Post-mortem detection of hemoplasmas (hemotropic Mycoplasma spp.) in South American fur seal (Arctocephalus australis) sampled in Rio Grande do Sul State, southern Brazil.

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 16 S rRNA and 23 S rRNA gene. Three (2.22 %) Arctocephalus australis were positive in the 16 S rRNA gene, and no samples amplified in the 23 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.

Description of Amblyomma monteiroae n. sp. (Acari: Ixodidae), a parasite of the great horned owl (Strigiformes: Strigidae) in southern Brazil.

In 2020, adult hard ticks (males and females) were collected from great horned owls [Bubo virginianus (Gmelin, 1788)] in the coastal region in southern Brazil. The engorged females were allowed to oviposit in the laboratory and hatched larvae could be obtained. Analyses of the external morphology of the adult ticks revealed that they represent a new species, which was named Amblyomma monteiroae n. sp. Partial sequences of the mitochondrial 16S rRNA gene and the nuclear second internal transcribed spacer (ITS2) were generated from a male and a female. Their 16S rRNA haplotypes were identical to each other and closest (96% identity) to corresponding sequences of Amblyomma parvitarsum Neumann, 1901, and 90% identical to Amblyomma neumanni Ribaga, 1902. Their ITS2 haplotypes were 95.8 to 96.0 identical to the single ITS-2 partial sequence of A. parvitarsum available in GenBank. In the phylogenetic trees inferred by both 16S rRNA and ITS2 partial sequences, A. monteiroae n. sp. formed a clade with A. parvitarsum, with A. neumanni branching sister to this clade. Amblyomma monteiroae n. sp. is genetically and morphologically related to A. parvitarsum. Both tick species are unique in combining the following morphological characters: scutum extensively ornate; eyes rounded and bulging; coxa I with two moderate pointed spurs, the external longer than the internal; a single triangular short spur on coxae II-III; presence of two spines on the tibia of legs II-IV; hypostomal dentition 3/3, trochanters without spurs. However, the males of the two species can be separated by specific features in palps and festoons, whereas the females differ in specific features of the coxal spurs. The larva of A. monteiroae n. sp. can be morphologically distinguished from A. parvitarsum only by morphometry, with the former species being slightly smaller. Currently, A. monteiroae n. sp. is restricted to southern Brazil, and the only known host is B. virginianus (Strigiformes: Strigidae). The present study increases the Amblyomma Brazilian fauna to 34 species.

Molecular survey of hemotropic mycoplasmas in crab-eating raccoons (Procyon cancrivorus) in southern Brazil.

Hemoplasmas are non-cultivable bacterial parasites of erythrocytes that infect domestic and wild animals, as well as humans. Their means of transmission and pathogenesis remain contentious issues and difficult to evaluate in wild animals. Procyon cancrivorus is a South American carnivore and occurs in all Brazilian biomes. In this study, we aimed to investigate occurrences of hemoplasmas infecting P. cancrivorus and to identify their 16S rRNA gene, in southern Brazil. DNA was extracted from spleen and blood samples of P. cancrivorus (n = 9) from different locations. Hemoplasma DNA was detected in six samples, based on 16S rRNA gene amplification and phylogenetic analysis. Four of the six sequences belonged to the "Mycoplasma haemofelis group", which is closely related to genotypes detected in Procyon lotor from the USA; one was within the "Mycoplasma suis group", closely related to "Candidatus Mycoplasma haemominutum"; and one was within the intermediate group between these clusters. Thus, these sequences showed that the molecular identity of hemoplasmas in the population studied was very variable. In five positive animals, Amblyomma aureolatum ticks and a flea (Ctenocephalides felis felis) were collected. The present study describes the first molecular detection of mycoplasmas in P. cancrivorus.

Epidemiological compatibility of Amblyomma sculptum as possible vector and Panthera onca as reservoir of Cytauxzoon spp. in Midwestern Brazil.

Cytauxzoonosis is an acute and highly lethal tick-borne disease of wild and domestic cats, and is widely distributed in Africa, Asia, Europe, the USA and Brazil. So far, only two tick species present on the USA are experimentally confirmed in Cytauxzoon transmission however, in Brazil and other continents, the epidemiology of the disease remains unknown. Evidences points to Panthera onca as a possible reservoir, but there is no evidence to point the vector. Therefore, this study evaluates the presence of Cytauxzoon spp. in wild felids from areas with and without records of Amblyomma sculptum this ixodid for comparison. Overall, 53 blood samples of P. onca, Puma concolor, and Leopardus pardalis from the Midwest region (MR; region with A. sculptum) and 143 blood and/or spleen samples from Leopardus geoffroyi, Leopardus wiedii, Leopardus munoai, Leopardus guttulus, Herpailurus yagouaroundi, L. pardalis, and P. concolor from Rio Grande do Sul State (RS; without A. sculptum). Only one feline sample was negative for Cytauxzoon sp. from MR; no samples from RS were positive. In total, 507 ticks were identified from MR felids, with predominance of A. sculptum (69.23%). In RS, there were 93 ixodids, of which 90.32% were Amblyomma aureolatum. The difference in the tick fauna of the two regions studied (presence/absence of A. sculptum) reflects the results found. This study highlighted A. sculptum as a possible vector since this hemoparasite was abundantly observed in areas where it occurs, also, there was no evidence of Cytauxzoon spp. where it was absent. Additionally, the study supported the suggestion that P. onca is the reservoir for the agent in MR.

Rangelia vitalii molecular and histological quantification in tissues comparing crab-eating foxes (Cerdocyon thous) and domestic dogs.

Rangeliosis is a condition transmitted by the tick Amblyomma aureolatum and caused by the protozoan parasite Rangelia vitalii in canids. In domestic dogs, the disease causes a severe hemolytic disease, while in wild canids the piroplasm is often detected without any clinical abnormality. This study aimed to detect and quantify the number of copies of the R. vitalii Hsp70 gene (indirect parasite burden) in several organs of domestic and South American wild canids (Cerdocyon thous and Lycalopex gymnocercus) to elucidate distinct clinical presentations of rangeliosis in these species. A total of seven domestic dogs that died due to rangeliosis and 38 wild foxes were initially included, with all dogs presenting histological and molecular features of rangeliosis, while eight C. thous were positive at the molecular analysis for R. vitalii. Fragments of 22 organs collected from domestic (n = 7) and wild foxes (n = 8) were employed for histological and molecular quantification using real-time polymerase chain reaction aiming at the Hsp70 gene. Histologically, parasitophorous vacuoles were constantly detected in the dogs, while these were detected only in two C. thous. Parasitic burden was significantly higher in the digestive, cardiorespiratory, endocrine, genitourinary, and skeletal-muscle systems of domestic dogs when compared to wild foxes. In the hematopoietic system of wild canids, some organs, such as the lymph nodes and tonsils, presented significantly lower amounts of R. vitalii, while other organs (spleen, bone marrow, and blood) had results similar to those of domestic dogs. Additionally, the central nervous system of both domestic and wild canids presented a similar quantity of R. vitalii. The etiological agent is possibly maintained through an asexual reproductive process (merogony) in both domestic and wild species. Nonetheless, a limited or short-duration schizogony phase occurs in C. thous, which would designate this species as a possible reservoir host for the agent. Dogs, in contrast, would most likely act as accidental hosts, presenting a severe and more pathogenic schizogony phase, resulting in characteristic clinical and pathological rangeliosis.

Slaughter condemnation in bovine due to parasitic lesions and their economic impact in Federal Inspection System establishments in Brazil and in State inspection System in Rio Grande do Sul State.

Slaughter condemnations are important sources of information on cattle health. The incidence of bovine parasitic diseases is still very high in Brazil. These diseases, in addition to causing harm to the animals health, are neglected zoonotic diseases in several parts of world. The study analysed not only the Carcass losses, but also the economic damage resulting from slaughter condemnations due to parasitic causes. Cattle slaughter data from the Brazilian Ministry of Agriculture, Livestock and Supply (MAPA), over the period of 2012 to 2015 and data from the Secretary of Livestock and Irrigation of the state of Rio Grande do Sul (SEAPI-RS) were analyzed between 2014 and 2018.The number of organs and carcasses condemned was multiplied by the respective values (in Brazilian Real) obtained from slaughterhouses and subsequently converted into dollars. Brazilian analysis in SIF (Federal Inspection System) establishments showed that more than 1.2 million organs (3,884,505 kg) and 20,000 carcasses (4,547,718 kg) were condemned only due to parasitic causes during post-mortem inspection. In Rio Grande do Sul, in state inspection establishments, more than 1.7 million organs (8,210,559 kg) and 5,000 carcasses (1,243,200 kg) were condemned. These data are alarming and support the need for public policies to control these parasitic diseases.

Slaughter condemnation in bovine due to parasitic lesions and their economic impact in Federal Inspection System establishments in Brazil and in State inspection System in Rio Grande do Sul State / Condenação ao abate em bovinos devido a lesões parasitárias e seu impacto econômico em estabelecimentos do Sistema Federal de Inspeção no Brasil e no Sistema Estadual de Inspeção no Rio Grande do Sul

Abstract Slaughter condemnations are important sources of information on cattle health. The incidence of bovine parasitic diseases is still very high in Brazil. These diseases, in addition to causing harm to the animals health, are neglected zoonotic diseases in several parts of world. The study analysed not only the Carcass losses, but also the economic damage resulting from slaughter condemnations due to parasitic causes. Cattle slaughter data from the Brazilian Ministry of Agriculture, Livestock and Supply (MAPA), over the period of 2012 to 2015 and data from the Secretary of Livestock and Irrigation of the state of Rio Grande do Sul (SEAPI-RS) were analyzed between 2014 and 2018.The number of organs and carcasses condemned was multiplied by the respective values (in Brazilian Real) obtained from slaughterhouses and subsequently converted into dollars. Brazilian analysis in SIF (Federal Inspection System) establishments showed that more than 1.2 million organs (3,884,505 kg) and 20,000 carcasses (4,547,718 kg) were condemned only due to parasitic causes during post-mortem inspection. In Rio Grande do Sul, in state inspection establishments, more than 1.7 million organs (8,210,559 kg) and 5,000 carcasses (1,243,200 kg) were condemned. These data are alarming and support the need for public policies to control these parasitic diseases.

Resumo Condenações durante o abate são importantes fontes de informação sobre a sanidade dos rebanhos. Ainda são altas as incidências de doenças parasitárias, as quais trazem consequências à saúde dos animais, são zoonoses e têm sua importância negligenciada em diversos países. O estudo objetivou quantificar não apenas as perdas encontradas, mas o prejuízo econômico resultante das condenações ao abate por causas parasitárias. Para isso foram analisados dados de abate de bovinos do Ministério da Agricultura Pecuária e Abastecimento, no período de 2012 a 2015, e dados da Secretaria da Agricultura Pecuária e Irrigação, do estado do Rio Grande do Sul, no período de 2014 a 2018. A quantidade de órgãos e carcaças condenados foi multiplicada pelos respectivos valores (em Reais), obtidos da indústria frigorífica e, posteriormente, convertidos para dólar. A análise brasileira de estabelecimentos com SIF (Serviço de Inspeção Federal) mostrou que mais de 1,2 milhão de órgãos (3.884.505 kg) e 20 mil carcaças (4.547.718 kg) foram condenados devido a causas parasitárias durante a inspeção. No estado do Rio Grande do Sul, nos estabelecimentos de inspeção estadual, foram condenados mais de 1,7 milhão de órgãos (8.210.559 kg) e 5 mil carcaças (1.243.200 kg). Os dados são alarmantes e ressaltam a necessidade de políticas públicas para controlar estas parasitoses.

First molecular detection of Mycoplasma ovis (Hemotropic mycoplasmas) from Sheep in Brazil / Primeira detecção molecular de Mycoplasma ovis (Micoplasmas Hemotrópicos) em Ovinos no Brasil

Abstract Mycoplasma ovis is an emerging zoonotic pathogen with a worldwide distribution and can cause mild to severe hemolytic anemia, icterus, and poor weight gain in animals. Although M. ovis has been described in small ruminants worldwide, data on M. ovis in sheep in Brazil is unknown. The objective of the present study was to present the first report of hemotropic mycoplasma (HM) in sheep from Brazil. We evaluated factors associated with this infection, such age group, tick presence, and anemia. Blood samples were collected from 33 sheep from a farm in southern Brazil and screened for hemoplasmas using PCR. Out of 33 samples, 26 (78.8%) tested positive for M. ovis. The sequencing of positive samples showed 100% identity with multiple M. ovis 16S rDNA sequences. No association was observed between the presence of M. ovis and the FAMACHA© score (p = 0.620). Rhipicephalus (Boophilus) microplus (15/33, 45.4%) was the tick species found on the animals. No significant association between M. ovis infection and presence of ticks (p = 0.4134) and age group (p = 0.4221) was observed. This is the first report of M. ovis infection in sheep from Brazil and only the second report of this pathogen in sheep in Latin America.

Resumo Mycoplasma ovis é um patógeno zoonótico emergente com distribuição mundial e pode causar anemia hemolítica de leve a grave, icterícia e baixo ganho de peso em animais. Embora M. ovis tenha sido descrito em pequenos ruminantes em todo o mundo, os dados sobre M. ovis em ovinos no Brasil são desconhecidos. O objetivo deste estudo foi apresentar o primeiro relato de micoplasmas hemotrópicos em ovinos no Brasil. Avaliamos os fatores associados a essa infecção, como faixa etária, presença de carrapatos e anemia. Amostras de sangue foram coletadas de 33 ovelhas de uma fazenda no sul do Brasil e testadas para hemoplasmas usando a PCR. Das 33 amostras, 26 (78,8%) apresentaram resultado positivo. O sequenciamento das amostras positivas mostrou 100% de identidade com múltiplas sequências de M. ovis 16S rDNA. Não foi observada associação entre a presença de M. ovis e o escore FAMACHA© (p = 0,620). Rhipicephalus (Boophilus) microplus (15/33, 45,4%) foi a espécie de carrapato encontrada nos animais. Não houve associação significativa entre infecção por M. ovis e presença de carrapatos (p = 0,4134) e faixa etária (p = 0,4221). Este é o primeiro relato de infecção por M. ovis em ovinos no Brasil e o segundo relato deste patógeno em ovinos na América Latina.

First molecular detection of Mycoplasma ovis (Hemotropic mycoplasmas) from Sheep in Brazil.

Mycoplasma ovis is an emerging zoonotic pathogen with a worldwide distribution and can cause mild to severe hemolytic anemia, icterus, and poor weight gain in animals. Although M. ovis has been described in small ruminants worldwide, data on M. ovis in sheep in Brazil is unknown. The objective of the present study was to present the first report of hemotropic mycoplasma (HM) in sheep from Brazil. We evaluated factors associated with this infection, such age group, tick presence, and anemia. Blood samples were collected from 33 sheep from a farm in southern Brazil and screened for hemoplasmas using PCR. Out of 33 samples, 26 (78.8%) tested positive for M. ovis. The sequencing of positive samples showed 100% identity with multiple M. ovis 16S rDNA sequences. No association was observed between the presence of M. ovis and the FAMACHA© score (p = 0.620). Rhipicephalus (Boophilus) microplus (15/33, 45.4%) was the tick species found on the animals. No significant association between M. ovis infection and presence of ticks (p = 0.4134) and age group (p = 0.4221) was observed. This is the first report of M. ovis infection in sheep from Brazil and only the second report of this pathogen in sheep in Latin America.

Pupipara (Diptera, Hippoboscidae) in wild birds attended at a rehabilitation center in southern Brazil.

The hippoboscids are cosmopolitan permanent obligate hematophagous ectoparasites of birds, domestic and wild mammals and, occasionally, humans. Some species may act as vectors or hosts of etiological pathogenic agents. The aims of this study were to report on the first cases of Hippoboscidae in Crax blumenbachii and Parabuteo unicinctus; to provide new reports from Brazil on Tyto furcata and Asio stygius parasitized by Icosta americana; to report on individuals of Bubo virginianus, Falco sparverius and Accipiter striatus parasitized by genera Ornithoctona; and to provide new reports on parasitism of O. erythrocephala in the state of Rio Grande do Sul. The birds of prey and C. blumenbachii were attended at a rehabilitation center in Porto Alegre and at a veterinary hospital in Cruz Alta. These new records demonstrate the huge gap that exists regarding studies on avian ectoparasites and highlight potential vectors of hemoparasites for the bird species studied.

Pupipara (Diptera, Hippoboscidae) in wild birds attended at a rehabilitation center in southern Brazil / Pupíparas (Diptera, Hippoboscidae) em Aves Silvestres Atendidas em Centro de Reabilitação no Sul do Brasil

Abstract The hippoboscids are cosmopolitan permanent obligate hematophagous ectoparasites of birds, domestic and wild mammals and, occasionally, humans. Some species may act as vectors or hosts of etiological pathogenic agents. The aims of this study were to report on the first cases of Hippoboscidae in Crax blumenbachii and Parabuteo unicinctus; to provide new reports from Brazil on Tyto furcata and Asio stygius parasitized by Icosta americana; to report on individuals of Bubo virginianus, Falco sparverius and Accipiter striatus parasitized by genera Ornithoctona; and to provide new reports on parasitism of O. erythrocephala in the state of Rio Grande do Sul. The birds of prey and C. blumenbachii were attended at a rehabilitation center in Porto Alegre and at a veterinary hospital in Cruz Alta. These new records demonstrate the huge gap that exists regarding studies on avian ectoparasites and highlight potential vectors of hemoparasites for the bird species studied.

Resumo Os hipoboboscídeos são ectoparasitos hematófagos obrigatórios, permanentes e cosmopolitas de aves, mamíferos domésticos e silvestres e, ocasionalmente, humanos. Algumas espécies podem atuar como vetores ou hospedeiros de agentes patogênicos etiológicos. Os objetivos deste estudo foram relatar os primeiros casos de Hippoboscidae em Crax blumenbachii e Parabuteo unicinctus; fornecer novo relato do Brasil sobre Tyto furcata e Asio stygius parasitados por Icosta americana; relatar indivíduos de Bubo virginianus, Falco sparverius e Accipiter striatus parasitados pelo gênero Ornithoctona; e fornecer novos relatos sobre parasitismo de O. erythrocephala no estado do Rio Grande do Sul. As aves de rapina e C. blumenbachii foram atendidas em um centro de reabilitação em Porto Alegre e em um hospital veterinário em Cruz Alta. Esses novos registros demonstram a enorme lacuna que existe em relação aos estudos sobre ectoparasitas aviários e destacam potenciais vetores de hemoparasitos para as espécies de aves estudadas.

Evaluation of the vector competence of six ixodid tick species for Rangelia vitalii (Apicomplexa, Piroplasmorida), the agent of canine rangeliosis.

Rangelia vitalii is the etiologic agent of canine rangeliosis, a severe piroplasmosis that affects domestic dogs in Brazil, Uruguay and Argentina. While R. vitalii is one of the most pathogenic tick-borne pathogens for dogs in the world, its tick vector has remained unknown. The present study evaluated the vector competence of Rhipicephalus sanguineus sensu lato (both tropical and temperate species), Amblyomma aureolatum, Amblyomma ovale, Amblyomma tigrinum, and Amblyomma sculptum for R. vitalii. These six tick species were selected for the study because they comprise the main tick species infesting dogs within the distribution area of canine rangeliosis in South America. Acquisition feeding of the above six tick species was performed on domestic dogs showing clinical signs of canine rangeliosis, after being experimentally infected through intravenous inoculation or infestation with R. vitalii-infected ticks. Thereafter, engorged ticks were evaluated for transstadial and transovarial passages of R. vitalii through molecular analysis after molting or oviposition and egg hatching. The resultant ticks were evaluated for their competence to transmit R. vitalii to susceptible dogs. Among the six tick species, only A. aureolatum was able to acquire and perpetuate R. vitalii by transstadial and transovarial passages, as demonstrated by >5% infection rates of ticks after hatching or molting. When exposed to transmission feeding, only A. aureolatum ticks were competent to transmit R. vitalii to dogs, which became severely ill, and the results confirmed by molecular methods and blood smear examination to have acquired rangeliosis. Results of the present study, coupled with epidemiological data, indicate that A. aureolatum is a natural vector of R. vitalii. Our results also indicate that R. vitalii is the first Piroplasmorida agent to be transovarially transmitted in Amblyomma ticks.

'Candidatus Mycoplasma haemobos': Transplacental transmission in dairy cows (Bos taurus).

'Candidatus Mycoplasma haemobos' is a haemotropic mycoplasma that can produce various clinical signs in cattle, but abortive potential of the parasite is unknown, as well as the frequency of transplacental transmission in cattle. Thus, the objective of this work was to evaluate the frequency of detection of 'C. M. haemobos' in aborted fetuses and the blood of dairy cows. Blood samples of 22 dairy cows that aborted and pool tissues (brain, lung, heart and liver) of their respective aborted fetuses were tested by conventional PCR. The occurrence of 'C. M. haemobos' DNA in adult animals was 40.9% (9/22) and in the fetuses was 18.2% (4/22). Two fetuses that contained 'C. M. haemobos' DNA were derived from cows which were PCR negative. When stratifying by breed, it was observed that Jersey cows had a higher proportion of positive animals (8/11; 72.7%) as compared to Holstein (1/9; 11.1% P<0.01). The results of this study suggest that this parasite can be transferred via the placenta, but it is not certain if the abortions were due to 'C. M. haemobos'.