammals in Brazil.

The lowland tapir (Tapirus terrestris) is the largest land mammal in Brazil and classified as a vulnerable species, according to the assessment of the risk of extinction. The present study aimed at investigating the occurrence and genetic diversity of hemoplasmas in free-ranging T. terrestris from the Brazilian Pantanal and Cerrado biomes. Blood samples were collected from 94 living and eight road-killed tapirs, totalizing 125 samples Conventional PCR targeting four different genes (16S rRNA, 23S rRNA, RNAse P, and dnaK) were performed, and the obtained sequences were submitted for phylogenetic, genotype diversity, and distance analyses. The association between hemoplasma positivity and possible risk variables (age, gender, and origin) was assessed. Out of 122 analyzed samples, 41 (41/122; 33.61% CI: 25.84–42.38%) were positive in the 16S rRNA-based PCR assay for hemoplasmas. Positivity for hemoplasmas did not differ between tapirs’ gender and age. Tapirs from Pantanal were 5.64 times more likely to present positive results for hemoplasmas when compared to tapirs sampled in Cerrado. BLASTn, phylogenetic, genotype diversity, and distance analyses performed herein showed that the sampled lowland tapirs might be infected by two genetically distinct hemoplasmas, namely ‘Candidatus Mycoplasma haematoterrestris’ and ‘Candidatus Mycoplasma haematotapirus’. While the former was positioned into “Mycoplasma haemofelis group” and closely related to ‘Candidatus Mycoplasma haematoparvum, the latter was positioned into “Mycoplasma suis group” and closely related to ‘Candidatus Mycoplasma haematobos’. The impact of both putative novel species on tapir health status should be investigated.

First molecular evidence of Mycoplasma haemocanis and ‘Candidatus Mycoplasma haematoparvum’ infections and its association with epidemiological factors in dogs from Cuba

Haemotrophic mycoplasmas (haemoplasmas) are unculturable, epicellular, cell wall-less gram-negative bacteria distributed worldwide, which infect several mammalian species. In dogs, Mycoplasma haemocanis and ‘Candidatus Mycoplasma haematoparvum’ have been reported as causative agents of infectious anaemia, especially in splenectomised or immunocompromised animals. The present cross-sectional study aims to assess the prevalence, risk factors, and molecular characterization of canine haemoplasmas in Cuba. A total of 391 dog blood samples and 247 tick samples were tested for the presence of canine haemoplasmas by species-specific quantitative TaqMan® real-time PCR assays. Overall, 17.9% (70/391; 95% CI: 14.1–21.7) blood samples were PCR-positive for at least one canine haemoplasmas species, where 15.1% (59/391; 95% CI: 11.5–18.7) for Mycoplasma haemocanis, 4.4% (17/391; 95% CI: 2.3–6.4) for ‘Candidatus Mycoplasma haematoparvum’, and 1.5% (6/391; 95% CI: 0.3–2.8) were co-infected. All collected ticks were identified morphologically as Rhipicephalus sanguineus sensu lato, and none of the tested tick samples was found PCR-positive for the presence of Mycoplasma haemocanis and ‘Candidatus Mycoplasma haematoparvum’. Risk factors for canine haemoplasmas species infection included the presence of tick infestation, crossbreeding and living in kennels, while no association was found with the occurrence of anaemia. Phylogenetic analyses based on the 16S rRNA gene sequences of Mycoplasma haemocanis and ‘Candidatus Mycoplasma haematoparvum’ revealed >99% identity to other isolates distributed worldwide, indicating low genetic variability amongst these canine haemoplasmas species. To the best of the authors´ knowledge, this is the first molecular evidence of Mycoplasma haemocanis and ‘Candidatus Mycoplasma haematoparvum’ infections in dogs from Cuba.

A multipronged next-generation sequencing metabarcoding approach unearths hyperdiverse and abundant dog pathogen communities in Cambodia.

Recent surveys in Southeast Asia, including Cambodia, have identified canine vector-borne pathogens (VBPs), including those with zoonotic potential, as highly prevalent. The lack of veterinary care alongside the close association semidomesticated dogs have with humans in the region exacerbates these zoonotic risks. Nonetheless, the number of studies investigating such pathogens and the threats they pose to dog and human health is limited. Here, we utilize a next-generation sequencing (NGS)-based metabarcoding protocol to conduct an assumption-free characterization of the bacterial, apicomplexan, and kinetoplastid blood-borne pathogens of free-roaming dogs from across Cambodia. From 467 dogs at five field sites, 62% were infected with one of eight confirmed pathogens, comprising Anaplasma platys (32%), Ehrlichia canis (20%), Hepatozoon canis (18%), Babesia vogeli (14%), Mycoplasma haemocanis (13%), the zoonotic pathogen Bartonella clarridgeiae (3%), Candidatus Mycoplasma haematoparvum (0.2%), and Trypanosoma evansi (0.2%). Coinfections of between two and four VBPs were common with 28% of dogs found to have a mixed infection. Moreover, DNA from putatively infectious agents belonging to the bacterial family and genera Coxiella, Mycobacterium, Neisseria, Rickettsiaceae, Treponema, and two uncharacterized Mycoplasma species were identified, in addition to protozoan genera Colpodella, Parabodo, and Bodo. Using a multiple logistic regression model, the presence of ectoparasites, abnormal mucous membranes, anemia, and total protein were found as predictors of canine VBP exposure. This study represents the first time an NGS metabarcoding technique has been used to holistically detect the bacterial and protozoan hemoparasites communities of dogs through an in-depth survey, highlighting the power of such methods to unearth a wide spectrum of pathogenic organisms in an unbiased manner.

First molecular detection of Canine Hemoplasmas in Sivas province in central part of Turkey

Canine hemoplasmas are tick-borne bacterial pathogens having worldwide distribution. There are two hemoplasmas species that cause disease in dogs. These are Mycoplasma haemocanis and Candidatus Mycoplasma haematoparvum. The aim of this study was to evaluate the prevalence of canine hemoplasmas among apparently healthy 194 owned-dogs in Sivas Province using species-specific polymerase chain reaction (PCR). According to our results, the overall prevalence of canine hemoplasmas was 14.94% (29/194). The molecular prevalence of M. haemocanis and Ca. M. haematoparvum was detected as 8.24% (16/194) and 10.50% (21/194) respectively while the prevalence of co-infections was 4.12% (8/194). In this study, Ca. M. haematoparvum which can infect humans was found more prevalent than M. haemocanis. To the best of our knowledge, it is the first molecular study on the determination of canine hemoplasmas in Sivas province in Turkey.

Risk of transmittable blood-borne pathogens in blood units from blood donor dogs in Canada.

BackgroundCanine blood donors can be infected by various vector‐borne or other pathogens that could be an important cause of morbidity and death in transfusion recipients.Hypothesis/ObjectivesTo estimate and predict positivity to transmittable blood‐borne pathogens in blood units collected from blood donor dogs in Canada.AnimalsSix thousand one hundred and fifty blood units from 1914 active blood donors registered to the Canadian Animal Blood Bank (CABB) between March 2010 and December 2016.MethodsA registry‐based retrospective study. Blood units were screened by SNAP 4Dx/4Dx Plus and PCR panel tests. Information on blood donors and test results were extracted from multiple databases and collated. Logistic regressions were used to predict blood unit positivity.ResultsOf 1779 blood units, 0.56% were antibody‐positive for Anaplasma phagocytophilum/platys and 0% for Ehrlichia canis/ewingii. After exclusion of antibody‐positive units to Anaplasma spp., 1.1% of 6140 blood units were PCR‐positive to Anaplasma phagocytophilum, Bartonella spp., Brucella canis, “Candidatus Mycoplasma haematoparvum,” Mycoplasma haemocanis, or a combination of these pathogens. Babesia spp., Ehrlichia spp., and Leishmania spp. were not detected. Units from the first blood collection from a dog had higher odds of testing PCR‐positive (P < .001) for at least 1 pathogen than units from subsequent collections.Conclusions and Clinical ImportanceAlthough our study indicates a low probability of detecting blood‐borne pathogen in blood units collected by this Canadian blood bank, the presence of positive units highlights the importance of the preemptive identification and screening of blood units from healthy blood donors for safe blood banking, especially in first‐time donors.

Widespread Infection with Hemotropic Mycoplasmas in Free-Ranging Dogs and Wild Foxes Across Six Bioclimatic Regions of Chile.

Blood samples of 626 rural dogs, 140 Andean foxes (Lycalopex culpaeus), and 83 South American grey foxes (L. griseus) from six bioregions of Chile spanning 3000 km were screened for Mycoplasma DNA by conventional PCR and sequencing. Risk factors of infection were inferred using Generalized Linear Mixed Models and genetic structure by network analyses. Overall, Mycoplasma haemocanis/Mycoplasma haemofelis (Mhc/Mhf) and Candidatus Mycoplasma haematoparvum (CMhp) observed prevalence was 23.8% and 12.8% in dogs, 20.1% and 7.2% in Andean foxes, and 26.5% and 8.4% in grey foxes, respectively. Both hemoplasmas were confirmed in all the bioregions, with higher prevalence in those where ticks from the Rhipicephalus sanguineus species group were absent. Candidatus M. haematominutum and a Mycoplasma sp. previously found in South American carnivores were detected in one fox each. Although the most prevalent Mhc/Mhf and CMhp sequence types were shared between dogs and foxes, network analysis revealed genetic structure of Mhc/Mhf between hosts in some regions. Male sex was associated with a higher risk of Mhc/Mhf and CMhp infection in dogs, and adult age with CMhp infection, suggesting that direct transmission is relevant. No risk factor was identified in foxes. Our study provides novel information about canine hemoplasmas with relevance in distribution, transmission routes, and cross-species transmission.

Illuminating the bacterial microbiome of Australian ticks with 16S and Rickettsia-specific next-generation sequencing

Next-generation sequencing (NGS) studies show that mosquito and tick microbiomes influence the transmission of pathogens, opening new avenues for vector-borne pathogen control. Recent microbiological studies of Australian ticks highlight fundamental knowledge gaps of tick-borne agents. This investigation explored the composition, diversity and prevalence of bacteria in Australian ticks (n = 655) from companion animals (dogs, cats and horses). Bacterial 16S NGS was used to identify most bacterial taxa and a Rickettsia-specific NGS assay was developed to identify Rickettsia species that were indistinguishable at the V1-2 regions of 16S. Sanger sequencing of near full-length 16S was used to confirm whether species detected by 16S NGS were novel. The haemotropic bacterial pathogens Anaplasma platys, Bartonella clarridgeiae, “Candidatus Mycoplasma haematoparvum” and Coxiella burnetii were identified in Rhipicephalus sanguineus (s.l.) from Queensland (QLD), Western Australia, the Northern Territory (NT), and South Australia, Ixodes holocyclus from QLD, Rh. sanguineus (s.l.) from the NT, and I. holocyclus from QLD, respectively. Analysis of the control data showed that cross-talk compromises the detection of rare species as filtering thresholds for less abundant sequences had to be applied to mitigate false positives. A comparison of the taxonomic assignments made with 16S sequence databases revealed inconsistencies. The Rickettsia-specific citrate synthase gene NGS assay enabled the identification of Rickettsia co-infections with potentially novel species and genotypes most similar (97.9–99.1%) to Rickettsia raoultii and Rickettsia gravesii. “Candidatus Rickettsia jingxinensis” was identified for the first time in Australia. Phylogenetic analysis of near full-length 16S sequences confirmed a novel Coxiellaceae genus and species, two novel Francisella species, and two novel Francisella genotypes. Cross-talk raises concerns for the MiSeq platform as a diagnostic tool for clinical samples. This study provides recommendations for adjustments to Illuminaʼs 16S metagenomic sequencing protocol that help track and reduce cross-talk from cross-contamination during library preparation. The inconsistencies in taxonomic assignment emphasise the need for curated and quality-checked sequence databases.

Babesia gibsoni emerging with high prevalence and co-infections in “fighting dogs” in Hungary

Babesia gibsoni is considered as an emerging protozoan parasite of dogs in North America and Europe. However, no data have been published on its prevalence, molecular-phylogenetic characteristics and associated co-infections in dogs used for illegal fighting (i.e. predisposed to acquiring this piroplasm via biting) in Europe. In this study, blood samples from 79 American Staffordshire Terrier dogs, confiscated for illegal dog fights, were molecularly analyzed for tick-borne pathogens. Babesiagibsoni was detected in 32 dogs, i.e. with a prevalence of 40.5%. In addition, Babesia vulpes was found in 8 samples (prevalence of 10.1%), for the first time in dogs in Hungary. Canine hemoplasmas were also identified in 49 samples (62%): only Mycoplasma haemocanis in 32 (40.5%) dogs, only “Candidatus Mycoplasma haematoparvum” in 9 (11.4%) dogs, and both hemoplasmas in 8 (10.1%) dogs. Thus, hemoplasma infections also showed a particularly high prevalence in this dog population. Based on a partial fragment of the 18S rRNA gene, B. gibsoni from Hungary exhibited complete sequence identity with conspecific strains reported from Europe and Asia. The cytochrome c oxidase subunit 1 (cox1) gene sequence of this isolate showed the closest identity with B. gibsoni reported from Japan but had a nonsynonymous mutation (M33I). Furthermore, the 11 B. gibsoni-positive samples analyzed for sequence variants of the cytochrome b (cytb) gene showed the presence of a common mutation (P310S). Most importantly, B. gibsoni had two further nonsynonymous mutations, M121I and F258L, in a dog with severe and relapsing anemia following atovaquone treatment. Phylogenetically, both cytb sequence variants clustered together, with a clear geographical pattern showing the closest relationship of both haplotypes identified in Hungary with those from China and Japan. To the best of our knowledge, this is the first cox1 and cytb characterization of B. gibsoni in Europe, as well as the first report on the emergence of this piroplasm and hemoplasmas with high prevalence among “fighting dogs” north of the Mediterranean Basin.

Distribution and risk factors of canine haemotropic mycoplasmas in hunting dogs from southern Italy

Mycoplasma haemocanis (Mhc) and “Candidatus Mycoplasma haematoparvum” (CMhp) are the main haemoplasma species known to infect dogs. The aim of this study was to determine the prevalence of haemoplasma species infections in hunting dogs from southern Italy and assess related risk factors. 1,433 hunting dogs living in Campania region were tested by qPCR assay. The prevalence was 19.9 %; 13.1 % for Mhc and 11.4 % for CMhp; 4.6 % showed a coinfection with both haemoplasma species. Statistical analysis revealed living in Salerno province (Mhc: OR 3.72; CMhp: OR 2.74), hound (Mhc: OR 5.26; CMhp: OR 8.46) and mixed breed (Mhc: OR 3.38; CMhp: OR 2.80), rural environment (Mhc: OR 12.58; CMhp: OR 10.38), wild mammal hunting (Mhc: OR 8.73; CMhp: OR 8.32), cohabitation with other animals (Mhc: OR 2.82; CMhp: OR 2.78) and large pack size (Mhc: OR 2.96; CMhp: OR 1.61) as risk factors for haemoplasmas. Male gender (OR 1.44) and tick infestation history (OR 1.40) represented risk factors only for Mhc, while adult age (2−7 years - OR 2.01; > 7 years - OR 1.84) and large body size (OR 1.48) were associated only to CMhp. Mhc infection was significantly associated to Babesia vogeli (p < 0.05) and Hepatozoon canis (p < 0.001), while CMhp with H. canis (p < 0.001). This study adds information on haemoplasma species distribution in hunting dogs in southern Italy. Outdoor lifestyle and contact with wild fauna, through greater exposure to tick infestation, or possibly wounds acquired during hunting or fighting, could be factors contributing to haemoplasma infections.

Molecular survey of Bartonella spp. and haemoplasmas in American minks (Neovison vison).

The aim of this study was to perform a molecular survey and characterize Bartonella spp. and haemotropic Mycoplasma (haemoplasmas) in invasive American minks (Neovison vison) from Southern Chile. Additionally, we addressed risk factors for positivity in both groups of agents. Blood and/or tissue samples from 246 minks were analysed by qPCR targeting the nuoG gene for Bartonella spp. and conventional (c)PCR for 16S rRNA for haemotropic Mycoplasma spp. nuoG qPCR-positive Bartonella spp. samples were submitted to cPCR assays (ITS, ribC, gltA, rpoB, pap-31 and ftsZ genes) to perform phylogenetic inferences. Haemotropic Mycoplasma spp. 16S-positive samples were further amplified by cPCR targeting RNaseP gene (160-210bp) and by two overlapping 16S rRNA cPCR assays to amplify a larger portion of the gene (1,200bp) for phylogenetics. Bartonella DNA was detected in 8.9% of minks (22/246). Out of 22 nuoG qPCR-positive samples, one and two showed positive results in cPCR assays based on ITS and ribC, respectively. Consistent sequencing results were obtained for only one ITS sample (464bp sequence), which shared 99.6% identity with B.clarridgeiae. Two per cent of minks (5/246) were positive for 16S rRNA haemotropic Mycoplasma-cPCR assay. Two concatenated sequences of 16S rRNA (1,176 and 1,230bp) were obtained: one sample shared 97.87% identity with haemotropic Mycoplasma sp. from a wild rodent, and the other 96.49% identity with 'Candidatus Mycoplasma haematoparvum' from a dog. All BLAST results were supported by phylogenetic analysis. One haemoplasma RNase P sequence shared 94.86% identity with Mycoplasma haemofelis from a cat. No risk factors for PCR positivity were identified. In a nutshell, Bartonella clarridgeiae and a potentially novel haemoplasma closely related to haemoplasmas previously reported in rodents, dogs, domestic and wild cats were described for the first time in American minks.

Immune-Mediated Hemolytic Anemia Associated with Candidatus Mycoplasma Haematoparvum in a Splenectomized Dog in Italy

Abstract This report describes a case of canine hemotropic mycoplasmasosis by Candidatus Mycoplasma haematoparvum in a dog. A five-year-old splenectomized dog was referred to the Veterinary Teaching Hospital of the University of Sassari with clinical symptoms and laboratory findings compatible with immune-mediated hemolytic anemia. Epicellular bacteria were detected in the erythrocytes by microscopic examination of blood smears. PCR and sequencing were positive for Candidatus Mycoplasma haematoparvum. Treatment with doxycycline, prednisolone and blood transfusion was administered. Several studies have described the molecular prevalence of M. hemocanis and Candidatus M. haematoparvum, however there are few clinical reports, especially those describing Candidatus M. haematoparvum infection in dogs, for which only two cases have been reported. To the best of our knowledge this is the first case report of a symptomatic infection caused by Candidatus Mycoplasma haematoparvum in Italy. Hemoplasmosis should be considered as a potential cause of hemolytic anemia in dogs. Following treatment with doxycycline and prednisolone, the clinical signs improved without resolution of infection. This condition was the same at the three-year follow-up.

Hemoplasma infection in wild captive carnivores

ABSTRACT: Hemoplasmas are bacteria able to adhere themselves loosely to the plasma membrane of erythrocytes and may parasitize several species of mammals. There are three known species of hemoplasmas that parasitize domestic and wild cats: Mycoplasma haemofelis, ‘Candidatus Mycoplasma haemominutum’ and ‘Candidatus Mycoplasma turicensis’. Dogs are infected by at least two species of hemoplasmas: ‘Candidatus Mycoplasma haematoparvum’ and Mycoplasma haemocanis. The hemoplasmoses are very important in veterinary clinics, either because of its worldwide distribution and severity of clinical signs, depending on parasite species and host immune competence, or due to its zoonotic potential and capability of infecting endangered species. This study set out to investigate which hemoplasmas species parasitize different captive wild carnivores in order to clarify the epidemiology of hemoplasmoses in wild animals. Furthermore, the research intended to characterize the hematological changes caused by different species of hemotropic mycoplasmas infection in order to establish their clinical importance to wild species and the capacity of these species to become a reservoir of studied agents. Samples of 33 wild felids and 18 wild canids were investigated using polymerase chain reaction (PCR) to detect hemoplasmas DNA and it was observed that the occurrence of infection in these species is 45.5% and 83.3%, respectively. Factors such as age, gender or anaemia are not more frequent in animals positive for the infection. Therefore, it is concluded that infection caused by hemoplasmas in wild carnivores has high prevalence, and either agent pathogenicity is low, or chronic stage is more frequent, resulting in a low rate of diagnosis.

Report of Candidatus Mycoplasma haematoparvum and Mycoplasma haemocanis canine infections in Massambaba restinga, Brazil

Tick-borne diseases are frequent in the southeastern section of Brazil. The most prevalent canine parasites diagnosed are Ehrlichia canis, Babesia gibsoni, Babesia canis, and Anaplasma platys, although Mycoplasma haemocanis and Candidatus Mycoplasma haematoparvum have also been detected in other regions of the country. Two clinically healthy dogs from a suburban area of the state of Rio de Janeiro had a history of being heavily infested with ticks and were examined at IDEXX Reference Laboratories, California for a tick panel check. One dog harbored DNA of Candidatus Mycoplasma haematoparvum and the other, the DNA of Mycoplasma haemocanis. These results reinforce the need for permanent monitoring for tick infestations and tick-borne parasites in southeastern Brazil, particularly considering the likely continuation of global climate changes that will contribute to the spread and increase of infections across the country.

Tick-borne infections of dogs in Serbia: A review of research

Increased prevalence of tick-borne disease in humans and animals is an important public health issue. Dogs can be clinically ill, reservoirs of tick-borne pathogens, or sentinel populations for research. Certainly, small-scale research has been conducted in dogs and this review will focus on the recent studies of emergent and re-emergent tick-borne pathogens in dogs in Serbia. Babesiosis stands out as the most common and important tick-borne disease in dogs. In addition to this pathogen, the presence of Hepatozoon canis and ?Candidatus Mycoplasma haematoparvum? in the blood of dogs was proved using molecular methods. Seroreactivity in dogs has been proven for Borrelia burgdorferi sensu lato (s.l.) complex, Ehrlichia canis, Ehrlichia ewingii, Anaplasma phagocytophilum, Anaplasma platys, Rickettsia conorii, and tick-borne encephalitis virus (TBEV). The presence of B. burgdorferi s.l. complex, A. phagocytophilum, Rickettsia spp. from the group of spotted fevers, and TBEV in ticks indicates the geographical distribution and potential for infecting humans, dogs, and other animals in Serbia. Further systematic, comprehensive, well designed, bacteriological, virological, parasitological, epidemiological, clinical (infectological), and acarological research is needed in Serbia.