Hemoplasmas in wild rodents and marsupials from the Caatinga Biome, Brazil.

A total of 231 blood samples from wild mammals belonging to the orders Rodentia (n = 142) and Didelphimorphia (n = 89) were screened by real-time PCR assay (qPCR), being six Rhipidomys sp., 118 Thrichomys laurentius, nine Rattus rattus, four Kerodon rupestris, five Necromys lasiurus, 42 Didelphis albiventris and 47 Monodelphis domestica. Results using qPCR showed that 32 of the total 231 (13.85 %) samples were positive for hemoplasma sequences of the 16S rRNA gene. Sequences from two D. albiventris showed 99.77-99.89 % identity with 'Candidatus Mycoplasma haemoalbiventris' and 99.09 % with 'Candidatus Mycoplasma haemodidelphidis', respectively. Furthermore, one M. domestica and five T. laurentius showed 99.72-99.77 % identity with Mycoplasma sp., and one K. rupestris showed 98.13 % identity with 'Candidatus Mycoplasma haematohydrochaerus'; and from two Rattus rattus showed 99.65-99.89 % identity with Mycoplasma sp. and 'Candidatus Mycoplasma haemomuris'. The 23S rRNA gene sequences obtained from the two D. albiventris showed 100 % identity with 'Ca. M. haemoalbiventris' whereas the sequences from the R. rattus showed only 85.31 % identity with 'Candidatus Mycoplasma haematohydrochaerus'. Two T. laurentius and one K. rupestris showed 84.66-92.97 % identity with 'Candidatus Mycoplasma haemosphiggurus'. Based on phylogenetic and Neighbor-Net network analyses of the 16S and 23S rRNA genes, potential novel species are described. In addition, 'Ca. M. haemoalbiventris' was detected in Didelphis albiventris, and Mycoplasma sp. was detected in Rattus sp. rodents from the Caatinga biome, Brazil.

'Candidatus Mycoplasma Haemoalbiventris' and Tick-Borne Pathogens in Black-Eared Opossum (Didelphis aurita) from Southeastern Brazil.

The black-eared opossum (Didelphis aurita) is a South American synanthropic marsupial. The presence of opossums in domestic spaces is relevant in the One-Health context since they are hosts of pathogens and ectoparasites that may affect the health of domestic animals and humans. In this study, we aim to determine the occurrence of hemoplasmas and selected tick-borne pathogens in free-ranging black-eared opossums, along with their molecular characterization, hematological and biochemical evaluation and factors associated with infection, in the municipality of Viçosa, State of Minas Gerais, southeastern Brazil. Thirty black-eared opossums were trapped between March 2021 and June 2022. Ectoparasites were collected. Hematological and biochemical analyses were performed. DNA from EDTA-blood samples were analyzed by PCR and qPCR assays. By molecular analyses, 'Candidatus Mycoplasma haemoalbiventris' was the most prevalent hemoparasite (73.3%), followed by Hepatozoon sp. (22.2%). Significant differences were observed in the number of platelets, and in the concentration of protein and globulins in the animals infected by 'Ca. M. haemoalbiventris' when compared with the negative group. This is the first report of 'Ca. M. haemoalbiventris' infection in D. aurita.

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)

Molecular screening of Bartonella in free-ranging capybaras (Hydrochoerus hydrochaeris) from Paraná State, Southern Brazil / Avaliação molecular de Bartonella em capivaras (Hydrochoerus hydrochaeris) de vida livre do Estado do Paraná, Brasil

Bartonella is an emerging group of facultative intracellular bacteria causing circulatory and systemic disorders. Hosts for Bartonella are mostly mammals, specifically rodents, having a growing number of Bartonella species related to their infection. Capybaras (Hydrochoerus hydrochaeris) are abundant native rodents of Brazil, commonly found in urban parks. In the present study, we aimed to perform molecular screening of capybaras for Bartonella spp. Blood samples were collected from 17 free-ranging animals captured in Paraná State, Southern Brazil. None of the collected samples tested positive for the BartonellanuoG gene by quantitative polymerase chain reaction (qPCR), although all of them successfully amplified the mammal endogenous glyceraldehyde-3-phosphate dehydrogenase (gapdh) gene. Additionally, all animals were infested exclusively by Amblyomma dubitatum ticks at the time of sampling. This study was part of an active surveillance program, which is critical for monitoring animal health status, particularly in capybaras.(AU)

Bartonella é um grupo emergente de bactérias intracelulares facultativas que causam doenças circulatórias e sistêmicas. Mamíferos são os principais hospedeiros das bartonelas, especificamente roedores, com crescente número de espécies de Bartonella relacionadas a estes animais. Capivaras (Hydrochoerus hydrochaeris) são abundantes roedores do Brasil, comumente encontrados em parques urbanos. Nosso objetivo no presente estudo foi realizar uma triagem molecular das capivaras para Bartonella spp. Amostras de sangue foram coletadas de 17 animais de vida livre capturados no estado do Paraná, Sul do Brasil. Nenhuma das amostras coletadas apresentou resultado positivo para o gene nuoG de Bartonella pela reação em cadeia da polimerase quantitativa (qPCR), apesar de todas elas amplificarem com sucesso o gene endógeno de mamíferos gliceraldeído-3-fosfato desidrogenase (gapdh). Adicionalmente, todos os animais estavam infestados exclusivamente por carrapatos Amblyomma dubitatum no momento da coleta. Este estudo foi parte do programa de vigilância ativa, que é importante para monitorar a condição de saúde dos animais, particularmente em capivaras.(AU)

'Candidatus Mycoplasma haemoalbiventris' and tick-borne pathogens screening in white-eared opossums (Didelphis albiventris) from Curitiba and Foz do Iguaçu Cities, Paraná State, southern Brazil.

Hemoplasmas are epierythrocytic bacteria that infect mammals. 'Candidatus Mycoplasma haemoalbiventris' was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by 'Ca. M. haemoalbiventris'. Although 'Ca. M. haemoalbiventris' is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.

‘Candidatus Mycoplasma haemoalbiventris and tick-borne pathogens screening in white-eared opossums (Didelphis albiventris) from Curitiba and Foz do Iguaçu Cities, Paraná State, southern Brazil / ‘Candidatus Mycoplasma haemoalbiventris e triagem para patógenos transmitidos por carrapatos em gambás de orelha branca (Didelphis albiventris) das cidades de Curitiba e Foz do Iguaçu, Estado do Paraná, sul do Brasil

Hemoplasmas are epierythrocytic bacteria that infect mammals. ‘Candidatus Mycoplasma haemoalbiventris was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by ‘Ca. M. haemoalbiventris. Although ‘Ca. M. haemoalbiventris is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.(AU)

Hemoplasmas são bactérias epieritrocíticas que infectam mamíferos. ‘Candidatus Mycoplasma haemoalbiventris foi detectado previamente em gambás-de-orelha-branca (Didelphis albiventris) das regiões sul e centro-oeste do Brasil. O presente estudo objetivou: i) triar os gambás para as doenças transmitidas por carrapatos (Piroplasmida e Anaplasmataceae); e ii) detectar e caracterizar as espécies de hemoplasma que infectam gambás nas cidades de Curitiba e Foz do Iguaçu, no Estado do Paraná, sul do Brasil. Trinta amostras de sangue de gambás-de-orelha-branca foram analisadas por PCR. Os animais não estavam infestados por ectoparasitos. O gene endógeno de mamífero gapdh foi amplificado em todas as amostras. Todos os gambás testaram negativos para Theileria/Babesia spp. e Ehrlichia/Anaplasma spp. por PCR, respectivamente, para os genes 18S rRNA e 16S rRNA. Uma PCR gene-específica, baseada no gene 16S rRNA de hemoplasmas, mostrou que três/13 (23,08%; CI 95%: 8,18-50,26%) gambás de Foz do Iguaçu foram positivos para Mycoplasma sp. hemotrópico. Todos os gambás de Curitiba testaram negativos para hemoplasmas. O sequenciamento de fragmentos dos genes 16S e 23S rRNA revelou que os animais estavam infectados pelo ‘Ca. M. haemoalbiventris. Embora ‘Ca. M. haemoalbiventris seja prevalente em gambás no Brasil, os sinais clínicos associados à infecção e os prováveis vetores permanecem desconhecidos.(AU)

'Candidatus Mycoplasma haemoalbiventris' and tick-borne pathogens screening in white-eared opossums (Didelphis albiventris) from Curitiba and Foz do Iguaçu Cities, Paraná State, southern Brazil / 'Candidatus Mycoplasma haemoalbiventris' e triagem para patógenos transmitidos por carrapatos em gambás de orelha branca (Didelphis albiventris) das cidades de Curitiba e Foz do Iguaçu, Estado do Paraná, sul do Brasil

Abstract Hemoplasmas are epierythrocytic bacteria that infect mammals. 'Candidatus Mycoplasma haemoalbiventris' was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by 'Ca. M. haemoalbiventris'. Although 'Ca. M. haemoalbiventris' is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.

Resumo Hemoplasmas são bactérias epieritrocíticas que infectam mamíferos. 'Candidatus Mycoplasma haemoalbiventris' foi detectado previamente em gambás-de-orelha-branca (Didelphis albiventris) das regiões sul e centro-oeste do Brasil. O presente estudo objetivou: i) triar os gambás para as doenças transmitidas por carrapatos (Piroplasmida e Anaplasmataceae); e ii) detectar e caracterizar as espécies de hemoplasma que infectam gambás nas cidades de Curitiba e Foz do Iguaçu, no Estado do Paraná, sul do Brasil. Trinta amostras de sangue de gambás-de-orelha-branca foram analisadas por PCR. Os animais não estavam infestados por ectoparasitos. O gene endógeno de mamífero gapdh foi amplificado em todas as amostras. Todos os gambás testaram negativos para Theileria/Babesia spp. e Ehrlichia/Anaplasma spp. por PCR, respectivamente, para os genes 18S rRNA e 16S rRNA. Uma PCR gene-específica, baseada no gene 16S rRNA de hemoplasmas, mostrou que três/13 (23,08%; CI 95%: 8,18-50,26%) gambás de Foz do Iguaçu foram positivos para Mycoplasma sp. hemotrópico. Todos os gambás de Curitiba testaram negativos para hemoplasmas. O sequenciamento de fragmentos dos genes 16S e 23S rRNA revelou que os animais estavam infectados pelo 'Ca. M. haemoalbiventris'. Embora 'Ca. M. haemoalbiventris' seja prevalente em gambás no Brasil, os sinais clínicos associados à infecção e os prováveis vetores permanecem desconhecidos.