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1.
Aust Vet J ; 99(5): 163-171, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33751558

RESUMO

OBJECTIVE: Pleurisy in pigs has economic impacts in the production stage and at slaughter. This study sought to establish if some micro-organisms can be found in high numbers in lungs with pleurisy by assessing batches of pigs at an abattoir in Queensland Australia. DESIGN: Samples of lung (including trachea/bronchus and lymph nodes) from a maximum of 5 pleurisy affected pigs were collected from 46 batches of pigs representing 46 Queensland farms. PROCEDURE: Pleurisy-affected lung areas were cultured by traditional bacteriological methods and bacteria quantified by plate scores. Additionally, tracheal or bronchial swabs and apical lobe fluid were tested for Mycoplasma hyopneumoniae DNA and the superior tracheobronchial lymph nodes were tested for porcine circovirus type 2 DNA by polymerase chain reaction (PCR). All apparently significant bacteria were identified via PCR or sequencing. Typing was undertaken on some of the bacterial isolates. RESULTS: The most prevalent pathogens were M. hyopneumoniae, Streptococcus suis and Porcine Circovirus type 2, being found in 34, 38 and 31 batches, respectively. Other bacteria found were Actinobacillus species (29 batches), Pasteurella multocida (24 batches), Mycoplasma flocculare (9 batches), Actinobacillus pleuropneumoniae (7 batches), Mycoplasma hyorhinis (4 batches), Bisgaard Taxon 10 (1 batch), Glaesserella parasuis (1 batch), Streptococcus minor (1 batch) and Streptococcus porcinus (1 batch). Most batches had more than one bacterial species. CONCLUSION: The high percentage of batches infected with S. suis (83%), M. hyopneumoniae (74%) and PCV2 (70%) and clustering by a batch of these pathogens, as well as the presence of many secondary pathogens, suggests synergy between these organisms may have resulted in pleurisy.


Assuntos
Pleurisia , Doenças dos Suínos , Matadouros , Animais , Austrália/epidemiologia , Pulmão , Mycoplasma , Pleurisia/epidemiologia , Pleurisia/veterinária , Queensland/epidemiologia , Streptococcus , Suínos , Doenças dos Suínos/epidemiologia
2.
J Zoo Wildl Med ; 51(4): 879-888, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33480568

RESUMO

Ophthalmic studies of the Texas tortoise (Gopherus berlandieri) established normal ophthalmic parameters for select diagnostic tests in captive tortoises and assessment of differences among individuals of differing size and health status. Sixty-one tortoises of varying weight, shell size, Mycoplasma seroprevalence, and herpesvirus exposure were included. Complete ophthalmic examinations, including neuro-ophthalmic reflexes, phenol red thread test, rebound tonometry, fluorescein staining, palpebral fissure length measurement, slit lamp biomicroscopy, indirect fundoscopy, and ocular ultrasound measurements of axial globe length, anterior chamber depth, lens thickness, and vitreous length, were recorded. All tortoises had negative dazzle and pupillary light reflexes, inconsistent menace responses, and positive palpebral reflexes. Mean ± SD tear production and intraocular pressure (IOP) were 14.2 ± 5.6 mm/15 sec and 13.8 ± 2.4 mm Hg in healthy tortoises, respectively. Mycoplasma-seropositive tortoises (with or without herpesvirus exposure) had significantly increased tear production (20.2 ± 8.1 and 19.9 ± 8.9 mm/15 sec, respectively) compared with healthy seronegative tortoises (14.2 ± 5.6 mm/15 sec; P = 0.02). As body size decreased, so too did palpebral fissure length and ocular ultrasound measurements, while IOP increased. Overall, palpebral fissure length appeared relatively small, and tear production relatively increased compared with other chelonian species, likely on the basis of the relatively arid native habitat. Further work is recommended to establish baseline values in related species, as well as comparison in aquatic versus terrestrial chelonians. The authors further suggest that the finding of relatively increased tear production in tortoises may indicate the need to rule out mycoplasmosis as a cause of upper respiratory tract disease.


Assuntos
Olho/anatomia & histologia , Tartarugas/anatomia & histologia , Animais , Oftalmopatias/patologia , Feminino , Masculino , Mycoplasma/isolamento & purificação , Lágrimas , Tonometria Ocular
3.
Korean J Parasitol ; 58(5): 565-569, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33202509

RESUMO

This report describes the first clinical case of a transfusion-associated Mycoplasma haemocanis infection in a dog in Korea. A 6-year-old male Maltese underwent a red blood cell transfusion for idiopathic immune-mediated hemolytic anemia. Eighteen days after the blood transfusion, the recipient's packed cell volume decreased and basophilic organisms were found on erythrocytes. A polymerase chain reaction and sequential analysis showed that both the donor dog and recipient dog had M. haemocanis. Six weeks after doxycycline administration, no organisms were detected and the recipient's anemia had improved.


Assuntos
Anemia Hemolítica Autoimune/terapia , Anemia Hemolítica Autoimune/veterinária , Transfusão de Sangue/veterinária , Doenças do Cão/terapia , Doenças do Cão/transmissão , Doxiciclina/administração & dosagem , Infecções por Mycoplasma/transmissão , Infecções por Mycoplasma/veterinária , Mycoplasma , Reação Transfusional/microbiologia , Reação Transfusional/veterinária , Animais , Doenças do Cão/etiologia , Doenças do Cão/microbiologia , Cães , Masculino , Infecções por Mycoplasma/tratamento farmacológico , Infecções por Mycoplasma/microbiologia , República da Coreia , Resultado do Tratamento
4.
Rev Bras Parasitol Vet ; 29(3): e007320, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32935770

RESUMO

Although anemia has been historically linked to Haemonchus contortus infection, other infectious agents, such as hemotropic mycoplasmas and tick-borne disease pathogens, may also lead to anemic crisis in sheep. This study has aimed to investigate infections related to anemia in a sheep herd from Bandeirantes City, Paraná State, southern Brazil. Seven out of forty-two (16.6%; 95% CI: 8.32-30.6%) sheep were positive for hemoplasmas by a PCR targeting the 16S rRNA gene and all tested negative for A. marginale/A. ovis and Babesia/Theileria spp. by PCR based on msp4 and 18S rRNA genes, respectively. Two (4.7%; 95% CI: 1.32-15.79%) animals were infested with Rhipicephalus microplus ticks. Fecal egg counting was performed in 38 sheep and 24 (63.15%; 95% CI: 47.2-76.6%) presented > 500 eggs per gram. Phylogenetic analysis of partial sequences of the detected hemotropic Mycoplasma sp. 16S and 23S rRNA genes confirmed that the animals were infected with Mycoplasma ovis. Polymorphism analysis of partial 16S rRNA sequences showed three different genotypes of M. ovis infecting sheep assessed in the present study. Mycoplasma ovis and gastrointestinal nematodes occurs in sheep from the northern region of Paraná State.


Assuntos
Anemia/veterinária , Nematoides , Parasitos , Doenças Parasitárias em Animais , Doenças dos Ovinos , Anemia/complicações , Anemia/parasitologia , Animais , Brasil , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Nematoides/isolamento & purificação , Parasitos/classificação , Parasitos/isolamento & purificação , Doenças Parasitárias em Animais/complicações , Doenças Parasitárias em Animais/microbiologia , Doenças Parasitárias em Animais/parasitologia , Filogenia , RNA Ribossômico 16S/genética , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/parasitologia , Inquéritos e Questionários
5.
PLoS One ; 15(8): e0238202, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32846428

RESUMO

The immune system of ectotherms, particularly non-avian reptiles, remains poorly characterized regarding the genes involved in immune function, and their function in wild populations. We used RNA-Seq to explore the systemic response of Mojave desert tortoise (Gopherus agassizii) gene expression to three levels of Mycoplasma infection to better understand the host response to this bacterial pathogen. We found over an order of magnitude more genes differentially expressed between male and female tortoises (1,037 genes) than differentially expressed among immune groups (40 genes). There were 8 genes differentially expressed among both variables that can be considered sex-biased immune genes in this tortoise. Among experimental immune groups we find enriched GO biological processes for cysteine catabolism, regulation of type 1 interferon production, and regulation of cytokine production involved in immune response. Sex-biased transcription involves iron ion transport, iron ion homeostasis, and regulation of interferon-beta production to be enriched. More detailed work is needed to assess the seasonal response of the candidate genes found here. How seasonal fluctuation of testosterone and corticosterone modulate the immunosuppression of males and their susceptibility to Mycoplasma infection also warrants further investigation, as well as the importance of iron in the immune function and sex-biased differences of this species. Finally, future transcriptional studies should avoid drawing blood from tortoises via subcarapacial venipuncture as the variable aspiration of lymphatic fluid will confound the differential expression of genes.


Assuntos
Infecções por Mycoplasma/imunologia , Infecções por Mycoplasma/veterinária , Mycoplasma/imunologia , Tartarugas/genética , Tartarugas/imunologia , Animais , Anticorpos Antibacterianos/sangue , California , Citocinas/genética , Citocinas/imunologia , Clima Desértico , Feminino , Regulação da Expressão Gênica/genética , Regulação da Expressão Gênica/imunologia , Interferon Tipo I/genética , Interferon Tipo I/imunologia , Transporte de Íons/genética , Ferro/metabolismo , Masculino , Infecções por Mycoplasma/microbiologia , Nevada , Fatores Sexuais
6.
J Anim Sci ; 98(8)2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32717077

RESUMO

Developments of pulmonary diseases, often accompanied by infections of bacteria, severely affect the meat production and welfare of pigs. This study investigated 307 pigs at age of 240 d from an eight-breed cross reared under standardized housing conditions for associations among the extent of lung lesions, bacteria load inferred from 16S rRNA sequencing of bronchoalveolar lavage fluid, as well as 57 immune cells and 25 hematological traits. We showed that the pigs under study suffered substantial and varied lung lesions, and the Mycoplasma is the most associated bacteria genera. At a false discovery rate of 0.05 (FDR < 0.05), the severity of lung lesions were significantly associated with greater CD8+ to CD3+ cell ratio, neutrophil-to-lymphocyte ratio (NLR), and standard deviation of red blood cell volume distribution width (RDW-SD), and lower CD4-CD8-/CD3+, CD3+CD4-CD8-/PBMCs (peripheral blood mononuclear cells) and CD14-CD16-/PBMCs cell ratios, mean corpuscular hemoglobin concentration, lymphocyte count, and lymphocyte count percentage, reflecting an status of inflammation, immune suppression, and hypoxia of the pigs accompanying the progression of the lung lesions. The Mycoplasma abundance showed positive correlations with neutrophil count, neutrophil count percentage, NLR, monocyte count, coefficient of variation in red blood cell volume distribution width , and RDW-SD, and negative correlations with mean corpuscular hemoglobin concentration, lymphocyte count, and lymphocyte count percentage; these correlations are largely consistent with those of lung lesions, supporting the comorbidity of lung lesions and Mycoplasma infection. We also observed nonlinear associations that sharp increases in neutrophil count and neutrophil count percentage occurred only when Mycoplasma abundance raised above the population-average level. The results provide helpful insights into the changes of host immune status in response to Mycoplasma relevant lung diseases in pigs.


Assuntos
Carga Bacteriana , Infecções por Mycoplasma/veterinária , Doenças dos Suínos/microbiologia , Animais , Índices de Eritrócitos/veterinária , Predisposição Genética para Doença , Inflamação/patologia , Inflamação/veterinária , Contagem de Leucócitos/veterinária , Leucócitos Mononucleares , Pulmão/patologia , Contagem de Linfócitos/veterinária , Linfócitos , Mycoplasma/imunologia , Infecções por Mycoplasma/genética , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/patologia , Neutrófilos , RNA Ribossômico 16S , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/patologia
7.
Sci Rep ; 10(1): 7641, 2020 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-32376831

RESUMO

Lcn2 gene expression increases in response to cell stress signals, particularly in cells involved in the innate immune response. Human Lcn2 (NGAL) is increased in the blood and tissues in response to many stressors including microbial infection and in response to LPS in myeloid and epithelial cells. Here we extend the microbial activators of Lcn2 to mycoplasma and describe studies in which the mechanism of Lcn2 gene regulation by MALP-2 and mycoplasma infection was investigated in mouse mammary epithelial cells. As for the LPS response of myeloid cells, Lcn2 expression in epithelial cells is preceded by increased TNFα, IL-6 and IκBζ expression and selective reduction of IκBζ reduces Lcn2 promoter activity. Lcn2 promoter activation remains elevated well beyond the period of exposure to MALP-2 and is persistently elevated in mycoplasma infected cells. Activation of either the human or the mouse Lcn2 promoter requires both NFκB and C/EBP for activation. Thus, Lcn2 is strongly and enduringly activated by mycoplasma components that stimulate the innate immune response with the same basic regulatory mechanism for the human and mouse genes.


Assuntos
Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Células Epiteliais/metabolismo , Lipocalina-2/genética , Infecções por Mycoplasma/genética , Infecções por Mycoplasma/metabolismo , Infecções por Mycoplasma/microbiologia , Mycoplasma/fisiologia , NF-kappa B/metabolismo , Animais , Sítios de Ligação , Regulação da Expressão Gênica , Interações Hospedeiro-Patógeno , Humanos , Lipopeptídeos/metabolismo , Camundongos , Ligação Proteica , Transdução de Sinais
8.
BMC Vet Res ; 16(1): 107, 2020 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-32252763

RESUMO

BACKGROUND: Mycoplasmas primarily cause respiratory or urogenital tract infections impacting avian, bovine, canine, caprine, murine, and reptilian hosts. In animal husbandry, mycoplasmas cause reduced feed-conversion, decreased egg production, arthritis, hypogalactia or agalactia, increased condemnations, culling, and mortality in some cases. Antibiotics reduce transmission and mitigate clinical signs; however, concerning levels of antibiotic resistance in Mycoplasma gallisepticum and M. capricolum isolates exist. To address these issues, we evaluated the minimum inhibitory concentrations (MICs) of halogenated phenazine and quinoline compounds, an N-arylated NH125 analogue, and triclosan against six representative veterinary mycoplasmas via microbroth or agar dilution methods. Thereafter, we evaluated the minimum bactericidal concentration (MBC) of efficacious drugs. RESULTS: We identified several compounds with MICs ≤25 µM against M. pulmonis (n = 5), M. capricolum (n = 4), M. gallisepticum (n = 3), M. alligatoris (n = 3), M. agassizii (n = 2), and M. canis (n = 1). An N-arylated NH125 analogue, compound 21, served as the most efficacious, having a MIC ≤25 µM against all mycoplasmas tested, followed by two quinolines, nitroxoline (compound 12) and compound 20, which were effective against four and three mycoplasma type strains, respectively. Nitroxoline exhibited bactericidal activity among all susceptible mycoplasmas, and compound 21 exhibited bactericidal activity when the MBC was able to be determined. CONCLUSIONS: These findings highlight a number of promising agents from novel drug classes with potential applications to treat veterinary mycoplasma infections and present the opportunity to evaluate preliminary pharmacokinetic indices using M. pulmonis in rodents as an animal model of human infection.


Assuntos
Antibacterianos/farmacologia , Imidazóis/farmacologia , Mycoplasma/efeitos dos fármacos , Fenazinas/farmacologia , Quinolinas/farmacologia , Testes de Sensibilidade Microbiana
9.
Acta Trop ; 207: 105500, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32330451

RESUMO

In the scientific literature, a small amount of information is found concerning mycoplasmosis in camel species. A variety of pathogens could be causative agents for pneumonia, but walking pneumonia is mostly caused by Mycoplasma with slow development and mild symptoms. The aim of this study was to identify mycoplasmas from camels (Camelus dromedarius) and extending the arsenal of factors implicated in pathogenicity of M. arginini to shed light on the current knowledge gap. 460 lung samples (pneumonic; n=210 and apparently healthy; n=250) were randomly collected from the one-humped camels (C. domedarius) that have been imported from Sudan and slaughtered at Cairo Slaughterhouse. 48 out of 210 isolates (22.9%) recovered from the pneumonic lungs were recorded as M. arginini. Positive PCR results were obtained for all 48 isolates. On the other hand, infection with the organism was not detected in the apparently healthy lungs. Hemolysis and hydrogen sulphide (H2S) production, a compound that has previously not been identified as a virulence factor in M. arginini, was evident in 100% of the isolates. The 48 M. arginini isolates were weak in their ability to form biofilm on polystyrene surfaces. All isolates were 100% susceptible to florfenicol and streptomycin and 100% resistant to ciprofloxacin. Resistance to lincomycin, spiromycin, tylosin, doxacyclin and erythromycin was observed at different frequencies. 13 different combinations of antibiotics representing one to four classes were evident with the Macrolide erythromycin being the most represented. It also should be noted that the ciprofloxacin, doxacyclin, lincomycin, erythromycin combination was the most noted in 21/48 isolates. Surprisingly, none of the virulence genes (vsp, uvrC and gapA) and quinolone resistance genes (parC and gyrA) were detected by PCR.


Assuntos
Camelus/microbiologia , Mycoplasma/isolamento & purificação , Animais , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Mycoplasma/efeitos dos fármacos , Mycoplasma/genética , Mycoplasma/patogenicidade , Filogenia , Virulência/genética
10.
PLoS One ; 15(4): e0231239, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32294121

RESUMO

BACKGROUND: Chorioamnionitis has been linked to spontaneous preterm labor and complications such as neonatal sepsis. We hypothesized that microbial cell-free (cf) DNA would be detectable in maternal plasma in patients with chorioamnionitis and could be the basis for a non-invasive method to detect fetal exposure to microorganisms. OBJECTIVE: The purpose of this study was to determine whether next generation sequencing could detect microbial cfDNA in maternal plasma in patients with chorioamnionitis. STUDY DESIGN: Maternal plasma (n = 94) and umbilical cord plasma (n = 120) were collected during delivery at gestational age 28-41 weeks. cfDNA was extracted and sequenced. Umbilical cord plasma samples with evidence of contamination were excluded. The prevalence of microorganisms previously implicated in choriomanionitis, neonatal sepsis and intra-amniotic infections, as described in the literature, were examined to determine if there was enrichment of these microorganisms in this cohort. Specific microbial cfDNA associated with chorioamnionitis was first detected in umbilical cord plasma and confirmed in the matched maternal plasma samples (n = 77 matched pairs) among 14 cases of histologically confirmed chorioamnionitis and one case of clinical chorioamnionitis; 63 paired samples were used as controls. A correlation of rank of a given microorganism across maternal plasma and matched umbilical cord plasma was used to assess whether signals found in umbilical cord plasma were also present in maternal plasma. RESULTS: Microbial DNA sequences associated with clinical and/or histological chorioamnionitis were enriched in maternal plasma in cases with suspected chorioamnionitis when compared to controls (12/14 microorganisms, p = 0.02). Analysis of the microbial cfDNA in umbilical cord plasma among the 1,251 microorganisms detectable with this assay identified Streptococcus mitis, Ureaplasma spp., and Mycoplasma spp. in cases of suspected chorioamnionitis. This assay also detected cfDNA from Lactobacillus spp. in controls. Comparison between maternal plasma and umbilical cord plasma confirmed these signatures were also present in maternal plasma. Unbiased analysis of microorganisms with significantly correlated signal between matched maternal plasma and umbilical cord plasma identified the above listed 3 microorganisms, all of which have previously been implicated in patients with chorioamnionitis (Mycoplasma hominis p = 0.0001; Ureaplasma parvum p = 0.002; Streptococcus mitis p = 0.007). These data show that the pathogen signal relevant for chorioamnionitis can be identified in both maternal and umbilical cord plasma. CONCLUSION: This is the first report showing the detection of relevant microbial cell-free cfDNA in maternal plasma and umbilical cord plasma in patients with clinical and/or histological chorioamnionitis. These results may lead to the development of a specific assay to detect perinatal infections for targeted therapy to reduce early neonatal sepsis complications.


Assuntos
Ácidos Nucleicos Livres/sangue , Corioamnionite/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Análise de Sequência de DNA/métodos , Cordão Umbilical/microbiologia , Adulto , Corioamnionite/microbiologia , Estudos de Coortes , Feminino , Sangue Fetal/química , Sangue Fetal/metabolismo , Sangue Fetal/microbiologia , Idade Gestacional , Humanos , Recém-Nascido , Mycoplasma/genética , Mycoplasma/patogenicidade , Sepse Neonatal/sangue , Sepse Neonatal/diagnóstico , Sepse Neonatal/microbiologia , Gravidez , Streptococcus mitis/genética , Streptococcus mitis/patogenicidade , Cordão Umbilical/patologia , Ureaplasma/genética , Ureaplasma/patogenicidade , Adulto Jovem
11.
Appl Environ Microbiol ; 86(12)2020 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-32276983

RESUMO

Mycoplasma haemocanis is prevalent in the endangered Darwin's fox (Lycalopex fulvipes) in its main stronghold, Chiloé Island (Chile). The origin of the infection, its dynamics, its presence in other fox populations and the potential consequences for fox health remain unexplored. For 8 years, hemoplasmal DNA was screened and characterized in blood from 82 foxes in Chiloé and two other fox populations and in 250 free-ranging dogs from Chiloé. The prevalence of M. haemocanis in foxes was constant during the study years, and coinfection with "Candidatus Mycoplasma haematoparvum" was confirmed in 30% of the foxes. Both hemoplasma species were detected in the two mainland fox populations and in Chiloé dogs. M. haemocanis was significantly more prevalent and more genetically diverse in foxes than in dogs. Two of the seven M. haemocanis haplotypes identified were shared between these species. Network analyses did not show genetic structure by species (foxes versus dogs), geographic (island versus mainland populations), or temporal (years of study) factors. The probability of infection with M. haemocanis increased with fox age but was not associated with sex, season, or degree of anthropization of individual fox habitats. Some foxes recaptured years apart were infected with the same haplotype in both events, and no hematological alterations were associated with hemoplasma infection, suggesting tolerance to the infection. Altogether, our results indicate that M. haemocanis is enzootic in the Darwin's fox and that intraspecific transmission is predominant. Nevertheless, such a prevalent pathogen in a threatened species represents a concern that must be considered in conservation actions.IMPORTANCE Mycoplasma haemocanis is enzootic in Darwin's foxes. There is a higher M. haemocanis genetic diversity and prevalence in foxes than in sympatric dogs, although haplotypes are shared between the two carnivore species. There is an apparent tolerance of Darwin's foxes to Mycoplasma haemocanis.


Assuntos
Infecções Assintomáticas/epidemiologia , Coinfecção/veterinária , Doenças do Cão/epidemiologia , Raposas , Infecções por Mycoplasma/veterinária , Mycoplasma/isolamento & purificação , Animais , Chile/epidemiologia , Coinfecção/epidemiologia , Coinfecção/microbiologia , Doenças do Cão/microbiologia , Cães , Espécies em Perigo de Extinção , Feminino , Masculino , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Prevalência
12.
Res Vet Sci ; 130: 139-143, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32182439

RESUMO

MALDI-TOF MS is a fast and accurate tool to identify Mycoplasma species in liquid media. However, when trying to identify presumptive Mycoplasma bovis (M. bovis) colonies from solid medium (the "direct transfer method") a surprisingly high occurrence of M. arginini and M. alkalescens identification was observed. It was hypothesized that agar medium components are associated with false positive identification with Mycoplasma spp., as M. bovis colonies are very small and grow into the agar. The objective of this study was to determine whether complete modified pleuropneumonia-like organism (PPLO) agar (supplemented with horse serum, sodium pyruvate, technical yeast extract, ampicillin sodium salt and colistin) and the separate components, result in false identification as Mycoplasma spp. by MALDI-TOF MS. A total of 100 samples were examined, of which 33% of the modified PPLO agar spots were identified as M. alkalescens (16%) and M. arginini (17%)), albeit with relatively low score values (< 1.85). No false identification of M. bovis was obtained. Several medium components (unsupplemented PPLO agar, horse serum and colistin) resulted in spectra with peaks showing close matches with peaks present in the M. alkalescens and M. arginini database spectra. This study shows that the direct transfer method should be interpreted with caution, and one should strive to pick as little as possible agar when sampling Mycoplasma-like colonies from solid medium containing PPLO agar, horse serum and/or colistin.


Assuntos
Ágar/química , Doenças dos Bovinos/diagnóstico , Meios de Cultura/química , Infecções por Mycoplasma/veterinária , Mycoplasma/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Testes Diagnósticos de Rotina/veterinária , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/microbiologia , Manejo de Espécimes/veterinária
13.
Vet Res ; 51(1): 46, 2020 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-32209128

RESUMO

Infection with Bordetella bronchiseptica (Bb), a pathogen involved in canine infectious respiratory disease complex, can be confirmed using culture or qPCR. Studies about the canine lung microbiota (LM) are recent, sparse, and only one paper has been published in canine lung infection. In this study, we aimed to compare the LM between Bb infected and healthy dogs, and to correlate sequencing with culture and qPCR results. Twenty Bb infected dogs diagnosed either by qPCR and/or culture and 4 healthy dogs were included. qPCR for Mycoplasma cynos (Mc) were also available in 18 diseased and all healthy dogs. Sequencing results, obtained from bronchoalveolar lavage fluid after DNA extraction, PCR targeting the V1-V3 region of the 16S rDNA and sequencing, showed the presence of Bb in all diseased dogs, about half being co-infected with Mc. In diseased compared with healthy dogs, the ß-diversity changed (P = 0.0024); bacterial richness and α-diversity were lower (P = 0.012 and 0.0061), and bacterial load higher (P = 0.004). Bb qPCR classes and culture results correlated with the abundance of Bb (r = 0.71, P < 0.001 and r = 0.70, P = 0.0022). Mc qPCR classes also correlated with the abundance of Mc (r = 0.73, P < 0.001). Bb infection induced lung dysbiosis, characterized by high bacterial load, low richness and diversity and increased abundance of Bb, compared with healthy dogs. Sequencing results highly correlate with qPCR and culture results showing that sequencing can be reliable to identify microorganisms involved in lung infectious diseases.


Assuntos
Carga Bacteriana , Infecções por Bordetella/veterinária , Bordetella bronchiseptica/isolamento & purificação , Doenças do Cão/microbiologia , Pulmão/microbiologia , Infecções Respiratórias/veterinária , Animais , Infecções por Bordetella/microbiologia , Coinfecção/microbiologia , Coinfecção/veterinária , Cães , Microbiota , Mycoplasma/isolamento & purificação , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Infecções Respiratórias/microbiologia
14.
Pesqui. vet. bras ; 40(3): 220-225, Mar. 2020. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1135611

RESUMO

Brazil is one of the countries with the most abundant avifauna in the world. The confinement of birds associated with close contact with other animals and humans favor the spread of agents of respiratory diseases. Among them, mycoplasmas can cause asymptomatic or apparent disease that manifests in birds by coughing, sneezing, rales, conjunctivitis, ocular and nasal discharge. Several described mycoplasmas cause disease in birds, especially Mycoplasma gallisepticum(MG) andMycoplasma synoviae(MS). The diagnosis ofMycoplasmaspp. can be done by clinical observation and laboratory analysis. Molecular diagnosis by PCR was boosted by its speed, sensitivity, and low cost of agent isolation techniques that take up to 21 days to complete. This study aimed to verify the occurrence ofMycoplasmaspp. in birds of the Rio de Janeiro Zoo (Rio Zoo), by isolation and PCR. Of the total 635 birds from the Rio Zoo, 81 were studied for detection ofMycoplasmaspp., when taken for routine health assessment exams. These birds belonged to the following orders: Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) and Cariamiformes (1), all individuals already identified by microchip or leg-ring. There was no isolation of mycoplasmas in any of the samples tested, whereas, in the PCR, 62.96% (51/81) were positive, with 1.96% (1/51) identified as MG and 19.61% (10/51) as MS, representing 1.23% (1/81) and 12.34% (10/81) of the total population studied. PCR was shown to be a more effective technique than isolation in the detection ofMycoplasmaspp. in birds. It was possible to detect mycoplasmas in birds from Riozoo with no clinical respiratory signs, with higher MS prevalence than MG. The positivities forMycoplasmaspp., MS, and MG were different among the orders studied, being the highest occurrence in birds of prey, followed by Galliformes and Piciformes. The presence of MG and MS in birds of Rio de Janeiro Zoo confirms the circulation of these agents and the need for further studies on the dissemination of mycoplasmas in zoos for the epidemiological analysis of these bacteria in these places.(AU)


O Brasil é um dos países com maior avifauna do mundo. O confinamento de aves associado ao contato próximo a outros animais e seres humanos favorece a disseminação de agentes etiológicos causadores de doenças respiratórias. Dentre eles, os micoplasmas podem causar doença assintomática ou aparente que se manifesta em aves por espirros, estertores, conjuntivite, corrimentos oculares e nasais. São diversos os micoplasmas descritos causadores de doença em aves, com destaque para Mycoplasma gallisepticum (MG) e Mycoplasma synoviae (MS). O diagnóstico de Mycoplasma spp. pode ser feito pela observação clínica e análises laboratoriais. O diagnóstico molecular pela Reação em Cadeia da Polimerase (PCR) ganhou impulso por sua rapidez, sensibilidade e baixo custo em relação às técnicas de isolamento do agente que levam até 21 dias para conclusão do gênero Mycoplasma. Objetivou-se verificar a ocorrência da infecção por Mycoplasma spp. em aves no Zoológico do Rio de Janeiro (Rio Zoo), por isolamento e PCR. Do plantel de 635 aves do Rio Zoo, foram estudadas 81 para detecção de Mycoplasma spp., quando contidas para exames rotineiros de avaliação da condição de saúde. Essas aves eram pertencentes às ordens Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) e Cariamiformes (1), todas já identificadas por microchip ou por anilha. Não houve isolamento de micoplasmas em nenhuma das amostras testadas, enquanto na PCR, 62,96% (51/81) foram positivas, sendo 1,96% (1/51) identificadas como MG e 19,61% (10/51) como MS, representando 1,23% (1/81) e 12,34% (10/81) da população total estudada. A PCR demonstrou ser uma técnica mais efetiva que o isolamento na detecção de Mycoplasma spp. em aves. Foi possível detectar micoplasmas nas aves do Riozoo sem sinal clínico respiratório, tendo MS maior prevalência do que MG. As positividades para Mycoplasma spp., MG e MS foram diferentes entre as ordens de aves estudadas, sendo a maior ocorrência nas aves de rapina, seguida dos Galliformes e dos Piciformes. A presença de MG e MS nas aves do Rio de Janeiro Zoo confirma a circulação destes agentes e a necessidade de mais estudos sobre a disseminação de micoplasmas em zoológicos para análise epidemiológica dessas bactérias nesse local.(AU)


Assuntos
Animais , Psittaciformes/microbiologia , Aves Predatórias/microbiologia , Mycoplasma gallisepticum/isolamento & purificação , Mycoplasma synoviae/isolamento & purificação , Galliformes/microbiologia , Animais de Zoológico/microbiologia , Mycoplasma/isolamento & purificação , Infecções por Mycoplasma/epidemiologia , Aves/microbiologia , Reação em Cadeia da Polimerase/veterinária
15.
Int J Mol Sci ; 21(4)2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-32075244

RESUMO

Studies of the human microbiome have elucidated an array of complex interactions between prokaryotes and their hosts. However, precise bacterial pathogen-cancer relationships remain largely elusive, although several bacteria, particularly those establishing persistent intra-cellular infections, like mycoplasmas, can alter host cell cycles, affect apoptotic pathways, and stimulate the production of inflammatory substances linked to DNA damage, thus potentially promoting abnormal cell growth and transformation. Consistent with this idea, in vivo experiments in several chemically induced or genetically deficient mouse models showed that germ-free conditions reduce colonic tumor formation. We demonstrate that mycoplasma DnaK, a chaperone protein belonging to the Heath shock protein (Hsp)-70 family, binds Poly-(ADP-ribose) Polymerase (PARP)-1, a protein that plays a critical role in the pathways involved in recognition of DNA damage and repair, and reduces its catalytic activity. It also binds USP10, a key p53 regulator, reducing p53 stability and anti-cancer functions. Finally, we showed that bystander, uninfected cells take up exogenous DnaK-suggesting a possible paracrine function in promoting cellular transformation, over and above direct mycoplasma infection. We propose that mycoplasmas, and perhaps certain other bacteria with closely related DnaK, may have oncogenic activity, mediated through the inhibition of DNA repair and p53 functions, and may be involved in the initiation of some cancers but not necessarily involved nor necessarily even be present in later stages.


Assuntos
Inflamação/genética , Chaperonas Moleculares/genética , Infecções por Mycoplasma/genética , Mycoplasma/genética , Neoplasias/genética , Apoptose/genética , Transformação Celular Neoplásica/genética , Dano ao DNA/genética , Reparo do DNA/genética , Humanos , Inflamação/microbiologia , Inflamação/patologia , Mycoplasma/patogenicidade , Infecções por Mycoplasma/microbiologia , Neoplasias/patologia , Poli(ADP-Ribose) Polimerase-1/genética , Proteína Supressora de Tumor p53/genética , Ubiquitina Tiolesterase/genética
17.
Int J Syst Evol Microbiol ; 70(4): 2369-2381, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32068526

RESUMO

In 1983, Mycoplasma sp. strain 1220 was isolated in Hungary from the phallus lymph of a gander with phallus inflammation. Between 1983 and 2017, Mycoplasma sp. 1220 was also identified and isolated from the respiratory tract, liver, ovary, testis, peritoneum and cloaca of diseased geese in several countries. Seventeen studied strains produced acid from glucose and fructose but did not hydrolyse arginine or urea, and all grew under aerobic, microaerophilic and anaerobic conditions at 35 to 37 ˚C in either SP4 or pleuropneumonia-like organism medium supplemented with glucose and serum. Colonies on agar showed a typical fried-egg appearance and transmission electron microscopy revealed a typical mycoplasma cellular morphology. Molecular characterization included analysis of the following genetic loci: 16S rRNA, 23S rRNA, 16S-23S rRNA ITS, rpoB, rpoC, rpoD, uvrA, parC, topA, dnaE, fusA and pyk. The genome was sequenced for type strain 1220T. The 16S rRNA gene sequences of studied strains of Mycoplasma sp. 1220 shared 99.02-99.19 % nucleotide similarity with M. anatis strains but demonstrated ≤95.00-96.70 % nucleotide similarity to the 16S rRNA genes of other species of the genus Mycoplasma. Phylogenetic, average nucleotide and amino acid identity analyses revealed that the novel species was most closely related to Mycoplasma anatis. Based on the genetic data, we propose a novel species of the genus Mycoplasma, for which the name Mycoplasma anserisalpingitidis sp. nov. is proposed with the type strain 1220T (=ATCC BAA-2147T=NCTC 13513T=DSM 23982T). The G+C content is 26.70 mol%, genome size is 959110 bp.


Assuntos
Gansos/microbiologia , Mycoplasma/classificação , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Feminino , Genes Bacterianos , Hungria , Mycoplasma/isolamento & purificação , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Análise de Sequência de DNA
19.
Parasitol Res ; 119(4): 1423-1427, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32107621

RESUMO

We report two cases of bovine babesiosis caused by Babesia divergens in a region of central Bosnia and Herzegovina. The cases were detected in June 2017 and July 2018 from two small backyard farms. Routine clinical assessments, including physical examination and haematology, revealed lethargy, fever, anaemia, leukopenia and haemoglobinuria in the affected animals. Serum alterations included an elevation of aspartate aminotransferase and a decrease of serum phosphate or hypophosphatemia. Thrombocytopenia was detected in the first clinical case. Microscopic examination of blood smears revealed intracytoplasmic protozoan parasites from the genus Babesia. Molecular screening of both animals confirmed the presence of Babesia divergens, the causative agent of bovine babesiosis. B. divergens DNA was also detected in two engorged female Ixodes ricinus ticks removed from these animals. In addition, Mycoplasma wenyonii DNA was identified by molecular screening in the animal examined in June 2017, and in I. ricinus ticks feeding on this animal. This study provides molecular confirmation of B. divergens as a cause of piroplasmosis in cattle in South-East Europe. The detection of M. wenyonii DNA ain I. ricinus also provides the first evidence of this bacterium in ticks in Europe.


Assuntos
Babesia/genética , Babesiose/diagnóstico , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Infecções por Mycoplasma/veterinária , Mycoplasma/genética , Animais , Aspartato Aminotransferases/sangue , Babesia/isolamento & purificação , Babesiose/parasitologia , Bósnia e Herzegóvina , Bovinos , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Europa (Continente) , Fazendas , Feminino , Hipofosfatemia/sangue , Ixodes/microbiologia , Ixodes/parasitologia , Mycoplasma/isolamento & purificação , Infecções por Mycoplasma/diagnóstico , Trombocitopenia/sangue
20.
Comp Immunol Microbiol Infect Dis ; 70: 101448, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32109761

RESUMO

The presence of DNA of hemotropic mycoplasmas (hemoplasmas) was investigated for the first time in bats in Africa. Blood samples from 90 bats captured within or near human settlements in nine study areas from five states in Nigeria belonging to six genera of the families Pteropodidae, Rhinolophidae, and Molossidae were analyzed using conventional PCR protocol targeting a 391 bp part of the 16S rRNA gene. Of these, 32 samples (35 %) resulted positive. Eight nucleotide sequence types were identified, which were assigned to five genotypes showing between 93-99 % similarity with hemoplasmas from bats and/or rodents from other parts of the world, and/or Candidatus Mycoplasma haemohominis from a human patient. Network analysis showed genetic structure of hemoplasma sequences among bat species, but some sequences were shared among bats of different taxonomic groups and distant study areas. Further characterization of the samples using a protocol targeting ∼1200 bp of the 16S rRNA gene resulted in seven sequences that confirmed the results of the screening protocol. Hemoplasmas in Nigerian bats are prevalent, widely distributed and genetically diverse. The zoonotic risk to local inhabitants should not be neglected, due to the high similarity of some of the retrieved sequences with the human pathogen C. M. haemohominis.


Assuntos
Quirópteros/microbiologia , Genótipo , Habitação , Infecções por Mycoplasma/veterinária , Mycoplasma/classificação , Animais , DNA Bacteriano/genética , Variação Genética , Geografia , Mycoplasma/isolamento & purificação , Infecções por Mycoplasma/sangue , Infecções por Mycoplasma/epidemiologia , Nigéria/epidemiologia , Filogenia , Prevalência , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
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