Candidatus Mycoplasma Haematoparvum Research Articles

Detection of haemotropic Mycoplasma (Haemobartonella) using multiplex PCR and its relationship with epidemiological factors in dogs

The aim of the present study was to conduct multiplex PCR for detection of two haemotropic Mycoplasma species in dogs from central Iran and study their relationship with epidemiological factors. In order to achieve this, blood samples from 100 Esfahan dogs were analysed by PCR to evaluate haemoplasma infection status. Thirteen were infected with an organism closely related to Mycoplasma haemocanis, and ten were infected with Candidatus Mycoplasma haematoparvum, whilst three dogs were contaminated with both organisms. The results showed a significant difference between young (1–5 years old) and ectoparasite-infected dogs for the presence of haemotropic Mycoplasma. Our survey indicates that multiplex PCR assay is able to successfully detect haemotropic Mycoplasma in dogs.

Candidatus Mycoplasma haematoparvum and Mycoplasma haemocanis infections in dogs from the United States

Mycoplasma haemocanis (Mhc) and Candidatus Mycoplasma haematoparvum (CMhp) have been described in dogs. Historically, microscopic visualization of hemotropic Mycoplasma spp. has occurred most often in immunocompromised or splenectomized dogs. The aim of this study was to determine the Mhc and CMhp prevalences among dogs from the United States. Novel 16S rRNA and RNAseP gene PCR assays were used to amplify hemotropic Mycoplasma species DNA for GenBank sequence alignment. Among the study population, hemoplasma prevalence was 1.3% (7 out of 506), with Mhc and CMhp prevalences of 0.6% and 0.8%, respectively. Two of six CMhp-infected dogs were co-infected with a Bartonella sp., and a third dog was seroreactive to Bartonella henselae antigens. The prevalence of Mhc and CMhp in this study was low; potential blood donors should be screened; and dogs and people can be co-infected with hemoplasma and Bartonella spp.

Prevalence of canine haemotropic mycoplasma infections in Sydney, Australia

Haemotropic mycoplasmas (haemoplasmas) are epierythrocytic, wall-less bacteria that cannot be cultivated in vitro (Messick 2003). Two species have been identified in dogs: Mycoplasma haemocanis (Mhc) and ‘ Candidatus Mycoplasma haematoparvum’...

Canine vector‐borne disease pathogens in dogs from south‐east Queensland and north‐east Northern Territory

To determine the prevalence of canine vector-borne diseases (CVBD: Babesia spp., Anaplasma spp., Ehrlichia spp., haemotropic mycoplasmas and Hepatozoon) in Australian dogs; namely, dogs from pounds in south-east Queensland and an indigenous Aboriginal community in the north-east of the Northern Territory. Blood samples were collected from 100 pound dogs and 130 Aboriginal community dogs and screened for the CVBD pathogens using polymerase chain reaction (PCR). All positive PCR products were sequenced for species confirmation. In total, 3 pound dogs and 64 Aboriginal community dogs were infected with at least one CVBD pathogen. Overall, B. vogeli was detected in 13 dogs, A. platys in 49, M. haemocanis in 23, Candidatus Mycoplasma haematoparvum in 3 and C. M. haemobos in 1 dog. Co-infections were detected in 22 Aboriginal community dogs. This study found B. vogeli, A. platys and haemotropic mycoplasma infections to be common in dogs in subtropical and tropical areas of Australia. This study also reports for the first time the prevalence and genetic characterisation of haemotropic mycoplasmas in dogs in Australia.

Haemoparasites of free-roaming dogs associated with several remote Aboriginal communities in Australia

BackgroundTick-borne haemoparasites Babesia vogeli and Anaplasma platys are common among the free-roaming canine populations associated with Aboriginal communities in Australia, whilst the prevalence of haemoplasmas, which are also suspected to be tick-borne, remained unexplored. The aim of this study was to determine the prevalence of haemoplasma infection in these populations, and to identify any correlation with other haemoparasites. Blood was collected from 39 dogs associated with four Aboriginal communities and screened for infection using PCR and serology. DNA was purified and PCR analyses for piroplasms, Anaplasmataceae family bacteria and haemoplasmas performed. Serum was analysed using a commercial haemoparasite ELISA. Prevalence of infection was compared between communities.ResultsSeventeen dogs (44%) were infected (PCR positive) with Mycoplasma haemocanis, eight (21%) with ‘Candidatus Mycoplasma haematoparvum’, 20 (51%) with A. platys, and 17 (44%) with B. vogeli. Two dogs were infected with a novel haemoplasma as determined by DNA amplification and sequencing. Two dogs (5%) were serologically positive for Dirofilaria immitis antigens, one (3%) was positive for Ehrlichia canis antibodies and nine (24nbsp;%) were positive for A. platys antibodies. Co-infections were frequent. Haemoplasma prevalence was highest (73%, 16/22) in Central Australia and lowest (22%, 2/9) in Western Australia (p = 0.017). In contrast, B. vogeli prevalence was low in Central Australia (18%, 4/22) but higher (78%, 7/9) in Western Australia (p = 0.003).ConclusionsThis is the first time haemoplasma infections, including a novel species, have been molecularly documented in Australian dogs. The wide regional variation in prevalence of some of the haemoparasite infections detected in this study warrants further investigation.

Real‐time quantitative polymerase chain reaction detection of haemoplasmas in healthy and unhealthy dogs from Central Macedonia, Greece

The aims of this study were to determine the prevalence of canine haemoplasmas, Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum" infection in Central Macedonia, Greece, and to evaluate any associations between canine haemoplasma infection and clinical presentation, selected laboratory data or the presence of ticks. Genomic DNA was purified from excess blood (n=151) submitted for haematological examination. Purified DNA was subjected to species-specific quantitative polymerase chain reaction assays duplexed with a canine DNA control quantitative polymerase chain reaction. Clinical records were retrospectively examined and selected clinical parameters were compared to haemoplasma infection status. Nine samples were excluded due to inadequate canine DNA polymerase chain reaction results. Of the remaining 142 samples: eight (5·6%) were positive for M. haemocanis alone, six (4·2%) were positive for "Ca. M. haematoparvum" alone and one (0·7%) was dual positive. No association was found between haemoplasma status and age, sex, breed, health status, presence of anaemia, selected biochemistry parameters, presence of ectoparasites, routine ectoparasiticide treatment or the presence of selected tick-borne diseases.

Hemoplasmas in Wild Canids and Felids in Brazil

Hemotropic mycoplasmas, epicellular erythrocytic bacterial parasites lacking a cell wall, are the causative agents of infectious anemia in numerous mammalian species. The presence of hemotropic mycoplasmas in blood samples of neotropical and exotic wild canids and felids from Brazilian zoos were recorded using molecular techniques. Blood samples were collected from 146 Brazilian wild felids, 19 exotic felids, 3 European wolves (Canis lupus), and from 97 Brazilian wild canids from zoos in the Brazilian states of São Paulo and Mato Grosso and the Federal District. Using conventional polymerase chain reaction (PCR), this work found 22 (13%) wild felids positive to Candidatus Mycoplasma haemominutum [4 jaguars (Panthera onca); 3 pumas (Puma concolor); 10 ocelots (Leopardus pardalis); 2 jaguarondis (Puma yagouaroundi); and 3 little spotted cats (Leopardus tigrinus)]. Only one little spotted cat (Leopardus tigrinus) was positive to Mycoplasma haemofelis, and none was positive to Candidatus Mycoplasma turicensis. Two bush dogs (Speothos venaticus) were positive for a Mycoplasma sp. closely related to Candidatus Mycoplasma haematoparvum, and two European wolves were positive for a Mycoplasma sp. closely related to Candidatus Mycoplasma haemominutum. This is the first study regarding the molecular detection of hemotropic mycoplasmas in wild canids.

Haemoplasma infection is not a common cause of canine immune-mediated haemolytic anaemia in the UK

The aim of this study was to investigate whether the two canine haemoplasma species, Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum," are commonly associated with immune-mediated haemolytic anaemia (IMHA) in UK dogs. Three groups of dogs were recruited to the study: anaemic dogs with primary IMHA (n=37); anaemic dogs not meeting the inclusion criteria for primary IMHA (n=77) and non-anaemic dogs (n=113). DNA was extracted from 100 μl of blood and subjected to real-time quantitative polymerase chain reaction (qPCR) assays for both species of Mycoplasma. Each assay incorporated co-amplification of canine glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous internal control. Canine GAPDH was successfully amplified by qPCR from all 227 canine blood samples but none contained M. haemocanis or "Candidatus M. haematoparvum" DNA. Haemoplasma infection is uncommon in dogs in the UK and no evidence was found that these organisms act as triggers for IMHA.

Haemotrope Mykoplasmen bei Hund und Katze: Übertragung, Diagnose, Prävalenz und Bedeutung in Europa

Haemotropic mycoplasmas (or haemoplasmas) are the causative agents of infectious anaemia in many mammalian species. They were previously known as Haemobartonella and Eperythrozoon species. The development of sensitive, specific PCR assays has expanded our knowledge of these agents and PCR is the method of choice to diagnose and differentiate haemoplasma infections. In felids, Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis' have been described. They vary strongly in their pathogenic potential and co-factors may influence the disease severity. In dogs, Mycoplasma haemocanis and 'Candidatus Mycoplasma haematoparvum' are known; clinical signs are mainly found in immunocompromised dogs. Transmission of haemoplasmas may occur via infected blood (aggressive interaction, transfusion) or blood-sucking arthropods. Infections can be treated with Doxycycline, although it is disputable whether the infection is completely eliminated. Feline haemoplasmas must be expected in cats all over Europe, while canine haemoplasmas are mainly encountered in dogs in Mediterranean countries but should also be considered in Swiss dogs with a travel history.

Prevalence of Hemotropic Mycoplasmas in Healthy and Unhealthy Cats and Dogs in Spain

The aims of the present study were to determine the prevalence of hemoplasmas in cats and dogs from the Barcelona area of Spain with the use of species-specific quantitative polymerase chain reaction (qPCR) assays and to evaluate any associations between hemoplasma infection, clinical presentation, and vector-borne infections. Blood samples from cats (191) and dogs (182) were included and were classified as healthy (149) or unhealthy (224). Ethylenediamine tetra-acetic acid blood samples underwent DNA extraction and qPCR analysis. Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum', and 'Candidatus Mycoplasma turicensis' were detected in cats, whereas Mycoplasma haemocanis and 'Candidatus Mycoplasma haematoparvum' were detected in dogs, with prevalences of 3.7%, 9.9%, 0.5%, 14.3%, and 0.6%, respectively. In cats, no association between hemoplasma infection and health status, age, breed, presence of anemia, Feline leukemia virus status, and other vector-borne infections was found, but outdoor access (P = 0.009), male sex (P = 0.01), and Feline immunodeficiency virus status (P < 0.001) were significantly associated with hemoplasma infection. In dogs, sex, age, health status, presence of anemia, and breed were not significantly associated with hemoplasma infection, but a significant association was found between hemoplasma infection and vector-borne infections (P < 0.001). The present report documents the occurrence of feline 'Candidatus M. turicensis' and canine 'Candidatus M. haematoparvum' infections in Spain.

First report about canine infections with Candidatus Mycoplasma haematoparvum in Serbia

There are two species of haemotropic mycoplasmas that infect dogs which are known so far. These are Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum". In the case of dogs, the clinical picture of these infections manifests itself in immunocompromised dogs, although the organisms are also present in immunocompetent dogs which do not show any signs of infection. The vector for these microorganisms is the tick Rhipicephalus sanguineus. The goal of this research was to confirm the presence of canine infections with haemotropic mycoplasmas in our geographic area, regarding the facts that the vector of canine haemotropic mycoplasmas, the Rhipicephalus sanguineus is widely spread in nature, and that the epidemic indications and the clinical signs of canine infections are also present. By means of light microscopic examination of stained peripheral canine blood smears, scanning electron microscopic examination of canine erythrocytes, as well as PCR and sequence analysis of the genome, the presence of canine infections with "Candidatus Mycoplasma haematoparvum" in our geographic area has been identified for the first time.

Prevalence and geographical distribution of canine hemotropic mycoplasma infections in Mediterranean countries and analysis of risk factors for infection

Two hemoplasma species are known in dogs: Mycoplasma haemocanis (Mhc) and ‘Candidatus Mycoplasma haematoparvum’ (CMhp). Although their transmission routes are poorly understood, Rhipicephalus sanguineus has been suggested as a potential tick vector. The aim of the present study was to assess the prevalence, risk factors, and clinical importance of canine hemoplasmas in countries with a Mediterranean climate where R. sanguineus is highly prevalent using TaqMan real-time PCR, and to molecularly characterize the identified isolates. DNA (canine glyceraldehyde-3-phosphate dehydrogenase) was successfully amplified from all samples collected from 850 dogs in Italy, Spain, and Portugal, and 82 (9.6%) were PCR-positive for canine hemoplasmas (43 Mhc, 34 CMhp and 5 co-infected). The hemoplasma sample prevalence was significantly higher in Portugal (40%) than in Italy (9.5%) and Spain (2.5%). Risk factors for infection included living in kennels, young age, crossbreeding, and mange infection. No association was found with anemia. Phylogenetic analyses of the 16S rRNA and RNase P genes revealed >99% identity to other European isolates. In conclusion, canine hemoplasma infections were readily encountered in Mediterranean countries. The climate and living conditions seemed to influence canine hemoplasma prevalence. The clinical importance of canine hemoplasma infections appeared to be low, but the infection stage of the presented dogs was unknown.

Development and use of real-time PCR to detect and quantify Mycoplasma haemocanis and “Candidatus Mycoplasma haematoparvum” in dogs

Two canine haemoplasma species have been recognised to date; Mycoplasma haemocanis (Mhc), which has been associated with anaemia in splenectomised or immunocompromised dogs, and “Candidatus Mycoplasma haematoparvum” (CMhp), recently described in an anaemic splenectomised dog undergoing chemotherapy. The study aim was to develop quantitative real-time PCR assays (qPCRs) incorporating an endogenous internal control to detect Mhc and CMhp and to apply these assays to DNA samples extracted from canine blood collected in Northern Tanzania (n=100) and from dogs presented to a Trinidadian veterinary hospital (n=185).QPCRs specific for Mhc and CMhp were designed using 16S rRNA gene sequence data, and each was duplexed with an assay specific for canine glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The assays detected ≤10 copies of a sequence-specific haemoplasma plasmid per reaction and neither assay showed cross-reactivity with 106 copies of the sequence-specific plasmid from the non-target canine haemoplasma species.Nineteen of the 100 Tanzanian samples (19%) were positive for Mhc alone and one (1%) was dually infected. One Trinidadian sample was negative for canine GAPDH DNA and was excluded from the study. Of the 184 remaining Trinidadian samples, nine (4.9%) were positive for Mhc alone, five (2.7%) for CMhp alone, and two (1.1%) dually infected.This is the first report of canine haemoplasma qPCR assays that use an internal control to confirm the presence of amplifiable sample DNA, and their application to prevalence studies. Mhc was the most commonly detected canine haemoplasma species.

A review of the occurrence of hemoplasmas (hemotrophic mycoplasmas) in Brazil

Recent studies have been conducted in Brazil using molecular techniques for the detection of hemotrophic mycoplasmas in several mammals. In domestic cats, Mycoplasma haemofelis, "Candidatus M. haemominutum", and "Candidatus M. turicensis" infections have been identified. These species have also been found in free-ranging and captive neotropical felid species. Two canine hemoplasmas, Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum", have been identified in dogs. In commercial swine populations, Mycoplasma suis was found to be highly prevalent, especially in sows. Moreover, novel mycoplasma species have been identified in Brazilian commercial pigs and domestic dogs. A hemoplasma infection in a human patient infected with the human immunodeficiency virus (HIV) was also recently documented. In conclusion, hemoplasma species are common and important infectious agents in Brazil. Further studies should be conducted to better understand their impact on pets, production animals, and wildlife fauna, as well as their role as zoonotic agents, particularly in immunocompromised patients.

The occurrence of the feline “Candidatus Mycoplasma haemominutum” in dog in China confirmed by sequence-based analysis of ribosomal DNA

In the present study, polymerase chain reaction (PCR) assay was used to detect haemoplasmas (haemotropic bacteria) in 40 clinically healthy pet dogs in Foshan city, Guangdong Province, China, and one dog was found positive. Comparison of its 16S ribosomal DNA (rDNA) sequence with relevant sequences showed that the isolated haemoplasma had greater sequence identity to feline species "Candidatus Mycoplasma haemominutum" (99%) than to "Candidatus Mycoplasma haematoparvum" (95%). This result, for the first time, indicates the presence of the feline "Candidatus Mycoplasma haemominutum" in Chinese dogs and it represents the first survey of its kind in China by using PCR assay. The results indicated that dog may represent one of the hosts for the feline "Candidatus Mycoplasma haemominutum".

Real-time PCR-based prevalence study, infection follow-up and molecular characterization of canine hemotropic mycoplasmas

Hemotropic mycoplasmas (hemoplasmas) have been reported in several mammalian species including dogs. Infections may lead to hemolytic anemia, but investigations in the dog had been hampered by the lack of adequate diagnostic methods. Only recently sensitive PCR-based assays were reported for the two canine hemoplasmas, Mycoplasma haemocanis and Candidatus Mycoplasma haematoparvum. By applying these assays, 15.4% of 460 dogs from the south of France tested hemoplasma positive. It was hypothesized that this high prevalence may be associated with the presence of Rhipicephalus sanguineus, a proposed vector for canine hemoplasmas. To address this hypothesis and expand the PCR-based knowledge on canine hemoplasmosis, we investigated dogs in a climatic zone that does not allow for the permanent establishment of R. sanguineus. Blood samples were collected throughout a year from 889 dogs in Switzerland: 1.2% of the dogs tested real-time PCR positive. The infection status was not significantly associated with anemia, age or gender. Phylogenetic analyses of Candidatus M. haematoparvum and M. haemocanis isolates revealed ≥99.8% identity to published sequences. All samples collected from three infected dogs throughout a follow-up period of ≤13 months tested PCR positive. Interestingly, the majority of the infected dogs either had been imported from or had visited regions where R. sanguineus is indigenous. Thus, canine hemoplasma prevalence was found to be low in a country with a climate incompatible with frequent occurrence of R. sanguineus. Nonetheless, veterinarians may expect hemoplasma infections in dogs with a travel history and/or after potential tick vector exposure.

Use of Conventional and Real-Time Polymerase Chain Reaction to Determine the Epidemiology of Hemoplasma Infections in Anemic and Nonanemic Cats

The goals of this study were to develop and apply conventional (c) and real-time TaqMan polymerase chain reaction (PCR) assays for Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma haematoparvum' (Mhp), and 'Candidatus Mycoplasma haemominutum' (Mhm) to blood samples of cats to determine the epidemiology of these infections in cats. Cats are infected with >2 hemoplasma species, and organism load correlates with disease induced by these organisms. Blood samples from 263 anemic and nonanemic cats were used. A retrospective study was conducted. Forty-seven (18%) samples were positive. Three samples (1%) yielded 170 base pair cPCR products, 1 of which was positive for Mhf using real-time PCR. Forty-four samples (17%) yielded 193 base pair cPCR products, 40 of which were positive for Mhm using real-time PCR. Organism loads ranged from 375 X 10(6)/mL to 6.9 x 10(6)/mL of blood. Sequencing of cPCR products from samples testing negative using real-time PCR identified 2 Mhp-like sequences, 1 Mhm-like sequence, and 1 sequence resembling 'Candidatus Mycoplasma turicensis'. Cats infected with Mhm were less likely to be anemic than uninfected cats. Older age, outdoor exposure, feline immunodeficiency virus (FIV) seropositivity, cutaneous squamous cell carcinoma (SCC), and stomatitis were associated with Mhm infection. Cats from the Sacramento Valley were more often infected with Mhm than cats from the San Francisco bay area. Cats may be infected with 4 hemoplasma species. The association between Mhm infection, FIV, and SCC may reflect outdoor roaming status of infected cats. The clustered distribution of infection suggests an arthropod vector in transmission.

犬糸状虫症感染予防に来院した犬のバベシア, ヘモバルトネラおよびエールリッビア感染状況調査

Infections with Babesia gibsoni, haemobartonella and Ehrlichia in 536 dogs was examined using a polymerase chain reaction (PCR) specific for each agent in an animal hospital in Sumoto, an endemic area of canine B. gibsoni infection. Ten of 536 dogs (1.9%) tested positive for B. gibsoni, another 16 (3.0%) tested positive for haemobartonella, and none tested positive for Ehrlichia. No mixed infection of B. gibsoni and haemobartonella was detected. Of 16 dogs testing positive for haemobartonella, five were found to be infected with Mycoplasma haemocanis and the other 11 were infected with Candidatus Mycoplasma haematoparvum.

‘Candidatus Mycoplasma haematoparvum’, a novel small haemotropic mycoplasma from a dog

A novel small haemoplasma was detected following cytological examination of blood smears from a splenectomized dog with haemic neoplasia. The 16S rRNA and rnpB genes of the organism were partially sequenced and a phylogenetic tree constructed. The organism was most closely related to the small feline haemoplasma, 'Candidatus Mycoplasma haemominutum' (94 % 16S rRNA gene nucleotide sequence identity; 75 % rnpB) and was only distantly related to Mycoplasma haemocanis (78 % 16S rRNA gene nucleotide sequence identity; 65 % rnpB). As this organism has not been cultured in vitro, the candidate species name 'Candidatus Mycoplasma haematoparvum' is proposed.