First molecular detection of Mycoplasma ovis in horses from Brazil.

This study aimed to determine the occurrence of hemoplasmas and tick-borne pathogens (TBP) (Theileria equi, Babesia caballi, and Ehrlichia sp.) in horses and ticks' salivary glands, and determine the factors associated with exposure/infection in a rural settlement in southern Brazil. Blood samples from 22 horses were screened for anti-T. equi and anti-Ehrlichia sp. antibodies by an indirect fluorescent antibody test (IFAT) assays. Samples were also tested by PCR assays for T. equi and B. caballi (18S rRNA and rap-1 genes, respectively), hemoplasmas (16S rRNA gene), and Ehrlichia sp. (dsb gene). Ticks were removed from the animals (inspection) and the environment (flannel trawling and dry ice traps), and morphologically identified. Additionally, salivary glands DNA was extracted from 28 adult ticks infesting the animals and four nymphs from the environment, and further screened for Ehrlichia sp. and hemotropic Mycoplasma sp. Anti-T. equi and anti-Ehrlichia sp. antibodies were detected in 40.91% (nine/22; 95% CI: 23.26-61.27) and 31.81% (seven/22; 95% CI: 16.36-52.68) horses, respectively. Theileria equi, B. caballi, and hemotropic Mycoplasma sp. DNA was detected in 59.09% (13/22), 4.55% (one/22), and 50% (11/22) horses, respectively. All horses tested negative in the PCR for Ehrlichia sp. All sequences showed ≥99% identity with multiple T. equi, B. caballi, and Mycoplasma ovis sequences deposited in GenBank database. Adult ticks were identified as Dermacentor nitens (44/47; 93.62%) and Rhipicephalus microplus (three/47; 6.38%). Ticks' salivary glands were negative for Ehrlichia sp., while 39.29% from adults (11/28) and 50% from nymphs (two/four) from the environment were positive for hemotropic Mycoplasma sp. This is the first report of M. ovis infection in horses from Brazil and the first detection of hemoplasma DNA in salivary glands of D. nitens and R. microplus ticks. Further studies are needed to elucidate the vector competence of ticks to transmit hemoplasmas.

Hemoplasmas and ticks in cattle from Somalia.

Hemotropic mycoplasmas (hemoplasmas) are small Gram-negative bacteria that parasitize red blood cells and can cause mild to severe anemia in a wide range of vertebrates, including ruminants. Cattle population in Somalia is around 3.9 million heads, with animals more concentrated around the river areas, mainly in the Juba River and Shabelle River Valleys. Information on hemoplasmas in Sub-Saharan Africa are scarce, mainly in Somalia, where no studies have been performed to date. Accordingly, this study aimed to assess the molecular occurrence of hemoplasmas in 131 cattle blood samples from Somalia. Thirty out of 131 (22.90%; 95% CI: 16.54-30.81%) cattle were infested by ticks: Rhipicephalus pulchellus (68.18%), Amblyomma gemma (18.18%), Amblyomma lepidum (9.09%), Hyalomma marginatum (1.51%), Hyalmomma rufipes (1.51%), and Rhipicephalus pravus (1.51%). A total of 74/131 (56.48%; 95% CI: 47.93-64.67%) cattle were positive for hemotropic Mycoplasma spp. by real-time PCR (qPCR) based on the 16S rRNA gene. Hemoplasma-positive samples were later subjected to species-specific PCR assays for Mycoplasma wenyonii and 'Candidatus Mycoplasma haematobovis' based on the 16S rRNA gene. A total of 34/74 (45.94%; 95% CI: 35.07-57.22%) animals were coinfected by both species; 31/74 (41.89%; 95% CI: 31.32-53.26%) and 3/74 (4.05%; 95% CI: 01.39-11.25%) cattle were solely positive to M. wenyonii and 'Ca. M. haematobovis', respectively. Six out of 74 (8.1%; 95% CI: 03.77-16.58%) cattle were negative on species-specific conventional PCR assays but tested positive by a semi-nested PCR assay based on the 16S rRNA gene of hemoplasmas. Sequencing of the detected hemotropic Mycoplasma sp. 16S rRNA gene confirmed that animals were infected by M. wenyonii and 'Ca. M. haematobovis'. To the best of our knowledge, this is the first study on the detection of hemoplasmas in cattle from Somalia.

Novel Anaplasmataceae agents Candidatus Ehrlichia hydrochoerus and Anaplasma spp. Infecting Capybaras, Brazil.

We amplified Ehrlichia and Anaplasma DNA from Amblyomma dubitatum tick-infested capybaras (Hydrochoerus hydrochaeris) in southern Brazil. Sequencing of 16S rRNA, sodB, and groEL indicated a novel Ehrlichia species, and sequencing of 16S rRNA from 2 capybaras indicated a novel Anaplasma species. The tick vectors remain unknown.

'Candidatus Mycoplasma haematonasua' and tick-borne pathogens in ring-tailed coatis (Nasua nasua Linnaeus, 1976) from the Iguaçu National Park, Paraná State, southern Brazil.

The Iguaçu National Park (INP) is the largest remnant of Atlantic Forest in southern Brazil, representing an ecological continuum with Argentina. The INP harbours a diverse fauna, with ring-tailed coatis (Nasua nasua Linnaeus, 1976, Carnivora: Procyonidae) in close contact with tourists either begging and/or snatching food from visitors. A potentially novel haemotropic Mycoplasma sp. has been previously detected in the ring-tailed coatis from central-western and southern Brazil. Therefore, the aims of this study were to investigate the occurrence of haemotropic Mycoplasma sp. and tick-borne pathogens in wild ring-tailed coatis from the INP, Foz do Iguaçu municipality, Paraná State, southern Brazil. Blood samples were collected from 18 wild ring-tailed coatis and evaluated by conventional PCR (cPCR) assays for haemotropic Mycoplasma spp. (16S and 23S rRNA), Theileria/Babesia spp. (18S rRNA) and Ehrlichia/Anaplasma spp. (16S rRNA, sodB, dsb and groEL). Eight out of 18 (44.44%; 95% CI: 24.56%-66.28%) animals were positive for haemotropic Mycoplasma spp. All ring-tailed coatis tested negative for Theileria/Babesia spp. and only one out of 18 (5.56%; 95% CI: 0.99%-25.76%) animals tested positive for Ehrlichia/Anaplasma spp. by the 16S rRNA cPCR. Unfortunately, multiple attempts to sequence the 16S rRNA gene of the Ehrlichia/Anaplasma-positive sample have failed. Phylogenetic and network analysis of the hemoplasma 16S and 23S rRNA gene fragments confirmed that animals were infected by a potentially novel haemotropic Mycoplasma sp. previously reported in ring-tailed coatis from Brazil. The name 'Candidatus Mycoplasma haematonasua' is proposed for this novel organism.

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

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