RESUMO
Our study was designed to investigate the original spectrum of feline respiratory tract infection and to provide a scientific basis for the clinical diagnosis and treatment of feline respiratory infections and for precise prevention and control measures. A total of 400 cats with upper respiratory tract infections from animal hospitals in 12 provinces in China were examined from November 2022 to October 2023 to investigate the epidemiology of feline calicivirus (FCV), feline herpes virus type 1 (FHV-1), influenza A virus (IAV), Mycoplasma felis, Chlamydia felis, and Bordetella bronchiseptica through loop-mediated isothermal amplification (LAMP) with microfluidic chip detection. The results showed that 396 of the 400 samples tested were positive for at least one of these pathogens, with an overall detection rate of 99.00%. The detection rates were as follows: FCV, 36.00% (144/400); M. felis, 34.00% (136/400); FHV-1, 21.50% (86/400); C. felis, 15.75% (63/400); B. b, 13.00% (52/400); IAV, 4.50% (18/400). There were no statistically significant differences in the detection rates of respiratory pathogens between different sexes, ages, seasons, breeds, or regions (P > 0.05). There were 88 mixed infections, giving a total mixed infection rate of 22.00% (88/400). It is worth noting that the detection rate of FCV at different ages and of FHV-1 in different sexes showed significant differences (P < 0.05). The highest rate of FCV infection was found in animals that were 1 to 2 years old, and the rate of FHV-1 infection in male cats was higher than that in female cats. The results showed that the spectrum of feline respiratory pathogens is complex, with diverse epidemiological characteristics and mixed infections, and some differences among different respiratory pathogens were found with regard to the sex, age, and breed of the cat. Studies should be continued to provide a scientific basis for precise prevention and control of feline respiratory diseases.
Assuntos
Doenças do Gato , Técnicas de Amplificação de Ácido Nucleico , Infecções Respiratórias , Animais , Gatos , Infecções Respiratórias/veterinária , Infecções Respiratórias/virologia , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/microbiologia , Infecções Respiratórias/diagnóstico , Doenças do Gato/virologia , Doenças do Gato/epidemiologia , Doenças do Gato/microbiologia , Feminino , Masculino , China/epidemiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Calicivirus Felino/isolamento & purificação , Calicivirus Felino/genética , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza A/genética , Vírus da Influenza A/classificação , Chlamydia/genética , Chlamydia/isolamento & purificação , Chlamydia/classificação , Bordetella bronchiseptica/isolamento & purificação , Bordetella bronchiseptica/genética , Mycoplasma/isolamento & purificação , Mycoplasma/genética , Mycoplasma/classificação , Técnicas de Diagnóstico Molecular/métodos , Varicellovirus/genética , Varicellovirus/isolamento & purificação , Varicellovirus/classificação , Sistema Respiratório/virologia , Sistema Respiratório/microbiologiaRESUMO
BACKGROUND: Cats are hosts and reservoirs for many haemopathogens such as piroplasms, Rickettsia, hemotropic Mycoplasma, Bartonella, Ehrlichia, and Anaplasma, which are transmitted by various vector arthropods and some of which have a zoonotic concern. Although it is noteworthy that the rate of ownership of companion animals has increased in Türkiye in recent years and that cats account for a large proportion of these animals, there is limited research on the vector-borne infectious agents carried by them. The present study aimed to provide a comprehensive molecular epidemiological data and molecular characterization of feline vector-borne haemopathogens (FVBHs), including piroplasms, anaplasmataceae, rickettsias, haemoplasmas, and Bartonella species in Türkiye. In total, 250 feline blood samples were collected from client-owned cats (n = 203) and shelter cats (n = 47) brought to the Small Animal Hospital of Selcuk University, Veterinary Faculty. RESULTS: Overall, 40 (16%) cats were found to be infected with at least one of the investigated haemopathogens and piroplasm, Mycoplasma spp. and Bartonella spp. prevalence was 1.6%, 11.2%, and 4.8%, respectively. No Anaplasma/Ehrlichia spp. and Rickettsia spp. DNA was detected in the investigated feline samples. Sequence analysis revealed that all four piroplasms belonged to Babesia ovis with a 97.93-99.82% nucleotide sequence identity to 18S rRNA gene sequences from Spain and Türkiye, while some sequenced hemoplasmas were Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm) and Mycoplasma wenyonii, and Bartonella spp. were Bartonella henselae and Bartonella koehlerae species. Co-infections with Mycoplasma spp. and Bartonella spp. were also detected in 4 cats (1.6%) in this study, where single infections were predominant. CONCLUSION: This study provides valuable information on zoonotically important feline vector-borne hemopathogens in Türkiye, some of which have received attention under the One Health perspective, and is the first molecular epidemiological study to demonstrate the presence of Babesia ovis, the causative agent of ovine babesiosis, and Mycoplasma wenyonii DNA, the causative agent of bovine haemotropic mycoplasmosis, in cats. Further studies on the roles of such pathogens detected in unspecific hosts and the host specificity of the vectors that transmit them will contribute to the elucidation of this situation.
Assuntos
Babesia , Doenças do Gato , Mycoplasma , Animais , Gatos , Doenças do Gato/parasitologia , Doenças do Gato/microbiologia , Doenças do Gato/epidemiologia , Mycoplasma/isolamento & purificação , Mycoplasma/genética , Babesia/isolamento & purificação , Babesia/genética , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/epidemiologia , Feminino , Masculino , Bartonella/isolamento & purificação , Bartonella/genética , Babesiose/epidemiologia , DNA Bacteriano , DNA de ProtozoárioRESUMO
In tropical regions, numerous tick-borne pathogens (TBPs) play a crucial role as causative agents of infectious diseases in humans and animals. Recently, the population of companion and pet dogs has significantly increased in Vietnam; however, information on the occurrence of TBPs is still limited. The objectives of this investigation were to determine the occurrence rate, risk factors, and phylogenetic characteristics of TBPs in dogs from northern Vietnam. Of 341 blood samples tested by PCR, the total infection of TBPs was 73.9% (252/341). Babesia vogeli (18SrRNA gene - 30.5%) was detected most frequently in studied dogs followed by Rickettsia spp. (OmpA gene - 27%), Anaplasma platys (groEL gene - 22%), Bartonella spp. (16SrRNA - 18.8%), Mycoplasma haemocanis (16SrRNA - 9.4%) and Hepatozoon canis (18SrRNA gene - 1.2%), respectively. All samples were negative for Ehrlichia canis and Anaplasma phagocytophylum. Co-infection was detected in 31.4% of the samples (107/341) of which, A. platys/Bartonella spp. (34/94,10%), Rickettsia spp./B. vogeli (19/94, 5.6%), and M. haemocanis/B. vogeli (19/94, 5.6%) were recorded as the three most frequent two species of co-infection types. Statistical analysis revealed a significant correlation between TBP infection and several host variables regarding age, breed, and living area in the current study. The recent findings reported herein, for the first time in Vietnam, are essential for local veterinarians when considering the appropriate approaches for diagnosing these diseases. Furthermore, this data can be used to establish control measures for future surveillance and prevention strategies against canine TBPs in Vietnam.
Assuntos
Anaplasma , Babesia , Doenças do Cão , Filogenia , Doenças Transmitidas por Carrapatos , Animais , Cães , Vietnã/epidemiologia , Doenças do Cão/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/microbiologia , Fatores de Risco , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/veterinária , Doenças Transmitidas por Carrapatos/microbiologia , Doenças Transmitidas por Carrapatos/parasitologia , Anaplasma/genética , Anaplasma/isolamento & purificação , Babesia/genética , Babesia/isolamento & purificação , Masculino , Feminino , Rickettsia/genética , Rickettsia/isolamento & purificação , Bartonella/genética , Bartonella/isolamento & purificação , Bartonella/classificação , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Mycoplasma/classificação , Coinfecção/veterinária , Coinfecção/epidemiologia , Coinfecção/parasitologia , Coinfecção/microbiologiaRESUMO
BACKGROUND: Hemotropic Mycoplasma species (hemoplasmas) cause hemolytic anemia in cats worldwide and are recognized as emerging zoonotic pathogens. There is no comprehensive study on the prevalence and species diversity of hemoplasmas in domestic cat populations in different regions in Iran. Thus, the aims of the present study were to provide data on the prevalence and molecular characterization of hemotropic Mycoplasma species in apparently healthy cats from six Iranian provinces with different climates. In addition, potential risk factors associated with hemoplasmosis in cats were assessed. RESULTS: Mycoplasma spp. DNA was detected in the blood of 56 / 361 cats (15.5%) using genus-specific PCR. Further examinations with species-specific PCR and Sanger sequencing showed that 38 cats (10.5%) tested positive for Candidatus Mycoplasma haemominutum (CMhm), 8 cats (2.2%) tested positive for Mycoplasma haemofelis (Mhf), and 2 cats (0.6%) tested positive for Candidatus Mycoplasma turicensis (CMt). Co-infection with CMhm, and Mhf was observed in 7 cats (1.9%). One cat (0.3%) showed mixed infection with CMhm, Mhf, and CMt. There were statistically significant relationships between Mycoplasma positivity and being female, living in shelter (cattery), and being over 3 years old (P < 0.05). No significant association was observed for the cat breed and sampling localities. CONCLUSIONS: Current study findings revealed that hemoplasma infections are common among Iran cat populations. Considering the impact of such emerging zoonotic pathogens on the One Health, routine screenings, increasing public awareness, effective control, and prophylactic strategies for minimizing infection in cats and subsequently in human are strongly recommended.
Assuntos
Doenças do Gato , DNA Bacteriano , Infecções por Mycoplasma , Mycoplasma , Filogenia , Animais , Gatos , Irã (Geográfico)/epidemiologia , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Doenças do Gato/microbiologia , Doenças do Gato/epidemiologia , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Mycoplasma/classificação , Prevalência , Feminino , Masculino , DNA Bacteriano/genética , Análise de Sequência de DNA , Reação em Cadeia da Polimerase , Fatores de Risco , Coinfecção/microbiologia , Coinfecção/veterinária , Coinfecção/epidemiologiaRESUMO
The present study aimed to investigate endothelial glycocalyx (eGCx) damage in cats with feline hemotropic mycoplasmosis caused by Mycoplasma haemofelis using selected biomarkers and to determine the diagnostic and prognostic significance of these biomarkers. The study included 25 cats with feline hemotropic mycoplasmosis and 10 healthy cats. Clinical examination, blood gas analysis, complete blood count, and biochemical analysis were performed. Hemotropic mycoplasmosis diagnosed by microscopic examination and molecularly confirmed by PCR targeting the Mycoplasma haemofelis 16s rRNA gene. To evaluate endothelial glycocalyx damage, syndecan-1, endothelin-1 (ET-1), asymmetric dimethylarginine (ADMA), and vascular endothelial growth factor-A (VEGF-A) concentrations were measured using cat-specific commercial ELISA kits. Of the cats with feline hemotropic mycoplasmosis, 14 (56%) survived and 11 (44%) died. While syndecan-1 and ET-1 concentrations were significantly higher in cats with hemotropic mycoplasmosis compared to the control group (p < 0.001), no statistically significant difference was found for ADMA and VEGF-A concentrations (p > 0.05). Endothelial glycocalyx biomarkers showed significant correlations with each other and with hematological parameters (p < 0.01). The results of the ROC analysis showed that ET-1 with area under the curve (AUC) of 0.821 (p < 0.01) and VEGF-A with AUC of 0.805 (p < 0.010) were found to be significant prognostic indicators. In conclusion, this study demonstrated that serum syndecan-1 and ET-1 can be used as diagnostic and serum ET-1 and VEGF-A as prognostic biomarkers in cats with hemotropic mycoplasmosis. Our results indicate the development of eGCx damage in feline hemotropic mycoplasmosis and suggest that glycocalyx disruption may contribute to the pathogenesis of the disease.
Assuntos
Biomarcadores , Doenças do Gato , Glicocálix , Mycoplasma , Fator A de Crescimento do Endotélio Vascular , Animais , Gatos , Glicocálix/metabolismo , Biomarcadores/sangue , Fator A de Crescimento do Endotélio Vascular/sangue , Doenças do Gato/microbiologia , Doenças do Gato/sangue , Doenças do Gato/diagnóstico , Mycoplasma/genética , Masculino , Feminino , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/sangue , Infecções por Mycoplasma/microbiologia , Endotelina-1/sangue , Sindecana-1/sangue , Arginina/análogos & derivados , Arginina/sangue , Arginina/metabolismoRESUMO
The Mycoplasma Immunoglobulin Binding/Protease (MIB-MIP) system is a candidate 'virulence factor present in multiple pathogenic species of the Mollicutes, including the fast-growing species Mycoplasma feriruminatoris. The MIB-MIP system cleaves the heavy chain of host immunoglobulins, hence affecting antigen-antibody interactions and potentially facilitating immune evasion. In this work, using -omics technologies and 5'RACE, we show that the four copies of the M. feriruminatoris MIB-MIP system have different expression levels and are transcribed as operons controlled by four different promoters. Individual MIB-MIP gene pairs of M. feriruminatoris and other Mollicutes were introduced in an engineered M. feriruminatoris strain devoid of MIB-MIP genes and were tested for their functionality using newly developed oriC-based plasmids. The two proteins are functionally expressed at the surface of M. feriruminatoris, which confirms the possibility to display large membrane-associated proteins in this bacterium. However, functional expression of heterologous MIB-MIP systems introduced in this engineered strain from phylogenetically distant porcine Mollicutes like Mesomycoplasma hyorhinis or Mesomycoplasma hyopneumoniae could not be achieved. Finally, since M. feriruminatoris is a candidate for biomedical applications such as drug delivery, we confirmed its safety in vivo in domestic goats, which are the closest livestock relatives to its native host the Alpine ibex.
Assuntos
Vacinas Bacterianas , Mycoplasma , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/genética , Mycoplasma/genética , Mycoplasma/imunologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Imunoglobulinas/genética , Imunoglobulinas/metabolismo , Imunoglobulinas/imunologia , Regulação Bacteriana da Expressão Gênica , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/imunologia , Infecções por Mycoplasma/prevenção & controle , CabrasRESUMO
Hemotropic mycoplasmas are bacteria that attaches to erythrocytes surface, which some species presents zoonotic concerns. In the suborder Pinnipedia, genera Otaria and Arctocephalus are prominent in Brazil. This study investigated the occurrence of hemoplasmas in Arctocephalus sp. and Otaria flavescens found dead along the coast of a Southern Brazilian State. DNA from 135 spleen samples were extracted and subjected to conventional PCR protocols, targeting the 16â¯S rRNA and 23â¯S rRNA gene. Three (2.22â¯%) Arctocephalus australis were positive in the 16â¯S rRNA gene, and no samples amplified in the 23â¯S rRNA gene. Samples from this study clustered with Zalophus californianus and Arctocephalus tropicalis mycoplasmas on a Bayesian phylogenetic analysis. Genetic diversity analysis suggested distinct genotypes, indicating A. australis as a new host for hemoplasma, and also a potential putative novel hemoplasma genotype. These findings raises future awareness for pinnipeds conservation, and adds Mycoplasma spp. to be taken into consideration when clinically evaluating rescued animals.
Assuntos
DNA Bacteriano , Otárias , Infecções por Mycoplasma , Mycoplasma , Filogenia , RNA Ribossômico 16S , Baço , Animais , Brasil/epidemiologia , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Mycoplasma/classificação , Otárias/microbiologia , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/epidemiologia , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Baço/microbiologia , RNA Ribossômico 23S/genética , Variação Genética , Genótipo , Teorema de Bayes , Autopsia/veterinária , Reação em Cadeia da PolimeraseRESUMO
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. METHODS: A 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. RESULTS: Flea 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. CONCLUSIONS: Therefore, 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.
Assuntos
Doenças do Gato , Ctenocephalides , Infecções por Mycoplasma , Mycoplasma , Animais , Mycoplasma/isolamento & purificação , Mycoplasma/genética , Mycoplasma/classificação , Ctenocephalides/microbiologia , Gatos , Doenças do Gato/parasitologia , Doenças do Gato/microbiologia , Doenças do Gato/diagnóstico , Doenças do Gato/transmissão , Doenças do Gato/epidemiologia , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/transmissão , Infecções por Mycoplasma/microbiologia , Infestações por Pulgas/veterinária , Infestações por Pulgas/parasitologia , Infestações por Pulgas/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , RNA Ribossômico 16S/genéticaRESUMO
PURPOSE: Trichomonas vaginalis is a causative agent of common non-viral sexually transmitted infections worldwide. However, the biological features, such as genotypes and endosymbionts, of T. vaginalis isolated in Japan remain unclear. The aim of this study was to characterize the actin-based genotypes and the endosymbionts of T. vaginalis isolated in Sapporo, Japan. METHODS: Three T. vaginalis clinical strains were isolated in Sapporo, Japan between 2019 and 2022. Actin-based genotyping was conducted by sequencing and phylogenetic analyses. The endosymbionts, such as Mycoplasma sp. and Trichomonasvirus, were detected using PCR and RT-PCR, respectively. Furthermore, the detected Mycoplasma spp. were identified using 16S rRNA gene sequencing. RESULTS: Of the three T. vaginalis strains, two belonged to genotype E, whereas one was genotype G as determined by actin-based genotyping. Two of the T. vaginalis strains harbored Mycoplasma spp. Using nearly full-length 16S rRNA gene sequencing, both were identified as Candidatus Mycoplasma girerdii. In contrast, the Trichomonasvirus was not found in the T. vaginalis strains. CONCLUSION: To our knowledge, this is the first report on the characterization of actin-based genotypes and the presence of endosymbiotic Ca. M. girerdii in T. vaginalis strains in Japan. Thus, this study will provide an important impetus for future research.
Assuntos
Actinas , Genótipo , Mycoplasma , Filogenia , RNA Ribossômico 16S , Simbiose , Trichomonas vaginalis , Trichomonas vaginalis/genética , Trichomonas vaginalis/isolamento & purificação , Japão , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Mycoplasma/classificação , Actinas/genética , Humanos , RNA Ribossômico 16S/genética , Feminino , Vaginite por Trichomonas/parasitologiaRESUMO
Introduction: Diagnosing Mycoplasma faucium poses challenges, and it's unclear if its rare isolation is due to infrequent occurrence or its fastidious nutritional requirements. Methods: This study analyzes the complete genome sequence of M. faucium, obtained directly from the pus of a sternum infection in a lung transplant patient using metagenomic sequencing. Results: Genome analysis revealed limited therapeutic options for the M. faucium infection, primarily susceptibility to tetracyclines. Three classes of mobile genetic elements were identified: two new insertion sequences, a new prophage (phiUMCG-1), and a species-specific variant of a mycoplasma integrative and conjugative element (MICE). Additionally, a Type I Restriction-Modification system was identified, featuring 5'-terminally truncated hsdS pseudogenes with overlapping repeats, indicating the potential for forming alternative hsdS variants through recombination. Conclusion: This study represents the first-ever acquisition of a complete circularized bacterial genome directly from a patient sample obtained from invasive infection of a primary sterile site using culture-independent, PCR-free clinical metagenomics.
Assuntos
Genoma Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica , Mycoplasma , Humanos , Metagenômica/métodos , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Mycoplasma/classificação , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/diagnóstico , Sequenciamento Completo do Genoma/métodos , Transplante de Pulmão , Prófagos/genética , Sequências Repetitivas Dispersas/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêuticoRESUMO
BACKGROUND: Having a simple and fast dividing organism capable of producing and exposing at its surface or secreting functional complex biomolecules with disulphide bridges is of great interest. The mycoplasma bacterial genus offers a set of relevant properties that make it an interesting chassis for such purposes, the main one being the absence of a cell wall. However, due to their slow growth, they have rarely been considered as a potential platform in this respect. This notion may be challenged with the recent discovery of Mycoplasma feriruminatoris, a species with a dividing time close to that of common microbial workhorses. So far, no tools for heterologous protein expression nor secretion have been described for it. RESULTS: The work presented here develops the fast-dividing M. feriruminatoris as a tool for secreting functional biomolecules of therapeutic interest that could be used for screening functional mutants as well as potentially for protein-protein interactions. Based on RNAseq, quantitative proteomics and promoter sequence comparison we have rationally designed optimal promoter sequences. Then, using in silico analysis, we have identified putative secretion signals that we validated using a luminescent reporter. The potential of the resulting secretion cassette has been shown with set of active clinically relevant proteins (interleukins and nanobodies). CONCLUSIONS: We have engineered Mycoplasma feriruminatoris for producing and secreting functional proteins of medical interest.
Assuntos
Proteínas de Bactérias , Mycoplasma , Mycoplasma/metabolismo , Mycoplasma/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Regiões Promotoras Genéticas , Proteômica , Anticorpos de Domínio Único/metabolismo , Anticorpos de Domínio Único/genéticaRESUMO
Mycoplasmas are atypical bacteria with small genomes that necessitate colonization of their respective animal or plant hosts as obligate parasites, whether as pathogens, or commensals. Some can grow axenically in specialized complex media yet show only host-cell-dependent growth in cell culture, where they can survive chronically and often through interactions involving surface colonization or internalization. To develop a mycoplasma-based system to identify genes mediating such interactions, we exploited genetically tractable strains of the goat pathogen Mycoplasma mycoides (Mmc) with synthetic designer genomes representing the complete natural organism (minus virulence factors; JCVI-syn1.0) or its reduced counterpart (JCVI-syn3B) containing only those genes supporting axenic growth. By measuring growth of surviving organisms, physical association with cultured human cells (HEK-293T, HeLa), and induction of phagocytosis by human myeloid cells (dHL-60), we determined that JCVI-syn1.0 contained a set of eight genes (MMSYN1-0179 to MMSYN1-0186, dispensable for axenic growth) conferring survival, attachment, and phagocytosis phenotypes. JCVI-syn3B lacked these phenotypes, but insertion of these genes restored cell attachment and phagocytosis, although not survival. These results indicate that JCVI-syn3B may be a powerful living platform to analyze the role of specific gene sets, from any organism, on the interaction with diverse mammalian cells in culture.
Assuntos
Mycoplasma mycoides , Mycoplasma , Animais , Humanos , Mycoplasma/genética , Mycoplasma mycoides/genética , Células HeLa , MamíferosRESUMO
BACKGROUND: Hemotropic 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. METHODS: One 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. RESULTS: The 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. CONCLUSIONS: Considering the infection rate of hemoplasmas shown in this study, further studies, such as on the pathogenicity and clinical significance of hemoplasmas are necessary.
Assuntos
Anemia , Doenças dos Bovinos , Infecções por Mycoplasma , Mycoplasma , Bovinos , Animais , Infecções por Mycoplasma/veterinária , Doenças dos Bovinos/epidemiologia , Mycoplasma/genética , Anemia/veterinária , RNA Ribossômico 16SRESUMO
Mycoplasma felis has been isolated from diseased cats and horses, but to date only a single fully assembled genome of this species, of an isolate from a horse, has been characterized. This study aimed to characterize and compare the completely assembled genomes of four clinical isolates of M. felis from three domestic cats, assembled with the aid of short- and long-read sequencing methods. The completed genomes encoded a median of 759 ORFs (range 743-777) and had a median average nucleotide identity of 98.2â% with the genome of the available equid origin reference strain. Comparative genomic analysis revealed the occurrence of multiple horizontal gene transfer events and significant genome reassortment. This had resulted in the acquisition or loss of numerous genes within the Australian felid isolate genomes, encoding putative proteins involved in DNA transfer, metabolism, DNA replication, host cell interaction and restriction modification systems. Additionally, a novel mycoplasma phage was detected in one Australian felid M. felis isolate by genomic analysis and visualized using cryo-transmission electron microscopy. This study has highlighted the complex genomic dynamics in different host environments. Furthermore, the sequences obtained in this work will enable the development of new diagnostic tools, and identification of future infection control and treatment options for the respiratory disease complex in cats.
Assuntos
Bacteriófagos , Felis , Mycoplasma , Gatos , Animais , Cavalos , Austrália , Genômica , Mycoplasma/genéticaRESUMO
Hemotropic mycoplasmas (hemoplasmas) are emerging zoonotic pathogens. Micromammals have received little attention as hosts for hemoplasmas despite their ubiquitous presence, high population abundances, and close association with humans. A PCR protocol targeting a fragment of the 16â¯S rRNA gene and direct sequencing in blood samples of 189 adult specimens and 35 fetuses belonging to three species of Eulipotyphla (shrews) and seven species of Rodentia, captured in three ecologically diverse habitats in North-Eastern Spain (Steppe, High Mountain, Mediterranean) yielded and occurrence of 26%, including 36% of 39 shrews and 23% of 150 rodents. Sequencing revealed the presence of 14 nucleotide sequence types (ntST) among the 56 readable sequences. In general, each ntST was associated with a given host species, although in some cases, the same ntST was sequenced in different species (chiefly rodents). Most ntST were closely related to rodent and/or bat hemoplasmas, but one was identical with Mycoplasma haemocanis/haemofelis, and others can be considered novel genotypes. High sequence diversity was detected in rodents, whereas in the white-toothed shrew (Crocidura russula), 9/11 sequences from two distant areas were identical. Phylogenetic and network analyses classified our sequences in different clades including hemoplasmas of rodents, carnivores, bats, and humans. Twelve of the fetuses (34.2%) of 9/12 litters (75.0%) of shrews and rodents were hemoplasma-positive, indicating frequent vertical transmission. Our study contributes to expanding our knowledge about the distribution, diversity, and transmission of hemoplasmas.
Assuntos
Carnívoros , Quirópteros , Infecções por Mycoplasma , Mycoplasma , Animais , Humanos , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/veterinária , Filogenia , Musaranhos/genética , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Mycoplasma/genética , Roedores , GenótipoRESUMO
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.
Assuntos
Canidae , Coinfecção , Infestações por Pulgas , Mycoplasma , Animais , Cães , Irã (Geográfico)/epidemiologia , Mycoplasma/genéticaRESUMO
A domestic short hair cat (Felis catus) suffering from a purulent wound infection resulting from a dog bite was sampled for bacterial culture and isolation as the wound had been unresponsive to prolonged antimicrobial treatment. A mycoplasma was isolated from the wound. Whole genome sequencing of the isolate was performed using short-read Illumina and long-read Oxford Nanopore chemistry, and the organism was identified as Mycoplasma edwardii. Comparison of the genome sequence of the isolate to a reference M. edwardii genome sequence (canid isolate) identified the loss of several key bacterial factors involved in genome editing, as well the insertion of several novel ORFs most closely related to those found in other canine mycoplasmas, specifically Mycoplasma canis, M. cynos, M. molare and M. maculosa. This is only the second known report of disease caused by M. edwardii in a non-canid species, and the first report of it infecting and causing clinical disease in a cat.
Assuntos
Infecções por Mycoplasma , Mycoplasma , Cães , Gatos , Animais , Infecções por Mycoplasma/veterinária , Infecções por Mycoplasma/microbiologia , Sistemas CRISPR-Cas , Transferência Genética Horizontal , Mycoplasma/genética , GenômicaRESUMO
Mycoplasma capricolum subspecies capripneumoniae (Mccp) is the causative agent of contagious caprine pleuropneumonia (CCPP), a devastating disease listed by the World Organisation for Animal Health (WOAH) as a notifiable disease and threatening goat production in Africa and Asia. Although a few commercial inactivated vaccines are available, they do not comply with WOAH standards and there are serious doubts regarding their efficacy. One of the limiting factors to comprehend the molecular pathogenesis of CCPP and develop improved vaccines has been the lack of tools for Mccp genome engineering. In this work, key synthetic biology techniques recently developed for closely related mycoplasmas were adapted to Mccp. CReasPy-Cloning was used to simultaneously clone and engineer the Mccp genome in yeast, prior to whole-genome transplantation into M. capricolum subsp. capricolum recipient cells. This approach was used to knock out an S41 serine protease gene recently identified as a potential virulence factor, leading to the generation of the first site-specific Mccp mutants. The Cre-lox recombination system was then applied to remove all DNA sequences added during genome engineering. Finally, the resulting unmarked S41 serine protease mutants were validated by whole-genome sequencing and their non-caseinolytic phenotype was confirmed by casein digestion assay on milk agar. The synthetic biology tools that have been successfully implemented in Mccp allow the addition and removal of genes and other genetic features for the construction of seamless targeted mutants at ease, which will pave the way for both the identification of key pathogenicity determinants of Mccp and the rational design of novel, improved vaccines for the control of CCPP.
Assuntos
Mycoplasma , Vacinas , Animais , Cabras , Mycoplasma/genética , Serina ProteasesRESUMO
Mycoplasma equirhinis is the predominant equine Mycoplasma sp. isolated from clinically normal horses and is suspected to be associated with inflammatory airway disease in which cough is the primary sign. Quantitative evaluation of bacterial counts is useful in assessing the association between the bacteria in samples and observed clinical signs, but this evaluation has been difficult with conventional culture methods of M. equirhinis given the need for pre-enrichment using liquid cultures. We established a quantitative real-time PCR (qPCR) assay for the quantification of M. equirhinis, targeting the hypothetical protein FJM08_00025. We confirmed its high species-specificity for M. equirhinis and a limit of detection of 2.9 copies/reaction. We quantified M. equirhinis in tracheal wash samples from 20 clinically normal horses and 22 coughing horses. The copy numbers detected by qPCR in 18 of the 22 samples from clinically affected horses were within the range detected in the 20 clinically normal horses (0-84 copies/reaction). The remaining 4 samples had considerably higher copy numbers (734-1,620,000 copies/reaction), suggesting the likely involvement of M. equirhinis infection. Quantitative evaluation of M. equirhinis over time using our qPCR assay may allow a more accurate assessment of M. equirhinis infection in coughing horses compared to culture methods.
Assuntos
Doenças dos Cavalos , Mycoplasma , Cavalos , Animais , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Mycoplasma/genética , Traqueia/microbiologia , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/microbiologiaRESUMO
Hemotrophic mycoplasmas (hemoplasmas) are epierythrocytic bacteria that infect wild and domestic animals, and can cause anemia in some of them. They are considered emerging and zoonotic pathogens, causing serious health problems in wildlife. Candidatus Mycoplasma haemolamae is the only species of hemoplasma that infects domestic South American camelids (alpacas and llamas), with limited studies in wild camelids. Therefore, the objective of this study was to determine the prevalence of Candidatus M. haemolamae in vicunas (Vicugna vicugna) from the Pampa Galeras National Reserve, located in the Ayacucho region of Peru, using molecular diagnosis. For this, blood samples from 79 vicunas were collected, which were molecularly analyzed by partially amplifying the 16S ribosomal RNA gene of Mycoplasma sp. Fourteen vicunas (17.7 %) were positive for the molecular diagnosis of Mycoplasma sp. All PCR-positive products were sequenced and showed more than 99 % identity with Candidatus M. haemolamae. Statistical analysis showed that tick-infested vicunas had 6.10 odds of presenting Candidatus M. haemolamae compared with tick-free vicunas. Sex and age were not associated with Candidatus M. haemolamae infections. This is the first report of hemoplasmas in vicunas, a wild South American camelid, demonstrating that the pathogen can have both a domestic and a wild life cycle. Future studies are necessary to know the current situation of this pathogen in domestic and wild camelids from other locations in Peru.