'Candidatus Mycoplasma haematohydrochoerus', a novel hemoplasma species in capybaras (Hydrochoerus hydrochaeris) from Brazil.

Three different species of hemoplasmas have been described in rodents, Mycoplasma coccoides, 'Candidatus Mycoplasma haemomuris' and 'Candidatus Mycoplasma haemosphiggurus'. Additionally, potentially novel hemoplasma species have been detected in wild rodents from Brazil, including capybaras (Hydrochoerus hydrochaeris). Capybaras are the largest rodent in the world and are well adapted to live within close proximity to humans, which increases the risk to spread of zoonotic pathogens. Herein, we investigate the occurrence and genetic diversity of hemoplasmas infecting free-ranging capybaras from southern Brazil. Blood samples and ticks from 17 capybaras were collected. Packed cell volume and total plasma protein were measured, DNA was extracted, and further screened by species-specific and pan-hemoplasma PCR assays targeting the 16S rRNA gene of hemoplasmas. Sixteen out of 17 (94.12%; 95% CI: 73.02-98.95%) were anemic. Only one young female was hypoproteinemic. All capybaras were infested by adults and nymphs of Amblyomma dubitatum ticks. Using the PCR assay targeting the 16S rRNA gene of M. coccoides, 13/17 (76.47%; 95% CI: 52.74-90.44%) capybaras were positive for hemoplasmas. When DNA samples were tested by the pan-hemoplasma PCR, 16/17 (94.12%; 95% CI: 73.02-98.95%) animals were positive. One out of 11 (9.09%) adult ticks salivary glands tested positive for hemoplasma by the pan-hemoplasma PCR assay. Sequencing and phylogenetic analysis of the 16S and 23S rRNA gene fragments confirmed that animals were infected by a novel hemotropic Mycoplasma sp. previously reported in capybaras from Brazil. Additionally, sequencing and phylogenetic analysis of the 23S rRNA gene from three hemoplasma-positive capybaras samples from a previous study performed in midwestern Brazil also confirm our findings. Based on phylogenetic and Neighbor-Net network analysis of the 16S rRNA and 23S rRNA genes, the name 'Candidatus Mycoplasma haematohydrochoerus' is proposed for this novel organism.

House finches with high coccidia burdens experience more severe experimental Mycoplasma gallisepticum infections.

Parasites co-infecting hosts can interact directly and indirectly to affect parasite growth and disease manifestation. We examined potential interactions between two common parasites of house finches: the bacterium Mycoplasma gallisepticum that causes conjunctivitis and the intestinal coccidian parasite Isospora sp. We quantified coccidia burdens prior to and following experimental infection with M. gallisepticum, exploiting the birds' range of natural coccidia burdens. Birds with greater baseline coccidia burdens developed higher M. gallisepticum loads and longer lasting conjunctivitis following inoculation. However, experimental inoculation with M. gallisepticum did not appear to alter coccidia shedding. Our study suggests that differences in immunocompetence or condition may predispose some finches to more severe infections with both pathogens.

Occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens

Mycoplasmaspp. and Ureaplasmaspp. belong tohumans'genitourinary microbiota and sometimesare associated with infections of the genitourinarytract. The aim of this study was to evaluate the occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens from patients of the 15thRegional de Saúde of ParanáState, Brazil, and to correlate the results with clinical and laboratory data.A retrospective cross-sectional study was conducted,based on the analysis of results of vaginal, endocervical, urine andurethral culture for mycoplasmas from patients attended in areference laboratory, from January 2009 to December 2016. We evaluated 2,475 results of culture for mycoplasmas. A total of 50.8% patients were positive for mycoplasmas. Of these, 76.8%had positive culture exclusively for Ureaplasmaspp. and 4.7% for Mycoplasmahominis. Both microorganisms were isolated in the microbiology culture of 18.5% of patients. Among the positive culture, 81.4% had significant concentrations.Bacterialvaginosis was the most common alteration observed in association with mycoplasmas.Thehigh positivity of cultures for mycoplasmas, especially Ureaplasmaspp. found in our study, highlightthe presence of these microorganisms in many of the genital tract disorders that can be sexually transmitted and, consequently, should not be neglected.

Neglected intravascular pathogens, Babesia vulpes and haemotropic Mycoplasma spp. in European red fox (Vulpes vulpes) population.

Wild animals, especially canids, are important reservoirs of vector-borne pathogens, that are transmitted by the ticks and other bloodsucking arthropods. In total, 300 red foxes (Vulpes vulpes), shot by the hunters in eastern and northern Slovakia, were screened for the presence of vector-borne pathogens by PCR-based methods Blood samples were obtained from nine red foxes and tissue samples originated from 291 animals (the liver tissue samples from 49 foxes and spleen samples from 242 red foxes). Babesia vulpes and haemotropic Mycoplasma species were identified by amplification and sequencing of 18S rRNA and 16S rRNA gene fragments, respectively. Overall, the presence of these pathogens was recorded in 12.3% of screened DNA samples. Altogether 9.7% (29/300) of investigated foxes carried DNA of Babesia spp. In total, 12 out of 29 Babesia spp. PCR - positive amplicons were further sequenced and identified as B. vulpes (41.4%; 12/29), remaining 17 samples are referred as Babesia sp. (58.6%; 17/29). Overall prevalence of B. vulpes reached 4.0% (n=300). Thirteen (4.3%) samples tested positive for distinct Mycoplasma species. To the best of our knowledge, this study brings the first information on B. vulpes infection in red foxes in Slovakia, and the first data on the prevalence and diversity of haemotropic Mycoplasma spp. in European red fox population. Moreover, co-infections with B. vulpes and Mycoplasma spp. were confirmed in 1.7% of tested DNA samples. The relatively high rates of blood pathogen' prevalence and species diversity in wild foxes indicate the role of the fox population in the maintenance of the parasites in sylvatic cycles and strengthen the assumption that foxes play an important role in spreading of infectious microorganisms within and outside the natural foci.

Canine tick-borne pathogens in Cyprus and a unique canine case of multiple co-infections.

Canine tick-borne pathogens such as Ehrlichia canis and Hepatozoon canis are widespread in the Mediterranean basin but have never been reported or investigated in Cyprus. We describe herein the presence of canine tick-borne pathogens in three dogs with clinical signs compatible with vector-borne diseases from Paphos area of Cyprus. Molecular and phylogenetic analysis revealed the presence of E. canis, Anaplasma platys, H. canis, Babesia vogeli and Mycoplasma haemocanis in Cyprus. One dog co-infected with E. canis, H. canis, B. vogeli and M. haemocanis is, to the best of our knowledge, the first report of this multiple co-infection in dogs. The tick-borne pathogens reported in the current study should be considered in the differential diagnoses in dogs exposed to ticks in Cyprus.

Hematological changes associated with hemoplasma infection in cats in Rio de Janeiro, Brazil.

This study aimed to detect Mycoplasma spp. in naturally infected cats from Rio de Janeiro and to evaluate hematological abnormalities and factors associated with this infection. Out of the 197 cats sampled, 11.2% presented structures compatible with hemoplasma organisms on blood smears. In contrast, 22.8% were positive for Mycoplasma spp. by means of 16S rRNA gene real-time polymerase chain reaction, which reflects the weak concordance between techniques. The infection rates, by means of 16S rRNA gene conventional polymerase chain reaction, was 4.6%, 4.6% and 11.7% for Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma turicensis' (CMt) and 'Candidatus Mycoplasma haemominutum' (CMhm), respectively. Mhf and CMhm infections are more frequent in the summer (p>0.05). Presence of anemia (p < 0.02), lymphocytosis (p < 0.03), thrombocytopenia (p < 0.04) and activated monocytes (p < 0.04) was associated with Mhf infection. No hematological abnormality was associated with CMt or CMhm infection. Male cats were more prone to be infected by Mhf or CMhm (p < 0.01). Adult cats had more chance to be infected by CMhm. Three hemoplasma species occur in the metropolitan region of Rio de Janeiro and Mhf seems to be the most pathogenic of them. Anemia is the most important hematological abnormality.

Hematological changes associated with hemoplasma infection in cats in Rio de Janeiro, Brazil / Alterações hematológicas associadas à infecção por hemoplasmas em gatos do Rio de Janeiro, Brasil

Abstract This study aimed to detect Mycoplasma spp. in naturally infected cats from Rio de Janeiro and to evaluate hematological abnormalities and factors associated with this infection. Out of the 197 cats sampled, 11.2% presented structures compatible with hemoplasma organisms on blood smears. In contrast, 22.8% were positive for Mycoplasma spp. by means of 16S rRNA gene real-time polymerase chain reaction, which reflects the weak concordance between techniques. The infection rates, by means of 16S rRNA gene conventional polymerase chain reaction, was 4.6%, 4.6% and 11.7% for Mycoplasma haemofelis (Mhf), ‘Candidatus Mycoplasma turicensis’ (CMt) and ‘Candidatus Mycoplasma haemominutum’ (CMhm), respectively. Mhf and CMhm infections are more frequent in the summer (p>0.05). Presence of anemia (p < 0.02), lymphocytosis (p < 0.03), thrombocytopenia (p < 0.04) and activated monocytes (p < 0.04) was associated with Mhf infection. No hematological abnormality was associated with CMt or CMhm infection. Male cats were more prone to be infected by Mhf or CMhm (p < 0.01). Adult cats had more chance to be infected by CMhm. Three hemoplasma species occur in the metropolitan region of Rio de Janeiro and Mhf seems to be the most pathogenic of them. Anemia is the most important hematological abnormality.

Resumo Este estudo teve por objetivo detectar Mycoplasma spp. em gatos naturalmente infectados do Rio de Janeiro e avaliar as alterações hematológicas e fatores associados à infecção. Dos 197 gatos amostrados, 11,2% apresentaram estruturas compatíveis com hemoplasmas em esfregaços de sangue. Em contraste, 22,8% foram positivas para Mycoplasma spp. por meio da reação em cadeia da polimerase em tempo real (qPCR), baseado no gene 16S rRNA, o que reflete a fraca concordância entre as técnicas. As taxas de infecção, por meio da reação em cadeia da polimerase convencional baseada no gene 16S rRNA, foi de 4,6%, 4,6% e 11,7% para Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma turicensis' (CMt) e 'Candidatus Mycoplasma haemominutum' (CMhm), respectivamente. Infecção por Mhf e CMhm foram mais frequentes no verão (p> 0,05). Anemia (p<0,02), linfocitose (p<0,03), trombocitopenia (p<0,04), e presença de monócitos ativados (p<0,04) foram associados à infecção por Mhf. Nenhuma alteração hematológica foi associada à infecção por CMt ou CMhm. Gatos machos estão mais propensos à infecção por Mhf ou CMhm (p<0,01). Gatos adultos têm maiores chances de se infectarem por CMhm. Há ocorrência de três espécies de hemoplasmas na Região Metropolitana do Rio de Janeiro e Mhf parece ser o mais patogênico, tendo a anemia como principal alteração hematológica.

Occurrence of Mycoplasma haemocanis in dogs infested by ticks in Campo Grande, Mato Grosso do Sul, Brazil.

Hemotropic mycoplasmas in dogs, such as Mycoplasma haemocanis, have been described worldwide. Recently, these pathogens have been reported to be causative agent of zoonosis. It is known that its transmission may occur through the action of blood-sucking arthropods (e.g. ticks or fleas), through blood transfusion, contaminated fomites and/or transplacentally. In Brazil, M. haemocanis is present in practically all regions and the tick Rhipicephalus sanguineus sensu lato is suspected the main vector. In the municipality of Campo Grande, state of Mato Grosso do Sul, there is little information about infection of dogs by M. haemocanis, or on the main epidemiological features associated with it. Thus, the aim of the present study was to determine the occurrence of M. haemocanis among dogs infested by ticks and to assess possible associations with some epidemiological factors. The polymerase chain reaction (PCR) and DNA sequencing were used to analyze dog blood samples (n = 94). DNA from M. haemocanis was detected in four samples. No significant associations were observed with any epidemiological parameter analyzed here. However, the results from this study confirm that this pathogen is circulating in this region and should be considered in the differential diagnosis of diseases among anemic dogs.

Occurrence of Mycoplasma haemocanis in dogs infested by ticks in Campo Grande, Mato Grosso do Sul, Brazil / Ocorrência de Mycoplasma haemocanis em cães infestados por carrapatos em Campo Grande, Mato Grosso do Sul, Brasil

Abstract Hemotropic mycoplasmas in dogs, such as Mycoplasma haemocanis, have been described worldwide. Recently, these pathogens have been reported to be causative agent of zoonosis. It is known that its transmission may occur through the action of blood-sucking arthropods (e.g. ticks or fleas), through blood transfusion, contaminated fomites and/or transplacentally. In Brazil, M. haemocanis is present in practically all regions and the tick Rhipicephalus sanguineus sensu lato is suspected the main vector. In the municipality of Campo Grande, state of Mato Grosso do Sul, there is little information about infection of dogs by M. haemocanis, or on the main epidemiological features associated with it. Thus, the aim of the present study was to determine the occurrence of M. haemocanis among dogs infested by ticks and to assess possible associations with some epidemiological factors. The polymerase chain reaction (PCR) and DNA sequencing were used to analyze dog blood samples (n = 94). DNA from M. haemocanis was detected in four samples. No significant associations were observed with any epidemiological parameter analyzed here. However, the results from this study confirm that this pathogen is circulating in this region and should be considered in the differential diagnosis of diseases among anemic dogs.

Resumo Micoplasmas hemotrópicos de cães, como Mycoplasma haemocanis, já foram descritos em todo o mundo. Recentemente, esses patógenos têm sido apontados como causadores de zoonoses. É sabido que a transmissão pode ocorrer devido à ação de artrópodes sugadores de sangue (carrapatos, pulgas), transfusão sanguínea e/ou fômites contaminados e por via transplacentária. No Brasil, Mycoplasma haemocanis está presente em praticamente todas as regiões, e o carrapato Rhipicephalus sanguineus sensu lato é suspeito como principal vetor. No município de Campo Grande, Estado de Mato Grosso do Sul, Brasil não existem muitas informações acerca de infecções de cães por M. haemocanis, assim como quais são os principais aspectos epidemiológicos associados a este patógeno. Assim, o objetivo, no presente estudo, foi determinar a ocorrência de M. haemocanis em cães infestados por carrapatos e analisar possíveis associações com alguns fatores epidemiológicos. A Reação em Cadeia da Polimerase (PCR) e o sequenciamento de DNA foram utilizados para analisar amostras de sangue de cães (n = 94). DNA de M. haemocanis foi identificado em quatro amostras. Não foram observadas associações significativas com qualquer parâmetro epidemiológico analisado. No entanto, os resultados deste estudo confirmam que esse patógeno está circulando na região e deve ser considerado no diagnóstico diferencial de causas de anemia em cães.

Development and evaluation of a multi-locus sequence typing scheme for Mycoplasma synoviae.

Reproducible molecular Mycoplasma synoviae typing techniques with sufficient discriminatory power may help to expand knowledge on its epidemiology and contribute to the improvement of control and eradication programmes of this mycoplasma species. The present study describes the development and validation of a novel multi-locus sequence typing (MLST) scheme for M. synoviae. Thirteen M. synoviae isolates originating from different poultry categories, farms and lesions, were subjected to whole genome sequencing. Their sequences were compared to that of M. synoviae reference strain MS53. A high number of single nucleotide polymorphisms (SNPs) indicating considerable genetic diversity were identified. SNPs were present in over 40 putative target genes for MLST of which five target genes were selected (nanA, uvrA, lepA, ruvB and ugpA) for the MLST scheme. This scheme was evaluated analysing 209 M. synoviae samples from different countries, categories of poultry, farms and lesions. Eleven clonal clusters and 76 different sequence types (STs) were obtained. Clustering occurred following geographical origin, supporting the hypothesis of regional population evolution. M. synoviae samples obtained from epidemiologically linked outbreaks often harboured the same ST. In contrast, multiple M. synoviae lineages were found in samples originating from swollen joints or oviducts from hens that produce eggs with eggshell apex abnormalities indicating that further research is needed to identify the genetic factors of M. synoviae that may explain its variations in tissue tropism and disease inducing potential. Furthermore, MLST proved to have a higher discriminatory power compared to variable lipoprotein and haemagglutinin A typing, which generated 50 different genotypes on the same database.

Investigation of bovine haemoplasmas and their association with anaemia in New Zealand cattle.

CASE HISTORY AND CLINICAL FINDINGS: A dairy cow, from a herd in the Waikato region of New Zealand, was reported with regenerative anaemia on 12 September 2014. Testing of blood from the animal using PCR assays for Theileria orientalis produced a negative result for both Chitose and Ikeda types. LABORATORY FINDINGS: Using PCR and DNA sequencing, blood from the cow was positive for Candidatus Mycoplasma haemobos. Further testing of another 12 animals from the case herd, 27 days after the affected cow was first reported, showed 11 animals were positive for Candidatus M. haemobos or Mycoplasma wenyonii in the PCR. None of these cattle were clinically anaemic or positive for T. orientalis Ikeda type using PCR. A convenience sample of 47 blood samples from cattle throughout New Zealand, submitted to the Investigation and Diagnostic Centre (Ministry for Primary Industries) for surveillance testing for T. orientalis Ikeda, was selected for further testing for bovine haemoplasmas. Of these samples, 6/47 (13%) and 13/47(28%) were positive for M. wenyonii and Candidatus M. haemobos, respectively. There was no difference in the proportion of samples positive for the bovine haemaplasmas between cattle with anaemia that were negative for T. orientalis (6/20, 33%), or without anaemia or T. orientalis (10/18, 56%), or from cattle herds experiencing anaemia and infection with T. orientalis Ikeda type (3/9, 33%). DIAGNOSIS: Bovine haemoplasmosis. CLINICAL RELEVANCE: The presence of bovine haemoplasmas in blood does not establish causality for anaemia in cattle. Diagnosis of anaemia associated with haemoplasmosis would require exclusion of other causes of regenerative anaemia and an association of the agent with anaemia in affected cattle herds. The data collected in this study did not provide evidence that bovine haemoplasmas were associated with a large number of outbreaks of anaemia in cattle in New Zealand.

Prevalence of Trichomonas vaginalis by PCR in men attending a primary care urology clinic in South Korea.

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

Prevalence of Trichomonas vaginalis by PCR in Men Attending a Primary Care Urology Clinic in South Korea

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a Brazilian domestic cat (Felis catus).

This article describes the first detection of Cytauxzoon felis, using molecular techniques, in a naturally infected domestic cat from Brazil, South America. Coinfection with 'Candidatus Mycoplasma haemominutum' was also found. The molecular identification of the piroplasmid species was performed by Polymerase Chain Reaction (PCR) and sequencing analysis. A 284 pb fragment of the gene encoding the 18S ribosomal RNA region was amplified and showed 99% identity with other C. felis strains from North America. In addition, PCR-RFLP (restriction fragment length polymorphism) analysis, which amplifies a 595 bp fragment of the gene encoding 16S ribosomal RNA of some bacterial species, identified the co-infecting species as 'Candidatus M. haemominutum'.

Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a brazilian domestic cat (Felis catus) / Co-infecção por Cytauxzoon felis e 'Candidatus Mycoplasma haemominutum' em um gato doméstico (Felis catus) no Brasil

This article describes the first detection of Cytauxzoon felis, using molecular techniques, in a naturally infected domestic cat from Brazil, South America. Coinfection with 'Candidatus Mycoplasma haemominutum' was also found. The molecular identification of the piroplasmid species was performed by Polymerase Chain Reaction (PCR) and sequencing analysis. A 284 pb fragment of the gene encoding the 18S ribosomal RNA region was amplified and showed 99% identity with other C. felis strains from North America. In addition, PCR-RFLP (restriction fragment length polymorphism) analysis, which amplifies a 595 bp fragment of the gene encoding 16S ribosomal RNA of some bacterial species, identified the co-infecting species as 'Candidatus M. haemominutum'.

Este artigo descreve a primeira detecção de Cytauxzoon felis em um gato doméstico naturalmente infectado no Brasil, América do Sul, através de técnicas moleculares. Também foi encontrada co-infecção com 'Candidatus Mycoplasma haemominutum'. A detecção molecular da espécie do piroplasmídeo foi realizada através da reação em cadeia pela polimerase (PCR) e sequenciamento. Um fragmento de 284 pb do gene codificador da região 18S do RNA ribossomal do parasito foi sequenciada e mostrou 99% de identidade com outros isolados de C. felis da América do Norte. Ademais, através da análise por meio de PCR-RFLP (Polimorfismo no comprimento de fragmentos de restrição), que amplifica um fragmento de 595 pb do gene codificador da porção 16 do RNA ribossomal de algumas espécies de bactérias, concluiu-se que a espécie com-infectante era 'Candidatus M. haemominutum'.

Use of real-time quantitative PCR to document successful treatment of Mycoplasma haemocanis infection with doxycycline in a dog.

An 8-year-old Jack Russell Terrier was diagnosed with hemolytic anemia caused by hemoplasmosis 4 years following splenectomy. Quantitative real-time PCR (qPCR) analysis was used initially to confirm infection with Mycoplasma haemocanis and subsequently to monitor and direct medical therapy. Doxycycline was administered beyond resolution of clinical signs until hemoplasma DNA could no longer be detected by qPCR. The dog remained clinically healthy and hemoplasma-negative 8 months following cessation of therapy. Canine hemoplasmosis should remain as a differential diagnosis for hemolytic anemia in dogs, particularly those that are splenectomized or immunocompromised, even in geographic regions where prevalence of infection is low. Prolonged doxycycline administration has been shown by qPCR to lead to sustained absence of detectable infection and should be considered as a first line treatment for canine hemoplasmosis.

[Mycoplasma hominis symbiosis and Trichomonas vaginalis metronidazole resistance].

OBJECTIVE: To investigate the relation of Mycoplasma hominis symbiosis and the resistance of Trichomonas vaginalis to metronidazole. METHODS: From November 2010 to July 2011, 160 isolates of T. vaginalis were collected from the genital tract secretion of gynecological out-patients at the Sichuan Provincial Hospital for Women and Children. The minimum lethal concentration (MLC) to metronidazole of these isolates was determined by an in vitro sensitivity assay with different concentration gradients of metronidazole (from 1 to 1 024 microg/ml), and M. hominis DNA in T. vaginalis was detected by polymerase chain reaction (PCR) technique with specific 16S rRNA primers. After clearance of M. hominis from the parasites by 32 microg/ml doxycycline, MLC was determined and compared with that before clearance. RESULTS: MLC of metronidazole in T. vaginalis ranged from 1 to 256 microg/ml, with 61.3% isolates (98/160) ranging from 1 to 8 microg/ml, 26.3% isolates (42/160) ranging from 16 to 32 microg/ml, and 12.5% isolates (20/160) ranging from 64 to 256 microg/ml. 61 isolates were PCR positive for M. hominis DNA in the 160 isolates of T. vaginalis. The M. hominis DNA positive rate was significantly higher in the T. vaginalis isolates with higher MLC than those isolates with lower MLC (P<0.01). However, when M. hominis was cleared by doxycycline from 8 isolates among the 61 ones, no change was observed in sensitivity of the isolates to metronidazole. CONCLUSION: M. hominis symbiosis might be associated with the metronidazole-resistance of T. vaginalis. However, it needs direct evidence.

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.

Rapid detection of coinfections by Trichomonas vaginalis, Mycoplasma hominis, and Ureaplasma urealyticum by a new multiplex polymerase chain reaction.

We developed a multiplex polymerase chain reaction (M-PCR) assay to simultaneously detect Trichomonas vaginalis, Mycoplasma hominis, and Ureaplasma urealyticum. The test is extremely specific and has a sensitivity of 10 cells for T. vaginalis and U. urealyticum and of 1 cell for M. hominis. The technique was validated on vaginal swabs from 240 women presenting symptoms of vaginitis, and results were compared with data obtained using microscopic and culture techniques on the same patients. The M-PCR revealed to be greatly more sensitive and specific than traditional techniques. It has been well demonstrated, in vitro, that T. vaginalis can establish a symbiosis with M. hominis; our data confirm in vivo this strict association: in fact, M. hominis has been detected in 78.6% of all samples positive for T. vaginalis, as compared to only 4.8% of women without trichomoniasis. The species specificity of this association has been confirmed by the absence of any significant correlation between T. vaginalis and U. urealyticum.

Mycoplasma gallisepticum infection in house finches (Carpodacus mexicanus) affects mosquito blood feeding patterns.

Disease-induced lethargy can diminish host capacity to repel or kill biting mosquitoes. We exposed house finches (Carpodacus mexicanus) to mosquitoes (Culex pipiens pipiens), repeated the experiment after inoculating finches with Mycoplasma gallisepticum, and then repeated the experiment with the same birds after curing their infections. We videotaped avian behaviors before and during mosquito exposure, identifying hosts through blood meal DNA fingerprinting. Results revealed heterogeneity in mosquito preference regardless of infection. Mosquitoes choosing between two healthy finches were more likely to feed upon the same individual bird consistently. When one bird was sick, mosquitoes exhibited no preference. Sick birds made fewer total defensive behaviors than healthy birds, but only foot stomps were associated with reduced mosquito feeding success. Our results suggest that Mycoplasma and other avian infections that alter bird defensive behavior may influence mosquito feeding patterns and transmission of arthropod-borne pathogens such as West Nile virus.