Hemotropic mycoplasmas in bats from forest fragments, state of Paraná, southern Brazil / Detecção e caracterização de micoplasmas hemotrópicos em morcegos de fragmentos florestais, Paraná, Sul do Brasil

The order Chiroptera is the second largest group of mammals with bats being identified as reservoir of several viral zoonoses, although, little is known about their role in other groups of pathogens, including hemotropic Mycoplasma spp. To date, hemoplasma species have been found infecting several species of bats with high genetic diversity between 16S rRNA gene sequences. On this study, we aimed to identify the occurrence and characterize 16S and 23S rRNA genes of hemoplasma species in four bats species (Artibeus lituratus, Carollia perspicillata, Sturnira lilium and Sturnira tildae) from forest fragments in Paraná State, southern Brazil, using PCR-based assays. Spleen tissue samples were collected, DNA extracted and further screened by a pan hemoplasma PCR assay. All samples consistently amplified the mammal endogenous gapdh gene. One out of 15 (6.66%; 95% CI: 0.2-31%) bats tested positive for hemotropic Mycoplasma sp. by the PCR assay targeting the 16S rRNA gene. Sequencing of the 16S rRNA gene fragment from the hemoplasma-positive bat showed 99.14% identity with hemotropic Mycoplasma sp. detected in Sturnira parvidens from Belize. Sequencing of the 23S rRNA gene fragment from the hemoplasma-positive bat showed 86.17% identity with 'Candidatus Mycoplasma haemosphiggurus' detected in orange-spined hairy dwarf porcupines (Sphiggurus villosus) from Southern Brazil.(AU)

A ordem Chiroptera é considerada a segunda maior ordem de mamíferos do mundo, sendo os morcegos identificados como reservatórios de diversas zoonoses de origem viral, contudo, pouco se sabe sobre seu papel em outros grupos de patógenos, incluindo Mycoplasma spp. Até o momento, Mycoplasma sp., foi encontrado infectando várias espécies de morcegos ao redor do mundo, com alta diversidade genética entre sequências de genes 16S rRNA. O objetivo do presente estudo foi detectar a infecção por hemoplasmas em quinze morcegos insetívoros de quatro diferentes espécies (Artibeus lituratus, Carollia perspicillata, Sturnira lilium and Sturnira tildae) provenientes de fragmentos florestais dos municípios de Mandaguaçu, Maringá e Paiçandu, no Estado do Paraná, sul do Brasil. Amostras de tecido foram coletadas e o DNA extraído, para posterior análise por PCR para detecção de hemoplasmas. Todas as amostras amplificaram o gene gapdh. Um morcego, do total de 15 (6.66%; 95% CI: 0.2-31%), foi positivo para Mycoplasma sp. na análise do gene16S rRNA. O sequenciamento deste fragmento genético mostrou 99,14% de identidade com Mycoplasma sp. detectado em Sturnira parvidens em Belize. O sequenciamento do fragmento do gene 23S rRNA do morcego positivo mostrou 86,17% de identidade com 'Candidatus Mycoplasma haemosphiggurus' detectado em ouriço-cacheiro (Sphiggurus villosus) no sul do Brasil.(AU)

Detection of Hemotropic Mycoplasma sp. in white-eared opossums (Didelphis albiventris) from Southern Brazil / Detecção de Mycoplasma sp. hemotrópico em gambás-de-orelha-branca (Didelphis albiventris) no Sul do Brasil

Opossums are marsupials from the New World of the genus Didelphis and known as synanthropic animals due to their proximity with human beings. To date, Candidatus Mycoplasma haemodidelphis has been solely found infecting the North American opossum (Didelphis virginiana). Accordingly, the aim of this study was to screen eight white-eared opossums (Didelphis albiventris) from a public park in Maringa city, Paraná State, southern Brazil, for hemoplasma infection. Blood samples were taken from caudal venipuncture, and DNA was extracted and further screened by a pan-hemoplasma PCR assay. Seven out of eight (87.50%; CI 95%: 47.35-99.68%) white-eared opossums were positive for Mycoplasma spp. Sequencing of the 16S rRNA fragment showed 98,97% identity with Ca. M. haemodidelphis detected in the USA. Three out of eight (37.50%; CI 95%: 8.52-75.51%) white-eared opossums were infested by Amblyomma dubitatum ticks. This is the first report on detection of a potentially novel hemotropic Mycoplasma sp. infecting opossums from South America.(AU)

Gambás são marsupiais do Novo Mundo, pertencentes ao gênero Didelphis, e considerados animais sinantrópicos devido à sua proximidade com seres humanos. Atualmente, a espécie Candidatus Mycoplasma haemodidelphis só foi encontrada infectando gambá norte americano (Didelphis virginiana). O objetivo do presente estudo foi detectar a infecção por hemoplasmas em oito gambás-de-orelha-branca (Didelphis albiventris) capturados em um parque público da cidade de Maringá, no Estado do Paraná, sul do Brasil. Amostras de sangue foram coletadas por venopunção caudal para a extração do DNA e posterior análise pela PCR para espécies de hemoplasmas. Sete de oito animais (87,50%; CI 95%: 47,35-99,68%) foram considerados positivos para Mycoplasma spp. O sequenciamento do fragmento do gene 16S rRNA obtido apresentou 98.97% de similaridade com sequências de Ca. M. haemodidelphis detectadas nos Estados Unidos. Três gambás (37,50%; CI 95%: 8,52-75,51%) estavam infestados por carrapatos da espécie Amblyomma dubitatum. Esse é o primeiro relato de detecção de uma potencial nova espécie de Mycoplasma hemotrópico infectando gambás na América do Sul.(AU)

Screening of feral pigeons (columba livia) for pathogens of veterinary and medical importance

Pathogens of veterinary and medical importance were investigated in 240 feral pigeons (Columba livia) captured in warehouses in São Paulo State, Brazil for one year. Rapid serum agglutination test (RST) was performed for the detection of antibodies against Mycoplasma synoviae, Mycoplasma gallisepticum and Salmonella Pullorum/Gallinarum. Positive samples were submitted to hemagglutination inhibition (HI) and tube seroagglutination tests, respectively. Molecular techniques (RT-PCR and PCR) were performed for Newcastle Diseases Virus (NDV) and Chlamydia psittaci diagnosis. Additionally, HI test was applied to detect antibodies against NDV. Serological results by RST were 3.3% positive for M. synoviae, 2.5% for M. gallisepticum, and 0.4% for S. Pullorum/Gallinarum, all negative on the confirmatory tests performed. NDV RNA or antibodies were not detected. C. psittaci DNA was detected in 13% of the samples. Further research on pigeon health status should be conducted because this species is highly adaptable and their numbers are rapidly rising around the world, posing risks for animals and human beings.

Screening of feral pigeons (columba livia) for pathogens of veterinary and medical importance

Pathogens of veterinary and medical importance were investigated in 240 feral pigeons (Columba livia) captured in warehouses in São Paulo State, Brazil for one year. Rapid serum agglutination test (RST) was performed for the detection of antibodies against Mycoplasma synoviae, Mycoplasma gallisepticum and Salmonella Pullorum/Gallinarum. Positive samples were submitted to hemagglutination inhibition (HI) and tube seroagglutination tests, respectively. Molecular techniques (RT-PCR and PCR) were performed for Newcastle Diseases Virus (NDV) and Chlamydia psittaci diagnosis. Additionally, HI test was applied to detect antibodies against NDV. Serological results by RST were 3.3% positive for M. synoviae, 2.5% for M. gallisepticum, and 0.4% for S. Pullorum/Gallinarum, all negative on the confirmatory tests performed. NDV RNA or antibodies were not detected. C. psittaci DNA was detected in 13% of the samples. Further research on pigeon health status should be conducted because this species is highly adaptable and their numbers are rapidly rising around the world, posing risks for animals and human beings.(AU)

DETECÇÃO DE MICOPLASMAS HEMOTRÓPICOS EM GAMBÁS (DIDELPHIS ALBIVENTRIS) E MORCEGOS (ARTIBEUS LITURATUS) DE FRAGMENTOS FLORESTAIS DA REGIÃO NORTE DO PARANÁ, BRASIL

Os micoplasmas hemotrópicos (hemoplasmas), bactérias gram negativas eritrocíticas e pleomórficas são responsáveis por causar uma anemia hemolítica imunomediada tanto em animais quanto em humanos. A análise da sequência do gene 16S rRNA e 23S rRNA tem impulsionado o estudo de hemoplasmas. Os gambás de orelha branca (Didelphis albiventris) assim como os morcegos, ordem Chiroptera, são importantes representantes da relação entre homem-ambiente pois são de fácil adaptação e distribuídos nas áreas urbanas e rurais. Estudos com estas espécies são importantes uma vez que são reservatórios competentes de diferentes patógenos transmitidos por vetores (DTV) e hemoplasmas, além de servirem como sentinelas para diferentes doenças tropicais. No Brasil, apenas um gambá de orelha branca foi avaliado e este foi negativo para Mycoplasma sp. por meio da PCR. Em morcegos Mycoplasma sp. foi detectado tanto em animais hematófagos quanto em não-hematófagos provenientes de cinco estados brasileiros (Mato Grosso, Pará, Paraná, São Paulo e Tocantins) e algumas sequências detectadas foram correlacionadas a Mycoplasma sp. provenientes de morcegos da América Central e do Sul. Neste contexto, o objetivo do presente estudo foi avaliar a presença de Mycoplasma sp. em gambás de orelha branca (Didelphis albiventris) e em morcegos (Chiropteros) provenientes de fragmentos florestais da região norte do Estado do Paraná, Sul do Brasil. Sete de oito gambás de orelha branca (87.50%; CI 95%: 47, 35-99, 68%) foram considerados positivos para Mycoplasma spp. O sequenciamento do fragmento do gene 16S rRNA obtido apresentou 98.97% de similaridade com sequências de Candidatus Mycoplasma haemodidelphis (AF178676) detectadas até o momento apenas nos Estados Unidos. Três gambás (37.50%; CI 95%: 8.52-75, 51%) estavam infestados por carrapatos da espécie Amblyomma dubitatum. Esse foi o primeiro relato de detecção de uma potencial nova espécie de micoplasma hemotrópico infectando gambás na América do Sul. Três espécies de morcegos foram identificadas, Artibeus lituratus (7/11, 63%), Carollia perspicillata (2/11, 18%) e Sturnira tildae (2/11, 18%). Apenas um/11 (0.9%; IC 95%) morcego (A. lituratus) foi positivo para Mycoplasma sp. O sequenciamento do fragmento do gene 16S rRNA obtido apresentou 99% de identidade com Mycoplasma sp. identificado em estudos prévios em Desmodus rotundus na América Central. Os resultados obtidos sugerem que esta potencial nova espécie de hemoplasma infecte morcegos hematófagos e não hematófagos na América Central e do Sul. Na análise do fragmento do gene 23S rRNA o Mycoplasma sp. detectado no presente estudo formou um clado com Mycoplasma sp. detectado em porcos espinhos (Sphiggurus villosus) provenientes do estado do Paraná.

Hemotropic mycoplasmas (hemoplasmas), erythrocytic and pleomorphic gram-negative bacteria are responsible for causing immune-mediated hemolytic anemia in both animals and humans. The sequence analysis of the 16S rRNA and 23S rRNA gene has driven the study of hemoplasms. White-eared possums (Didelphis albiventris) as well as bats, order Chiroptera, are important representatives of the relationship between man and the environment as they are easy to adapt and distributed in urban and rural areas. Studies with these species are important since they are competent reservoirs of different pathogens transmitted by vectors (DTV) and hemoplasms, in addition to serving as sentinels for different tropical diseases. In Brazil, only one white-eared possum was evaluated and it was negative for Mycoplasma sp. through PCR. In bats Mycoplasma sp. was detected in both hematophagous and non-hematophagous animals from five Brazilian states (Mato Grosso, Pará, Paraná, São Paulo and Tocantins) and some detected sequences were correlated to Mycoplasma sp. from bats in Central and South America. In this context, the objective of the present study was to evaluate the presence of Mycoplasma sp. in white-eared possums and in bats (Chiropteros) from forest fragments in the northern region of the State of Paraná, southern Brazil. Seven out of eight white-eared possums (87,50%; CI 95%: 47, 35-99, 68%) were considered positive for Mycoplasma spp. The sequencing of the fragment of the 16S rRNA gene obtained showed 98,97% similarity with 'Candidatus Mycoplasma haemodidelphis' (AF178676) sequences detected so far only in the United States. Three possums (37,50 %; CI 95%: 8,52-75,51%) were infested with ticks of the species Amblyomma dubitatum. This was the first report of detection of a potential new species of hemotropic mycoplasma infecting opossums in South America. Three species of bats were identified, Artibeus lituratus (7/11: 63%), Carollia perspicillata (2/11: 18%) and Sturnira tildae (2/11:18%). Only one / 11 (0,9%; CI 95%) bat (A. lituratus) was positive for Mycoplasma sp. The sequencing of the fragment of the 16S rRNA gene obtained showed 99% identity with Mycoplasma sp. identified in previous studies on Desmodus rotundus in Central America. The results obtained suggest that this potential new species of hemoplasma infects hematophagous and non-hematophagous bats in Central and South America. In the analysis of the 23S rRNA gene fragment, Mycoplasma sp. detected in the present study formed a clade with Mycoplasma sp. detected in porcupines (Sphiggurus villosus) from the state of Paraná.