Prevalence Of Haemoplasmas Research Articles

A case of mistaken identity: a systematic review, meta-analysis, and reinvestigation of hemotropic Mycoplasma spp. infection in Ctenocephalides felis (cat flea)

Background Feline-associated hemotropic Mycoplasma (hemoplasmas) are believed to be transmitted by two primary mechanisms: (1) direct transmission via fighting and (2) vector-borne transmission by the cat flea (Ctenocephalides felis). While the efficiency of transmission by C. felis appears low, most manuscripts focus on the prevalence of hemoplasmas in wild-caught fleas and report either a very low (< 3%) or a high (> 26%) prevalence. Therefore, we aimed to assess the influence of sample processing and PCR methods on C. felis hemoplasma infection prevalence. MethodsA systemic review of PubMed articles identified 13 manuscripts (1,531 fleas/flea pools) that met the inclusion criteria (performed PCR for >1 hemoplasma on C. felis collected from cats). Risk of bias was assessed utilizing the ROBINS-E tool. Meta-analysis performed in R of these manuscripts found that not washing samples and a common set of 16S rRNA primers first published in Jensen et al. 2001 were associated with increased hemoplasma prevalence. To evaluate the influence of washing on newly collected fleas, we assessed the hemoplasma status of 20 pools of 5 C. felis each, half of which were washed and half not washed.ResultsFlea washing did not influence the detection of hemoplasma but instead amplified Spiroplasma. To assess non-specific amplification with the Jensen et al. 2001 primers, 67 C. felis samples (34% previously reported hemoplasma infected) were subject to PCR and sequencing. By this method, hemoplasma was detected in only 3% of samples. In the remaining “hemoplasma infected” fleas, PCR amplified Spiroplasma or other bacteria. ConclusionsTherefore, we concluded that hemoplasma infection in C. felis is rare, and future flea prevalence studies should sequence all positive amplicons to validate PCR specificity. Further investigation of alternative methods of feline-associated hemoplasma transmission and the ability of C. felis to maintain hemoplasma infection is necessary.Graphical

Preliminary report of Mycoplasma Wenoynii and Candidatus Mycoplasma haemobos infection in Korean native cattle

BackgroundHemotropic mycoplasmas or hemoplasmas are bacteria that attach to the erythrocyte surface and cause bovine hemoplasmosis. Two species, Mycoplasma wenyonii and Candidatus Mycoplasma haemobos, have been identified and shown to be distributed worldwide. However, there is currently no information available on hemoplasmas in cattle in the Republic of Korea. The aim of this study was to investigate the presence of hemoplasmas in Korean native cattle and to evaluate the association between hemoplasma infection and anemia.MethodsOne farm was selected, at which blood samples were collected from 104 Korean native cattle [grazing cattle (n = 89) and housed cattle (n = 15)]. Hemoplasmas were detected via polymerase chain reaction analysis and complete blood counts were also performed.ResultsThe overall prevalence of hemoplasmas was 34% (35/104); 20.2% (21/104) for M. wenyonii, 3.8% (4/104) for C. M. haemobos, and 9.6% (10/104) for co-infection. Candidatus Mycoplasma haemobos was detected only in grazing cattle. Of red blood cell (RBC) parameters, C. M. haemobos-infected cattle had lower RBC and hematocrit, and higher mean cell volume than hemoplasma-negative cattle, although none of these differences were statistically significant. This is the first study to report the occurrence of M. wenyonii and C. M. haemobos. Mycoplasma wenyonii is more prevalent than C. M. haemobos in Korean native cattle. The results did not show an association between hemoplasma infection and anemia.ConclusionsConsidering the infection rate of hemoplasmas shown in this study, further studies, such as on the pathogenicity and clinical significance of hemoplasmas are necessary.

Apparent prevalence of hemotropic mycoplasma in dairy calves and replacement heifers on Michigan farms

The bovine hemoplasmas include Mycoplasma wenyonii and Candidatus Mycoplasma hemobos which are increasingly recognized as infecting cattle throughout the world. Infection with hemotropic mycoplasma has been reported to be widespread in mature dairy cows, but little is known about prevalence in calves and heifers. The objective of this study was to investigate the prevalence and dynamics of infection with M. wenyonii and C.M. hemobos infections in calves and replacement heifers on Michigan dairy farms and assess potential associations between infection status and hematological values. The study was designed as a prospective cross-sectional study with a longitudinal component. A convenience sample of 11 farms agreed to participate and were visited twice between March and September 2022. During the first farm visit, researchers collected blood samples from up to 94 animals per farm distributed among newborn and pre-weaned calves (n ≤ 31), weaned calves (n = 21), pre-breeding heifers (n = 21), and pregnant heifers (n = 21). During the first visit, blood samples (n = 174) were also collected from a convenience sample of mature cows to confirm the herd infection status. The same calves and heifers were sampled again about 95 d (±3.0) later. During the first visit, blood samples were collected from 797 calves and replacement heifers, while 675 samples were collected during the second visit due to inability to locate some animals. Detection of M. wenyonii and C. M. hemobos were based on results of real time PCR. The hematocrit was determined using microcentrifugation and the concentration of leukocytes using an automated cell counter. In all herds, most mature cows that were sampled tested positive for infection. The within herd apparent prevalence of hemoplasma in calves and replacement heifers was 100% for both M. wenyonii and C. M. hemobos. The apparent prevalence of hemoplasma in youngstock was associated with age. In calves that were 1 to 6 mo old, the prevalence of infection was 6-8% but sharply increased to 31% by 8 mo of age. In older animals, the prevalence remained high, and was almost 100% in animals greater than 17 mo of age. Based on calves and heifers sampled twice, the cumulative incidence varied widely among herds ranging from 3.7% to 96.0% and increased with age of animals. There was no difference in hematocrit or number of lymphocytes, monocytes, neutrophils, or total leukocytes based on infection status. The number of eosinophils was greater in infected animals. This is the first study to report the prevalence of hemoplasmas in calves and replacement heifers in the US It indicates that young calves can be infected with hemoplasmas, but the rate of infection is low. The likelihood of infection increases as animals age, with a notable rise in the proportion of infected heifers occurring by 8 mo old and the prevalence eventually reaching nearly 100% of infection in older animals. Once infected, heifers appear to remain chronic carriers. Hemoplasma infection alone does not usually lead to the development of clinical signs and most of the animals remain apparently healthy.

High prevalence and genetic diversity of hemoplasmas in bats and bat ectoparasites from China

Hemoplasmas can cause severe hemolytic anemia in humans. To explore the genetic diversity and the potential transmission routes of hemoplasmas among bat population, bats and bat-ectoparasites including bat-flies, bat-mites, and bat-ticks were collected in Eastern and Central China from 2015 to 2021, and tested with PCR for hemoplasmas 16S rRNA gene. Based on 16S rRNA PCR, 18.0% (103/572) adult bats were positive for hemoplasmas, but none of 11 fetuses from hemoplasmas-positive pregnant bats was positive for hemoplasmas. These results indicated that adult bats had a high prevalence of hemoplasma, but vertical transmission of hemoplasmas did not occurr in the bats. Based on the 16S rRNA gene PCR, the minimum infection rate of bat-ectoparasite for hemoplasmas was 4.0% (27/676), suggesting that bat-ectoparasite also had a high prevalence for hemoplasmas. Phylogenetic analysis revealed that bat hemoplasmas from this study clustered into 4 genotypes (I-IV). Genotype I clustered together with hemoplasmas identified in bats from America. Genotype II shared high similarity with a human-pathogenic hemoplasma Candidatus Mycoplasma haemohominis. Genotype III and IV were unique, representing 2 new hemoplasma genotypes. Only genotype I was identified in both bats and all bat-ectoparasites including bat-flies, bat-mites, and bat-ticks. In conclusion, bats and bat-ectoparasites from China harbored abundant genetically diverse hemoplasmas including potential human-pathogenic hemoplasmas, indicating bats and bat-ectoparasites may play important roles in the maintenance and transmission of hemoplasmas in the natural foci.

Prevalence of hemoplasmas and Bartonella species in client-owned cats in Beijing and Shanghai, China.

A year-round molecular epidemiological survey (2017 to 2018) was conducted on three hemoplasmas and two Bartonella species with zoonotic potential in client-owned cats in Beijing and Shanghai. Among 668 specimens, the overall hemoplasma-positive rate was 4.9% (3.4% for Candidatus Mycoplasma haemominutum, 0.9% for Mycoplasma haemofelis and 1.2% for Candidatus Mycoplasma turicensis). The overall Bartonella-positive rate was 8.5% (4.8% for B. henselae and 4.3% for B. clarridgeiae). Age, breed, ectoparasiticide use and stray history, but not city, season and gender, were significantly associated with the positive rates of one or more pathogens. This is also the first report on the prevalence of Candidatus Mycoplasma turicensis in cats in China.

Transfusion transmissible pathogens are prevalent in healthy cats eligible to become blood donors.

This study aims to determine the prevalence of subclinical infectious agents considered core pathogens for worldwide screening in healthy, client-owned, indoor cats eligible to become blood donors in Spain and Portugal. Blood samples of healthy, indoor, domestic cats selected to be potential blood donors were tested for feline leukaemia virus antigens, feline immunodeficiency virus antibodies and polymerase chain reactions for Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum, Candidatus Mycoplasma turicensis, feline leukaemia virus provirus, Leishmania spp. and Bartonella spp. Not all donors were tested for all agents. Overall, 5105 healthy indoor cats were tested and 8.1% (414/5105) had at least one subclinical infectious agent that is transmissible through blood product transfusion. 1.5% (77/5105) were positive for feline leukaemia virus antigens and 2.9% (148/5105) were positive for feline immunodeficiency virus antibodies, therefore they were excluded as donors. The overall prevalence of haemoplasmas in feline leukaemia virus and feline immunodeficiency virus SNAP-negative feline blood donors was 3.7% (181/4880) [1.3% (63/4880) were positive for Mycoplasma haemofelis; 2.3% (112/4880) to Candidatus Mycoplasma haemominutum and 0.12% (6/4880) to Candidatus Mycoplasma turicensis]. The prevalence of feline leukaemia provirus was 5.2% (9/173) and of Bartonella spp. was 0.2% (2/1122). None of the 173 cats were positive for Leishmania spp. The prevalence of many transfusion transmissible pathogens was relatively high in this healthy, client-owned, indoor cats eligible to become blood donors. Performing an extended screening panel that includes at least polymerase chain reactions for Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum, Candidatus Mycoplasma turicensis, feline leukaemia virus provirus, and Bartonella spp., in addition to feline leukaemia virus antigens and feline immunodeficiency virus antibodies, is recommended in pet blood banks from analogous regions.

Polychromophilus melanipherus and haemoplasma infections not associated with clinical signs in southern bent-winged bats (Miniopterus orianae bassanii) and eastern bent-winged bats (Miniopterus orianae oceanensis).

While bats are often viewed as carriers of infectious disease agents, little research has been conducted on the effects these pathogens may have on the bat populations themselves. The southern bent-winged bat (Miniopterus orianae bassanii) is a critically endangered subspecies endemic to south-eastern Australia. Population numbers of this bat have declined over the past 50 years, but the reasons for this are unclear. As part of a larger study to determine if disease could be a contributing factor to this decline, southern bent-winged bats from several locations in Victoria and South Australia were captured and examined for the presence of the blood parasite, Polychromophilus melanipherus, and haemoplasmas (Mycoplasma sp.). Results were compared with those obtained from populations of the more common, partially sympatric, eastern bent-winged bat (Miniopterus orianae oceanensis) from three different locations in Victoria. Both organisms were found in both subspecies (prevalence of P. melanipherus 60% by PCR for southern bent-winged bats compared with 46% for eastern bent-winged bats; prevalence of haemoplasmas 10% for southern bent-winged bats compared with 8% for eastern bent-winged bats), with no association between the probability of infection, body weight, abnormal blood parameters or any other indicators of ill health. However, Victorian southern bent-winged bats had heavier burdens of P. melanipherus than both the South Australian southern bent-winged bats and eastern bent-winged bats. Further investigations are required to determine if these differences are impacting population health.

Novel hemotropic mycoplasmas are widespread and genetically diverse in vampire bats.

Bats (Order: Chiroptera) have been widely studied as reservoir hosts for viruses of concern for human and animal health. However, whether bats are equally competent hosts of non-viral pathogens such as bacteria remains an important open question. Here, we surveyed blood and saliva samples of vampire bats from Peru and Belize for hemotropic Mycoplasma spp. (hemoplasmas), bacteria that can cause inapparent infection or anemia in hosts. 16S rRNA gene amplification of blood showed 67% (150/223) of common vampire bats (Desmodus rotundus) were infected by hemoplasmas. Sequencing of the 16S rRNA gene amplicons revealed three novel genotypes that were phylogenetically related but not identical to hemoplasmas described from other (non-vampire) bat species, rodents, humans, and non-human primates. Hemoplasma prevalence in vampire bats was highest in non-reproductive and young individuals, did not differ by country, and was relatively stable over time (i.e., endemic). Metagenomics from pooled D. rotundus saliva from Peru detected non-hemotropic Mycoplasma species and hemoplasma genotypes phylogenetically similar to those identified in blood, providing indirect evidence for potential direct transmission of hemoplasmas through biting or social contacts. This study demonstrates vampire bats host several novel hemoplasmas and sheds light on risk factors for infection and basic transmission routes. Given the high frequency of direct contacts that arise when vampire bats feed on humans, domestic animals, and wildlife, the potential of these bacteria to be transmitted between species should be investigated in future work.

Mycoplasma ovis infection in goat farms from northeastern Brazil.

Although Mycoplasma ovis (formerly Eperythrozoon ovis) has been described in small ruminants worldwide, data on M. ovis in goats remain scarce. Accordingly, the aims of the present study were to i) determine the prevalence of hemoplasmas in goats, ii) identify the tick species parasitizing the animals, and iii) determine factors associated with infection in five dairy and three beef goat farms from the Paraíba State, northeastern Brazil. Blood samples were obtained from 402 goats. Samples were screened for hemoplasmas using a pan-hemoplasma PCR. The positive samples were confirmed by sequencing. An epidemiological questionnaire was given to each farm owner addressing age, gender, and presence of ticks. A total of 158/402 (39.3%) goats were positive for M. ovis by PCR. Sequencing of PCR positive samples has shown ≥99% identity with multiple M. ovis 16S rDNA sequences deposited in GenBank, including M. ovis isolates from humans. Dairy (OR=2.15; 95% CI: 1.40-3.32%; P=0.0004) and anemic goats (OR=2.33; 95% CI: 1.51-3.71%; P=0.0001) were more likely to be infected than beef and non-anemic animals, respectively. Amblyomma parvum (49/52, 94.23%) and Rhipicephalus microplus (3/52, 5.77%) were the tick species found parasitizing the animals, with no significant association between the presence of ticks and infection by M. ovis (P=0.1164). This is the first reportedly molecular detection of M. ovis infection in goats from South America. In conclusion, M. ovis is highly prevalent in goats from northeastern Brazil, mainly in dairy animals.

Molecular characterization of hemotropic mycoplasmas (Mycoplasma ovis and \u2018Candidatus Mycoplasma haemovis\u2019) in sheep and goats in China

BackgroundHemotropic mycoplasmas (hemoplasmas) are emerging zoonotic pathogens with a worldwide distribution that can cause mild to severe hemolytic anemia, icterus, ill-thrift, infertility, and poor weight gain. However, understanding of the molecular epidemiology of hemoplasmas (Mycoplasma ovis and ‘Candidatus Mycoplasma haemovis’) is limited in sheep and goats, and the hemoplasma strain/species/variant ‘Candidatus M. haemovis’ was poorly studied throughout the world and had never been detected in China until now. Thus, the aim of the present study was to determine the molecular prevalence of hemoplasmas, including M. ovis and ‘Candidatus M. haemovis’ in sheep and goats from seven provinces and one autonomous region of China.MethodsA total of 1364 blood samples were collected from sheep and goats in seven provinces and one autonomous region of China. All blood samples were tested for hemoplasmas (M. ovis and ‘Candidatus M. haemovis’) by nested PCR amplification based on 16S rRNA gene. Positive specimens underwent nucleotide sequencing and phylogenetic analysis.ResultsOverall, 610 specimens (44.7%, 610/1364) were shown to be hemoplasmas (M. ovis and ‘Candidatus M. haemovis’) -positive by nested PCR amplification based on 16S rRNA gene. The prevalence in goats was 44.1% (379/860), and 45.8% (231/504) in sheep, while that in grazing small ruminants was 54.4% (396/728) and 33.6% (214/636) in house feeding small ruminants. Sequencing of the nearly complete 16S rRNA gene was successful for the 103 randomly selected positive specimens from different farms in different sampling sites of China. Among them, analysis of the 16S rRNA gene sequences identified M. ovis (n = 56) and ‘Candidatus M. haemovis’ (n = 47). Two (KU983740 and KU983746) of the four novel genotypes obtained in this study were closely related to M. ovis, while the other two genotypes (KU983748 and KU983749) had high identity with ‘Candidatus M. haemovis’. Remarkably, the genotype (KU983740) of M. ovis in sheep and goats in this study fell in a clade with two human hemoplasmas from USA (KF313922 and GU230144) and shared 99.8%–99.9% with them.ConclusionsIn this study, ‘Candidatus M. haemovis’ was first detected in Chinese sheep and goats and hemoplasmas (M. ovis and ‘Candidatus M. haemovis’) are highly prevalent, and widely distributed in China.

Risk factors of different hemoplasma species infections in cats.

BackgroundHemoplasma species (spp.) commonly cause infections in cats worldwide. However, data on risk factors for infections are limited. The aim of this study was to determine the prevalence of hemoplasma spp. infections in cats in Southern Germany and to assess risk factors associated with infection.ResultsDNA was extracted from blood samples of 479 cats presented to different veterinary hospitals for various reasons. DNA of feline hemoplasmas was amplified by use of a previously reported PCR assay. Direct sequencing was used to confirm all purified amplicons and compared to hemoplasma sequences reported in GenBank. Results were evaluated in relation to the age, sex, housing conditions, feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) status of the cats.The overall hemoplasma prevalence rate was 9.4% (45/479; 95% CI: 7.08–12.36). ‘Candidatus Mycoplasma (M.) haemominutum’ (Mhm) DNA was amplified from 42 samples, M. haemofelis from 2, and M. haemocanis from 1 sample. There was a significantly higher risk of hemoplasma infection in cats from multi-cat households, in outdoor cats, as well as in cats with FIVinfection and in cats with abortive FeLV infection, but not in cats with progressive or regressive FeLV infection.ConclusionsMhm infection is common in cats in Southern Germany. Higher prevalence in multi-cat households and associations with FeLV infection likely reflect the potential for direct transmission amongst cats. Outdoor access, male gender, and FIV infection are additional risk factors that might relate to aggressive interactions and exposure to vectors.

MOLECULAR INVESTIGATION OF HEMOTROPIC MYCOPLASMAS IN HUMAN BEINGS, DOGS AND HORSES IN A RURAL SETTLEMENT IN SOUTHERN BRAZIL.

SUMMARYThe aims of this study were to determine the prevalence of hemoplasmas in a rural Brazilian settlement's population of human beings, their dogs and horses, highly exposed to tick bites; to identify the tick species parasitizing dogs and horses, and analyze factors associated with their infection. Blood samples from 132 dogs, 16 horses and 100 humans were screened using a pan-hemoplasma SYBR green real-time PCR assay followed by a species-specific TaqMan real-time PCR. A total of 59/132 (44.7%) dog samples were positive for hemoplasmas (21 Mycoplasma haemocanisalone, 12 ' Candidatus Mycoplasma haematoparvum' alone and 21 both). Only 1/100 (1.0%) human sample was positive by qPCR SYBR green, with no successful amplification of 16S rRNA or 23 rRNA genes despite multiple attempts. All horse samples were negative. Dogs >1 year of age were more likely to be positive for hemoplasmas ( p= 0.0014). In conclusion, although canine hemoplasma infection was highly prevalent, cross-species hemoplasma transmission was not observed, and therefore may not frequently occur despite overexposure of agents and vectors.

Hemoplasma prevalence and hematological abnormalities associated with infection in three different cat populations from Southern Brazil.

Three hemoplasma species are recognized in domestic cats: Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis'. We report the prevalence and hematological abnormalities of hemoplasma infection in 369 domestic cats from three different populations (blood donors, hospitalized cats and shelter cats) from Southern Brazil. Complete blood counts were performed at the time of blood collection, and DNA was extracted and tested by conventional PCR for each hemoplasma species. A total of 79 samples (21.40%) were positive for at least one species. The most prevalent hemoplasma was 'Candidatus Mycoplasma haemominutum', with 50/369 (13.55%) positive cats, followed by 'Candidatus Mycoplasma turicensis', 10/369 (2.71%), and Mycoplasma haemofelis, 8/369 (2.16%). Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' coinfection was observed in 4/369 (1.08%), whereas 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis' in 5/369 (1.35%). Three cats (0.81%) were infected with all three hemoplasmas. There was no association between infection and the different populations. Anemia was associated with Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum', but not with 'Candidatus Mycoplasma turicensis'. Male cats and cats with outdoor access were more likely to be infected. Although 'Candidatus Mycoplasma haemominutum' is believed to cause minimal or no hematological alterations, the infected cats studied herein were more likely to be anemic.

First evidence of hemoplasma infection in free-ranging Namibian cheetahs (Acinonyx jubatus)

Infections with feline hemotropic mycoplasmas (hemoplasmas) have been documented in domestic cats and free-ranging feline species with high prevalences in Iberian lynxes (Lynx pardinus), Eurasian lynxes (Lynx lynx), European wildcats (Felis silvestris silvestris), African lions (Panthera leo) in Tanzania and domestic cats in South Africa. The prevalence of hemoplasmas has not yet been investigated in free-ranging felids in southern Africa. In this study we screened 73 blood samples from 61 cheetahs in central Namibia for the presence of hemoplasmas using quantitative real-time PCR. One of the cheetahs tested PCR-positive. Phylogenetic analysis based on partial sequencing of the 16S rRNA and RNAse P genes revealed that the isolate belongs to the Mycoplasma haemofelis/haemocanis group. This is the first molecular evidence of a hemoplasma infection in a free-ranging cheetah.

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’...

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.

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.

Differential Detection of Hemotropic Mycoplasma Species in Cattle by Melting Curve Analysis of PCR Products

We developed a real-time PCR procedure followed by melting curve analysis using the green fluorescence dye SYBR Green I for rapid detection and differentiation of hemplasmas in cattle. Analysis of the melting temperature (Tm) of the PCR products allowed for differentiation of the 2 bovine hemoplasmas, Mycoplasma wenyonii and a provisional species, ;Candidatus Mycoplasma haemobos' (a synonym of ;Candidatus M. haemobovis'). The Tm (mean +/- S.E.) of the PCR products from the bovine hemoplasmas were 86.98 +/- 0.12 degrees C for M. wenyonii and 82.04 +/- 0.27 degrees C and 86.98 +/- 0.12 degrees C, respectively; [corrected] Candidatus M. haemobos' in the melting experiments. The protocol described in the present study can decrease the time to results by simultaneous detection and differentiation of the two hemoplasmas in cattle. By using this protocol, we examined hemoplasma prevalence in 109 cattle in Miyagi Prefecture and found that 67 (61.5%) were infected with M. wenyonii, 25 (22.9%) were infected with ;Candidatus M. haemobos' and 14 (12.8%) were infected with both.

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.