First record of Hydatigera taeniaeformis in Geoffroy's cat (Leopardus geoffroyi) in Brazil.

Leopardus geoffroyi (Geoffroy's cat) is a neotropical feline considered globally threatened. In Brazil, it occurs exclusively in the Pampa biome. Its predatory habits contribute to the infection, dispersion, and continuation of the life cycle of various pathogens, including helminths, within ecosystems. However, few studies involving cestodes in wild felines are found in the literature, especially in Brazil. Therefore, we aimed to report the first case of parasitism by Hydatigera taeniaeformis in L. geoffroyi. The helminths were found in the small intestine of the necropsied feline. Specimens were analyzed morphometrically and subjected to molecular analyses for taxonomic identification. The molecular phylogeny based on the analysis of the mitochondrial gene (COX1) allowed the identification of these parasites. Thus, this is the first description of H. taeniaeformis parasitizing L. geoffroyi in Brazil. Consequently, the number of known host species parasitized by this helminth in the country and the world is increased. Additionally, a new molecular sequence is being provided, contributing to the knowledge of Hydatigera in South America.

Molecular identification of Chrysomya albiceps (Diptera: Calliphoridae) in a case of primary cutaneous myiasis in sheep at southern Brazil.

Sheep farming has been growing in Brazil, driven by an expanding consumer market due to greater acceptance of its meat and derivatives. There are several factors that limit sheep production, and one of them is infestation by ectoparasites, which cause stress in animals, weight loss, poor development, low productivity, low quality wool and reduced fertility. Chrysomya albiceps is a species of blowfly belonging to the Calliphoridae family that occurs in neotropical regions, where it causes secondary myiasis. We identified here a rare case of cutaneous myiasis with the presence of tissue lesions caused by C. albiceps in sheep in southern Brazil. We highlight the need to carry out more in-depth studies regarding the biology of these insects, with the aim of proving this atypical behavior for Brazil.

First record of Contracaecum australe (Nematoda: Anisakidae) infecting Neotropic cormorant (Phalacrocorax brasilianus) (Aves: Phalacrocoracidae) in southern Brazil with a phylogenetic analysis.

Phalacrocorax brasilianus (Gmelin, 1789) (Aves: Phalacrocoracidae) is one of the few piscivorous birds inhabiting freshwater and saline environments, being considered one of the most abundant aquatic species in Rio Grande do Sul, Brazil, especially along the state's coastline. It is known that birds are hosts to a wide variety of disease-causing agents, among them, nematodes of the Contracaecum (Anisakidae) have a large number of recognized species. However, little is still known about the occurrence of these parasites in the Southern region of Brazil. Herein we identified for the first time Contracaecum australe Garbin, Mattiucci, Paoletti, González-Acuña, and Nascetti, 2011 (Nematoda: Anisakidae) parasitizing P. brasilianus in Southern Brazil. Nematodes found in the bird's proventriculus were subjected to morphometric analyses, by optical microscopy and scanning electron microscopy, and molecular analyses. Molecular phylogeny based on the analysis of the 18S, ITS-1, 5.8S and ITS-2 genes showed our sequences identical to those of C. australe. Therefore, this is the first record of C. australe in southern Brazil, expanding the geographical distribution of the parasite species in the country. Additionally, new molecular sequences are being provided, contributing to the knowledge of Contracaecum species parasitizing cormorants.

Absence of Leishmania spp. DNA in road-killed wild mammals in Southern Brazil.

Leishmaniasis are neglected diseases transmitted by vectors that affect domestic and wild animals, including humans. Due to its incidence and lethality, this zoonosis is a worrying public health problem, making it essential to identify all links in the transmission chain. Infection of wild mammals by Leishmania spp. remains poorly understood, especially in southern Brazil. Therefore, the objective was to research, using the PCR technique, the presence of Leishmania spp. DNA in road-killed wild mammals in Southern Brazil. Carcasses of 96 animals were collected from highways in the Pelotas microregion, Rio Grande do Sul, southern Brazil and subjected to necropsies. Tissue fragments (spleen, skin, liver, kidney, heart, lung, lymph nodes, bone marrow and blood) were collected and genomic DNA was extracted. PCR protocols targeting the ITS1, kDNA and 18S genes were tested. We found no evidence of Leishmania spp. circulation in the studied population. However, epidemiological studies like this one are of great relevance, as they allow monitoring of the occurrence of pathogens and help identify possible risk areas. As these animals act as epidemiological markers for the presence of the microorganism, studies must be carried out continuously to understand whether there are sources of infection in the region.

Control of Rhipicephalus microplus tick larvae in the field based on distancing from the host - Lone tick system.

The aim of this study was to evaluate a host-tick distancing control system using Rhipicephalus microplus in cattle. Its impact on the profiles of the pathogens Anaplasma marginale and Babesia bigemina was also assessed using enzyme-linked immunosorbent and real-time polymerase chain reaction. Experiments were conducted in Mato Grosso do Sul, Brazil. A total of 37 Senepol animals with natural tick infestation were used in this study. The ticks on the animals were counted, and pasture rotation was performed at 28-day intervals. This approach was repeated until the end of the experiment to maintain a low number of ticks, resulting in a final average of 6.3 ticks/host. Cattle maintained an immune response to the cattle tick fever (CTF) agents. DNA copies of A. marginale (X̄ 1.1) and B. bigemina (X̄ 0.05) did not vary significantly with the decrease in tick count throughout the study period. Based on these results, we conclude that a distancing period of 84 days between ticks and cattle in conditions of the Cerrado biome can control ticks in animals and maintain enzootic stability. Thus, it is possible to create productive breeds with lower tick control costs.

First reported case of Trichomonas gallinae infection in red-breasted toucan (Ramphastos dicolorus).

This study reports the infection and diagnosis of the protozoan morphologic complex Trichomonas gallinae in a baby red-breasted toucan (Ramphastos dicolorus). Nodular lesions on the soft palate and edema in the oral cavity were observed macroscopically. Microscopically, a granuloma with multiple layers of necrosis interspersed with inflammatory polymorphonuclear infiltrates was observed. Parasitism was confirmed by parasitological diagnosis, isolation of the flagellates in culture medium, and Polymerase Chain Reaction (PCR) using 5.8S ribosomal RNA (rRNA). Flanking internal transcribed spacer (ITS) gene regions were amplified by polymerase chain reaction, and the sequences were analyzed phylogenetically using MEGA 11 software. Phylogenetic analysis based on ITS1/5.8S rRNA/ITS2 sequences demonstrated high nucleotide identity with two Trichomonas sequences available in GenBank, which were more closely related to T. vaginalis (99%) than to T. gallinae (98%). In addition to being potential transmitters of this protozoan, rigorous monitoring of infectious and parasitic diseases in wild bird populations is essential for their preservation. The forms of transmission of Trichomonas sp. favor the occurrence of the disease in many non-Columbiformes species, which is essential for the monitoring of this disease in wild birds.

Absence of Leishmania spp. DNA in road-killed wild mammals in Southern Brazil / Ausência de DNA de Leishmania spp. em mamíferos silvestres atropelados no Sul do Brasil

Abstract Leishmaniasis are neglected diseases transmitted by vectors that affect domestic and wild animals, including humans. Due to its incidence and lethality, this zoonosis is a worrying public health problem, making it essential to identify all links in the transmission chain. Infection of wild mammals by Leishmania spp. remains poorly understood, especially in southern Brazil. Therefore, the objective was to research, using the PCR technique, the presence of Leishmania spp. DNA in road-killed wild mammals in Southern Brazil. Carcasses of 96 animals were collected from highways in the Pelotas microregion, Rio Grande do Sul, southern Brazil and subjected to necropsies. Tissue fragments (spleen, skin, liver, kidney, heart, lung, lymph nodes, bone marrow and blood) were collected and genomic DNA was extracted. PCR protocols targeting the ITS1, kDNA and 18S genes were tested. We found no evidence of Leishmania spp. circulation in the studied population. However, epidemiological studies like this one are of great relevance, as they allow monitoring of the occurrence of pathogens and help identify possible risk areas. As these animals act as epidemiological markers for the presence of the microorganism, studies must be carried out continuously to understand whether there are sources of infection in the region.

Resumo As leishmanioses são doenças negligenciadas, transmitidas por vetores que acometem animais domésticos e silvestres, incluindo os humanos. Devido a sua incidência e letalidade, essa zoonose consiste em um problema de saúde pública preocupante, sendo fundamental a identificação de todos os elos da cadeia de transmissão. A infecção de mamíferos silvestres por Leishmania spp. permanece pouco compreendida, especialmente no sul do Brasil. Portanto, objetivou-se pesquisar, por meio da técnica de PCR, a presença de DNA de Leishmania spp. em mamíferos silvestres atropelados no Sul do Brasil. Carcaças de 96 animais foram coletadas, em rodovias da microrregião de Pelotas, Rio Grande do Sul, sul do Brasil e submetidas a necropsias. Fragmentos de tecidos (baço, pele, fígado, rim, coração, pulmão, linfonodos, medula óssea e sangue) foram coletados e o DNA genômico foi extraído. Protocolos de PCR visando os genes ITS1, kDNA e 18S foram testados. Não foram encontradas evidências de circulação de Leishmania spp. na população estudada. Porém, estudos epidemiológicos como este são de grande relevância, pois permitem monitorar a ocorrência de patógenos e auxiliam na identificação de possíveis áreas de risco. Como esses animais atuam como marcadores epidemiológicos da presença do microrganismo, estudos devem ser realizados continuamente, para entender se existem fontes de infecção na região.

Association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of taurine heifers.

The aim of this study was to investigate the association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of pregnant and non-pregnant taurine heifers. Blood samples from 94 females were collected on the first day (D-10) of timed artificial insemination (TAI) protocol and on pregnancy diagnosis (D+34). Hematological parameters were determined and compared between pregnant (PG) and non-pregnant (NPG) heifers, and within group at different sampling days. Real-time PCR (qPCR) was used to determine A. marginale and Babesia bovis infection, and for absolute quantification of Babesia spp. between PG and NPG groups. Correlation analysis was performed between the number of gDNA copies (CN) of Babesia spp. and hematological parameters. On D-10, mean hemoglobin concentration was higher for NPG, and hematocrit and total plasma protein were higher on D+34 for both groups. There was no difference in Babesia spp. CN between groups. In the first qPCR, all heifers were positive for A. marginale and B. bovis. Significant correlations were found between hemoglobin and erythrocyte and between hemoglobin and hematocrit (r = 0.8082 and r = 0.3009, respectively). Low levels of A. marginale and Babesia spp. did not affect hematological parameters of chronically infected pregnant and non-pregnant taurine heifers.

Seasonal fluctuations of Babesia bigemina and Rhipicephalus microplus in Brangus and Nellore cattle reared in the Cerrado biome, Brazil.

BACKGROUND: The tick Rhipicephalus microplus (Ixodida: Ixodidae, Canestrini, 1888) causes substantial economic and health losses in cattle production and is the main vector of Babesia bigemina (Piroplasmidae: Babesidae, Smith & Kilborne, 1893). Babesia bigemina is responsible for a tick-borne disease known as babesiosis that can cause hemolytic anemia, fever and death. In the study reported here, we investigated the relationship between the number of ticks per animal and the number of B. bigemina cytochrome b gene (cbisg) copies in the blood of Brangus and Nellore cattle reared without acaricidal treatment in the Brazilian Cerrado biome over a 1-year period. METHODS: Ticks on 19 animals (9 Brangus and 10 Nellore cattle) were counted every 18 days, and blood was collected every 36 days for 12 months. Serological samples were analyzed with an indirect enzyme-linked immunosorbent assay, and genomic DNA was analyzed by conventional PCR and quantitative PCR. The PCR products were sequenced by the Sanger method. RESULTS: The Brangus and Nellore breeds showed similar weight development and no clinical signs of babesiosis. Statistically significant differences (P < 0.05) between the breeds were observed for the number of ticks and the number of B. bigemina cbisg gene copies. CONCLUSIONS: No correlation between the number of ticks and the number of circulating copies of cbisg was observed, although Nellore cattle presented with fewer ticks than Brangus cattle and the number of cbisg copies was higher for Nellore cattle than for Brangus cattle.

Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil.

Neospora caninum is an obligate intracellular protozoan with canids (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) as its definitive hosts. The objective of this study was to detect anti-N. caninum antibodies in pregnant women seen at referral center for prenatal screening in the state of state Mato Grosso do Sul, Brazil. A total of 188 serum samples from pregnant women provided by the Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE) were subjected to IFA test and western blot analysis. The samples were divided into three groups: 23/99 samples from the seropositive group for toxoplasmosis were positive for anti-N. caninum IgG antibodies, and 9/99 positive for IgM; in the HIV group, 7/33 were positive for IgG; and in the HIV+toxoplasmosis group, 13/56 were positive for IgG and two positive for IgM. The seropositivity for IgG was assessed by western blot by testing 43 IFA test positive samples using rNcSRS2 (Nc-p43) as antigen. The serological results of the present study suggest that exposure of these pregnant women to the parasite N. caninum and presence of IgM antibodies are indicative of recent infection. Further studies are needed to establish the possibility of active infection.

Correlation between Rhipicephalus microplus ticks and Anaplasma marginale infection in various cattle breeds in Brazil.

The tick Rhipicephalus microplus is responsible for the transmission of Anaplasma marginale, which causes hemolytic anemia, abortion, decreased production, and mortality in cattle in Brazil. However, A. marginale can also persist in cattle herds without any clinical signs. This study investigated the relationship between the number of ticks present on each cattle and the circulating number of A. marginale msp1ß gene copies in the blood of Brangus and Nellore cattle reared in the Brazilian Cerrado through a year period. Twenty-three animals (11 Brangus and 12 Nellore) were raised for 12 months with ticks counted every 18 days, and blood collected every 36 days. Blood sera was used for total antigen iELISA, genomic DNA was extracted from whole blood by the phenol/chloroform method and then analyzed by PCR to confirm A. marginale presence with the msp5 gene. Positive samples were quantified by qPCR using msp1ß gene. Brangus cattle presented 4.5 fold more ticks than Nellore group. Although Brangus cattle carried a higher overall A. marginale msp1ß gene presence than Nellore cattle, no relationship of tick count and copy number could be achieved due to high variability in copy number. Moreover, both breeds showed similar weight gain and a similar serological pattern throughout the year. None of the animals showed any clinical signs of anaplasmosis during the experimental period, indicating that a low level of tick infestation may be sufficient to maintain a stable enzootic situation.

Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil / Avaliação sorológica de Neospora caninum em gestantes atendidas em centro de referência para triagem pré-natal em Mato Grosso do Sul, Brasil

Abstract Neospora caninum is an obligate intracellular protozoan with canids (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) as its definitive hosts. The objective of this study was to detect anti-N. caninum antibodies in pregnant women seen at referral center for prenatal screening in the state of state Mato Grosso do Sul, Brazil. A total of 188 serum samples from pregnant women provided by the Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE) were subjected to IFA test and western blot analysis. The samples were divided into three groups: 23/99 samples from the seropositive group for toxoplasmosis were positive for anti-N. caninum IgG antibodies, and 9/99 positive for IgM; in the HIV group, 7/33 were positive for IgG; and in the HIV+toxoplasmosis group, 13/56 were positive for IgG and two positive for IgM. The seropositivity for IgG was assessed by western blot by testing 43 IFA test positive samples using rNcSRS2 (Nc-p43) as antigen. The serological results of the present study suggest that exposure of these pregnant women to the parasite N. caninum and presence of IgM antibodies are indicative of recent infection. Further studies are needed to establish the possibility of active infection.

Resumo Neospora caninum é um protozoário intracelular obrigatório que possui os canídeos (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) como seus hospedeiros definitivos. O objetivo deste trabalho foi detectar anticorpos anti-N. caninum em gestantes, atendidas em centro de referência para triagem pré-natal, em Mato Grosso do Sul, Brasil. Um total de 188 amostras de soro de gestantes, cedidas pelo Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE), foram submetidas ao teste de RIFI e western blot para a detecção de anticorpos anti-N. caninum. As amostras foram divididas em três grupos: 23/99 amostras do grupo soropositivo para toxoplasmose demonstraram positividade para anticorpos IgG anti-N. caninum e 9/99 positivos para IgM. No grupo HIV 7/33 apresentaram positividade para IgG. No grupo HIV+Toxoplasmose 13/56 apresentaram positividade para IgG e duas para IgM. A soropositividade para IgG foi avaliada por western blot, testando 43 amostras positivas para RIFI, usando-se rNcSRS2 (Nc-p43) como antígeno. Os resultados sorológicos do presente estudo sugerem exposição dessas gestantes ao parasita N. caninum e a positividade para anticorpos IgM são indicativos de infecção recente. Mais estudos na área são necessários para estabelecer a possibilidade de infecção ativa.