Case report of hepatic calodiosis in a rescued dog.

Calodium hepaticum (syn. Capillaria hepatica) is a nematode responsible for parasitic zoonosis that can cause granulomatous lesions in the liver. Although murids are the main hosts, C. hepaticum has been documented in a wide range of mammals. Here, we report a case of hepatic calodiosis in a 6-month-old dog that was rescued in the municipality of Cuiabá, Mato Grosso state, midwestern Brazil. Even after being attended at a Veterinary Hospital, the case resulted in animal death. Necroscopic examination revealed significant macroscopic changes in the spleen, liver, and encephalus. Microscopic examination revealed parasitic eggs, identified as Calodium cf. hepaticum, that were rounded to oval with a bilamellar cuticle, radially striated, and with bipolar plugs. Considering the difficulty to diagnose this parasite, and the existence of diverse possible hosts in Mato Grosso due to the biodiversity of the state, further investigation is required to understand the true prevalence of C. hepaticum and the importance of this nematode in public health.

Molecular detection and genetic characterization of Ehrlichia canis and Ehrlichia sp. in neotropical primates from Brazil.

The Anaplasmataceae family includes obligate, arthropod-transmitted intracellular bacteria that can be zoonotic and potentially fatal. Studies focusing on the interaction between neotropical primates and the agents of this family are scarce. The present study aimed to identify agents of the Anaplasmataceae family in the whole blood of free-living and captive neotropical primates in the State of Mato Grosso, Central-West Brazil. Thirty-eight samples of six nonhuman primate (NHP) species were collected in seven municipalities and analysed through polymerase chain reaction (PCR), nucleotide sequencing, and phylogenetic analysis of the dsb, groEL, 16S rRNA, and gltA genes. DNA fragments similar to those of Ehrlichia canis were detected in Sapajus apella and Ehrlichia chaffeensis from Mico melanurus. The sequences generated in this study and homologous sequences retrieved from GenBank® were used for phylogenetic analyses to characterize the Ehrlichial agents detected in NHPs. The agents were then grouped into clades corresponding to different isolates from the NHP species. In addition, an Anaplasma sp. closely related to Anaplasma marginale was identified in two S. apella individuals. These findings shed light on the susceptibility of neotropical NHPs to Anaplasmataceae agents. These bacteria are known to be transmitted by ticks, which can also serve as possible sources of infection for other animals, including humans.

MORPHOLOGICAL AND GENETIC CHARACTERIZATION OF DIDELPHONEMA LONGISPICULATA (NEMATODA: SPIRUROIDEA) IN THE BLACK-EARED OPOSSUM DIDELPHIS MARSUPIALIS.

Didelphonema longispiculata (Hill, 1939), a gastric nematode parasite of the black-eared opossum, Didelphis marsupialis Linnaeus, 1758, collected from 2 municipalities of Mato Grosso state, Brazil, in the ecotone region of the Amazon and Cerrado biomes was analyzed with integrative taxonomy using light and scanning electron microscopy (SEM) for morphological studies and sequencing of the 18S small subunit ribosomal RNA for phylogenetic inference through maximum likelihood and Bayesian phylogenetic inference. Here details of the helminth surface, oral aperture with octagonal border, pseudo- and inter-labia, amphids, external cephalic papillae, 2 dorsal and ventral internal plates distally indented, and stoma with strongly chitinized wall are presented. Caudal male papillae, spicules, female vulva, anus, and caudal tip were detailed using SEM. Morphological characteristics and phylogenetic data corroborated the taxonomic placement of the genus Didelphonema within the subfamily Ascaropsinae.

High prevalence of anti-Toxoplasma gondii antibodies in beef cattle in Midwestern Brazil.

The present study aimed to determine the seroprevalence of anti-Toxoplasma gondii antibodies using an indirect immunofluorescence assay (IFA-IgG) in 3,814 cows aged ≥ 24 months belonging to 353 beef farms in the Mato Grosso state. Additionally, we aimed to identify the possible risk factors associated with seropositivity in Mato Grosso, which encompasses three biomes (Amazon, Cerrado, and Pantanal) of Brazil. Analysis of 3,814 samples observed that 1,307 animals were positive for anti-T. gondii antibodies (IFA-IgG ≥ 64), with an animal-level seroprevalence of 34.27%. Herd-level seroprevalence was 92.07%. In the animal-level model, cows raised in the Amazon and Pantanal biomes and breed studies on European and Zebu or hybrid were found to be at risk for T. gondii seropositivity. Prevalence of anti-T. gondii antibodies in cows destined for human consumption was found to be widely distributed throughout the entire study area. Further studies are required to assess the impact of beef in the possible transmission of toxoplasmosis to humans residing in the region and establish additional control measures for this protozoan mainly on beef herds raised in the Amazon and Pantanal biomes, where the highest values of seroprevalence were observed.

Frequency of Giardia duodenalis infection and its genetic variability in dogs in Cuiabá, Midwest Brazil.

INTRODUCTION: Giardia duodenalis, a unicellular, eukaryotic, and flagellated protozoan, presents two evolutionary forms in its life cycle, namely, trophozoites and cysts. It causes diarrhea in humans, dogs, cats, rodents, and ungulates. Despite being morphologically similar, the isolates of G. duodenalis are genetically diverse, affecting the stability and unanimity of taxonomic classification. Since different Giardia assemblages may occur within one isolate, multilocus genotyping is recommended for the genetic identification. METHODOLOGY: To determine the frequency of G. duodenalis infections in domiciled dogs in Cuiabá Municipality (State of Mato Grosso, Midwestern Brazil) and characterize its genetic variability, fecal samples were collected from 147 dogs. RESULTS: Overall, 6.8% (10/147) of the samples presented cysts of G. duodenalis, which sequencing and genotypic characterization using tpi and gluD revealed assemblages C and A, genetic grouping of G. duodenalis. Only three samples amplified by tpi and one sample amplified by gluD. CONCLUSIONS: The risk factors age, gender, breed, diet and the presence of other dogs in the same house were not correlationated with giardiasis. The host-specific and zoonotic genotype warns of the risk of inter and intraspecies transmission and it provides, for the first time, information about genetic characterization of G. duodenalis isolates in dogs in Cuiabá, Midwest region of Brazil.

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazils western Amazon region

Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

Resumo Trypanosoma (Megatrypanum) theileri é um protozoário flagelado que infecta ruminantes e apresenta alta diversidade genética. Neste estudo, investigamos as taxas de prevalência deste protozoário com base na hemocultura e no diagnóstico molecular. Os isolados de T . theileri obtidos foram caracterizados pelos marcadores moleculares SSU rDNA e gGAPDH e o diagnóstico molecular foi baseado no gene do tipo Catepsina L (PCR-TthCATL). O PCR-TthCATL e a hemocultura indicaram uma taxa de prevalência total de 8,13% e a sequência derivada do gene Catepsina L denominada IB de T. theileri foi identificada pela primeira vez em bovinos da Amazônia Ocidental, bem como a IF no Brasil. Também descrevemos uma possível nova sequência derivada da PCR-TthCATL em bovinos, designada IL.

Molecular survey and genetic diversity of piroplasmids in equids from Midwestern Brazil

Abstract We evaluated the distribution of piroplasmids in equids from the Mato Grosso state in Midwestern Brazil using molecular methods and the interspecific genetic diversity. For this, 1,624 blood samples of equids from 973 farms were examined by PCR, using primer pairs that amplify a fragment of the genes rap-1 and ema-1 of Babesia caballi and Theileria equi, respectively. For molecular characterization and phylogenetic studies, 13 and 60 sequences of the rap-1 and ema-1 genes, respectively, were used to build a dendogram using maximum parsimony. B. caballi and T. equi were detected in 4.11% and 28.16% of the farms, respectively, and molecular prevalence was 2.74% for B. caballi and 25.91% for T. equi. The location of the farms and animals raised in the Pantanal ecoregion influence the probability of equids testing positive for B. caballi and T. equi . Moreover, age and herd purpose were variables significantly associated with T . equi infection. The sequences of B. caballi presented 1.95% intraspecific variability, contrasting with 2.99% in T. equi. Dendrograms for both species demonstrated the presence of subgroups with high values of support of branches. However, it is not possible to associate these groups with geographic origin and/or ecoregion.

Resumo Foi avaliada a distribuição de piroplasmídeos em equídeos do Estado de Mato Grosso, no Centro-Oeste do Brasil, utilizando-se métodos moleculares e a diversidade genética interespecífica. Para isso, 1.624 amostras de sangue de equídeos de 973 fazendas foram examinadas pela PCR, usando pares de oligonucleotídeos que amplificam um fragmento dos genes rap-1and ema-1 de Babesia caballi e Theileria equi, respectivamente. Para caracterização molecular e estudos filogenéticos, foram utilizadas 13 e 60 sequências dos genes rap-1 e ema-1, respectivamente, para construção de um dendograma utilizando máxima parcimônia. B. caballi e T . equi foram detectados em 4,11% e 28,16% das fazendas, respectivamente, e a prevalência molecular foi de 2,74% para B. caballi e 25,91% para T. equi. A localização das fazendas e animais criados na ecorregião do Pantanal influenciam a probabilidade de equídeos serem positivos para B. caballi e T. equi. Além disso, idade e propósito do rebanho foram variáveis, significativamente, associadas à infecção por T. equi. As sequências de B . caballi apresentaram variabilidade intraespecífica de 1,95%, contrastando com 2,99% em T. equi. Dendrogramas para ambas as espécies demonstraram a presença de subgrupos com altos valores de sustentação dos ramos. No entanto, não é possível associar esses grupos com origem geográfica e/ou ecorregião.

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazils northern Pantanal wetland

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

Molecular detection of tick-borne protozoan parasites in a population of domestic cats in midwestern Brazil.

Some tick-borne pathogens that infect domestic cats have been considered emergent in veterinary medicine. Occurrences of Hepatozoon spp., Babesia spp. and Cytauxzoon spp. have been described in several regions of Brazil. This paper offers a comprehensive analysis of the 18S rRNA gene of a Hepatozoon sp. strain detected in domestic cats in the metropolitan area of Cuiabá, in Midwestern Brazil. Based on a molecular analysis, we detected the presence of Hepatozoon species circulating among cats in this region. The aforementioned strain is closely related to other isolates of H. felis detected in wild felids. Moreover, a phylogenetic analysis indicates that this genotype is grouped into a clade of 18S rRNA sequences previously described for the genus Hepatozoon in wild felids around the world. Hepatozoon felis strains detected in cats from Spain and Israel showed, respectively, 98% and 97% identity to our sequence and are clustered on a separate branch of the phylogenetic tree. This finding suggests a high diversity of Hepatozoon genotypes occurring in cats in Europe and South America. None of the analyzed cats were positive for Babesia spp. or Cytauxzoon spp. by PCR analysis.