Serological evidence of Ehrlichia minasensis infection in Brazilian dogs.

Ehrlichia spp. are important tick-borne pathogens of animals in Brazil, and Ehrlichia canis is the most prevalent species infecting dogs. Moreover, Ehrlichia minasensis has also recently been identified as a novel ehrlichial agent that infects cattle in Brazil. The objective of this study was to determine whether dogs could be infected by E. minasensis. To investigate this possibility, sera (n = 429) collected from dogs in the Pantanal region were retrospectively analyzed for the presence of antibodies against E. canis and E. minasensis. Canine sera were screened by two isolates of E. canis in indirect immunofluorescence assay (IFA) and the majority (n = 298; 69.4%) had antibodies with endpoint titers ranging from 80 to 327,680. In order to further confirm E. canis-specific antibodies, IFA positive sera were analyzed by ELISA using E. canis-specific peptides (i.e. TRP19 and TRP36 US/BR/CR), which detected E. canis antibodies in 80.2% (239/298) of the dog sera. Fifty-nine (13.7%) samples had detectable antibodies to E. canis by IFA but were negative by E. canis peptide ELISA. These sera were then tested by E. minasensis IFA (Cuiaba strain) as antigen and 67.8% (40/59) were positive (titers ranging from 80 to 20,480). Eleven sera had antibody titers against E. minasensis at least two-fold higher than observed for E. canis and suggests that these dogs were previously infected with E. minasensis. The results of the present study suggest that multiple ehrlichial agents infect dogs in Brazil, which highlights the need to consider different Ehrlichia spp. in Brazilian dogs, particularly in areas where dogs are frequently exposed to multiple tick species. This investigation is the first to provide serologic evidence of E. minasensis infection in dogs from Brazil.

Detection of Ehrlichia sp. in Amblyomma sculptum parasitizing horses from Brazilian Pantanal wetland.

Bacteria of the genus Ehrlichia are transmitted by ticks and also are an important cause of infection in wild and domestic mammals. Infection with Ehrlichia spp. has been reported in horses, especially in the USA, Nicaragua and Brazil. In this study, we report the parasitism by Amblyomma sculptum, Rhipicephalus microplus and Dermacentor nitens ticks in horses from a ranch located in south Pantanal wetland. Molecular and serological analyzes to determine infection by Ehrlichia spp. in horses and their respective ticks were carried out. A total of 12 horses were submitted to blood collection to investigate antibodies by indirect immunofluorescence assay (IFA) using Ehrlichia canis crude antigens and to be tested by polymerase chain reaction (PCR) in order to amplify fragments of the 16S rRNA, dsb, groEL and sodB gene of Ehrlichia spp. A total of 164 tick specimens were removed from horses, stored in isopropanol and later identified as D. nitens, A. sculptum and R. microplus. DNA from ticks were extracted and subjected to the same PCR assays to detect Ehrlichia spp. Anti-Ehrlichia spp. antibodies were detected in five/12 (41.7 %) horses by IFA, with antibody titers ranging from 40 to 160. All horse DNA samples were negative for the 16S rRNA, dsb, groEL and sodB of Ehrlichia spp. One A. sculptum female was positive to all target genes of Ehrlichia. This tick was parasitizing an Ehrlichia-seropositive horse with antibody titer of 80. Nucleotide sequences of 16S rRNA, dsb, groEL and sodB genes showed close relationship with different strains of Ehrlichia detected in wild mammals, Amblyomma ticks and horses from Brazil and Argentina. Detection of anti-Ehrlichia sp. antibodies suggests that horses have been exposed to an ehrlichial agent in the Pantanal. Future studies on Ehrlichia infection should be carried out to better elucidate and to bring new information about equine ehrlichiosis, since these animals are important hosts of ticks in the Brazilian Pantanal wetlands.

High genetic diversity of paramyxoviruses infecting domestic cats in Western Brazil.

Feline morbillivirus was discovered in 2012 in cats from Hong Kong, and it was initially found to be associated with chronic kidney disease. Although subsequent molecular surveys showed a common occurrence in cat populations from distinct countries, there were controversial results regarding the relationship between viral shedding through urine and reduced kidney function. In this study, 276 domestic cats of diverse origins from Western Brazil had their urine evaluated for the presence of paramyxoviral RNA by reverse transcription seminested PCR and direct sequencing. Additionally, a selected Brazilian feline morbillivirus strain was isolated in Crandell Rees feline kidney cells, and a nearly complete genome sequence was obtained. To assess the kidney function of all cats, serum biochemistry screening and standard urinalysis were performed. Our results revealed a relatively high paramyxovirus-positive rate (34.7%) in the evaluated cats although there was not a statistical association between the shedding of viral RNA through urine and kidney disease. Direct sequencing of partial fragments of the L gene demonstrated high genetic diversity among strains detected in cats in this study, since both feline morbillivirus RNA and feline paramyxovirus RNA were frequently shed in urine. Phylogenetic reconstruction based on partial amino acid sequences of the L gene showed that Brazilian feline paramyxovirus strains were genetically diverse since they grouped into two distinct subclusters; one subcluster contained three strains identified in Germany, while the second contained Japanese strain 163, which was recently classified in the Jeilongvirus genus of the Paramyxoviridae family. In contrast, the Brazilian feline morbillivirus strain FeMV/BR_Boni, herein characterized by nearly complete genome sequencing, was classified in the Morbillivirus genus with other strains previously identified as genotype 1. In conclusion, urinary excretion of diverse paramyxoviral RNA is frequent in cats of different origins from Western Brazil, but viral infection is not related to altered kidney function.

Uncommon Ehrlichia canis infection associated with morulae in neutrophils from naturally infected dogs in Brazil.

Ehrlichia and Anaplasma species are the most common tick-borne disease (TBD) pathogens in dogs worldwide. Ehrlichia canis, the aetiological agent of the Canine Monocytic Ehrlichiosis (CME), is known to replicate within the cytoplasm of mononuclear cells into clusters of organisms called morulae. However, detection of morulae in neutrophils is commonly observed in dogs infected by Ehrlichia ewingii or Anaplasma phagocytophilum. We report uncommon clinical cases of canine ehrlichiosis presenting morulae compatible with E. ewingii and A. phagocytophilum in dogs from two distinct regions of Brazil. Eight dogs were admitted to two veterinary teaching hospitals from Brazil, showing clinical or haematological signs suggestive of TBD. Blood or peritoneal fluid was withdrawn for haematological and cytologic analysis. All samples were evaluated by PCR assays for Ehrlichia and Anaplasma using genus-specific primers for dsb, 16S rRNA and groEL genes, followed by sequencing. Samples were also evaluated by nested PCR assays for the 16S rRNA gene of E. ewingii and groEL gene of A. phagocytophilum and Anaplasma platys. Seven dogs revealed thrombocytopenia, six dogs had monocytosis and five presented lymphopenia and anaemia. All dogs showed morulae structures compatible with Ehrlichia spp. in neutrophils and were PCR-positive for the dsb and 16S rRNA gene fragments of Ehrlichia, with sequences showing 100% identity with multiple E. canis sequences deposited in the GenBank™. Sequencing of 16S rRNA and groEL gene fragments from one PCR-positive dog showed 100% identity with A. platys. Overall, our data suggest that in endemic regions for E. canis, that is Brazil, the presence of morulae in neutrophils may indicate infection by this bacterium. Herein, morulae were also found in neutrophils present in the peritoneal fluid of a dog. Also, this is the first report of E. canis and Hepatozoon canis co-infection in neutrophils from naturally infected dogs confirmed by DNA sequencing.

Immunohistochemical characterization of cutaneous leishmaniasis in cats from Central-west Brazil.

Feline leishmaniasis (FeL) is an emerging infectious disease of cats caused by Leishmania infantum with global distribution. This study investigated the cause of chronic progressive cutaneous lesions in two cats from Central-west Brazil by using cytological, histopathologic, and immunohistochemical (IHC) analyses. Clinically, both cats had ulcerative cutaneous lesions at the nasal planum and ear resulting in a tentative diagnosis of squamous cell carcinoma (SCC). Moreover, both cats had varying degrees of onychogryphosis. However, cytology revealed chronic inflammatory reactions associated with intralesional amastigotes; histopathology confirmed chronic ulcerative dermatitis associated with intralesional and intracytoplasmic parasitic organisms consistent with amastigotes of Leishmania spp. within histiocytes. The IHC assay demonstrated that the intralesional parasitic structures identified by cytology and histopathology were immunoreactive to antigens of Leishmania spp., confirming the participation of this infectious disease agent in the development of the cutaneous lesions of these cats. The observation of onychogryphosis must be highlighted, since this lesion is frequently observed in dogs with visceral leishmaniasis but is underreported in FeL. Collectively, the pathologic and IHC findings of the chronic cutaneous disease confirmed active infections due to Leishmania spp. in these cats. Additionally, FeL with associated lesions to the ear and nasal planum must be considered as differential diagnosis for SCC in cats.

First report of feline morbillivirus in South America.

Feline morbillivirus was first identified in healthy and diseased stray cats captured in Hong Kong. Recently, it was demonstrated that the virus circulates within cat populations in Japan, Italy, Germany, and the USA. Importantly, an association between feline morbillivirus infection and chronic kidney disease was suggested by histological analysis of kidney tissue of infected cats. The aim of this study was to verify the presence and examine the genetic diversity of feline morbilliviruses associated with infections of domestic cats in Brazil. Seventeen cats without clinical manifestations of urinary tract diseases from a multi-cat household and 35 random client-owned cats admitted to the Teaching Veterinary Hospital for a variety of reasons were evaluated for paramyxoviral infection and the presence of uropathy. A fragment of the paramyxoviral L gene was amplified from urine samples using a reverse transcription semi-nested PCR assay. For the first time, we detected a feline morbillivirus strain that was genetically related to viral strains previously characterized in Japan in urine samples from cats in South America, in Brazil. This together with the recent description of feline morbillivirus identification within cat populations in the USA, suggests a possible widespread distribution of this viral agent on the American continent. Our data demonstrated feline morbillivirus RNA shedding mostly in the urine of cats without clinical, laboratorial, or ultrasonographic signs of urinary tract diseases. In contrast to previously published findings that associated feline morbillivirus infection with chronic kidney disease, we did not observe a clear relationship between feline morbillivirus RNA shedding in urine and kidney disease in the cats evaluated.

Evitando erros de diagnóstico em pacientes com paraparesia/plegia aguda com ênfase no botulismo / Avoiding misdiagnosis in patients with acute paresis/paralysis with emphasis on botulism

[...] Mortes envolvendo botulismo geralmente estão relacionadas com a falha em reconhecer a doença e a gravidade, refletindo no retardo em iniciar a terapia, seja esta suporte ou específica (BARSANTI, apud GREENE, 2006). A diferenciação dos casos é extremamente importante, assim o objetivo geral foi o de criar um sistema padrão para abordagem clínica primária em casos de manifestações neuromusculares agudas, evitando erros de diagnóstico nesse tipo de emergências neurológicas, focado no diagnóstico de botulismo.

Seroprevalence of Toxoplasma gondii infection in swine matrices in Nova Mutum and Diamantino, Mato Grosso, Brazil.

This report aimed to assess the seroprevalence of Toxoplasma gondii infection in 708 swine matrices in Nova Mutum and Diamantino in the state of Mato Grosso, Central-West Brazil. Serum samples were examined by indirect fluorescent antibody test (IFAT). It was found a seroprevalence of 12.8%, considering titers ≥ 64. Therefore, the data reinforce the need for appropriate management of swine raising to minimize the risk of infection of pigs with T. gondii.

Seroprevalence of Toxoplasma gondii infection in swine matrices in Nova Mutum and Diamantino, Mato Grosso, Brazil / Soroprevalência de infecção por Toxoplasma gondii em matrizes suínas em Nova Mutum e Diamantino, Mato Grosso, Brasil

This report aimed to assess the seroprevalence of Toxoplasma gondii infection in 708 swine matrices in Nova Mutum and Diamantino in the state of Mato Grosso, Central-West Brazil. Serum samples were examined by indirect fluorescent antibody test (IFAT). It was found a seroprevalence of 12.8 percent, considering titers >64. Therefore, the data reinforce the need for appropriate management of swine raising to minimize the risk of infection of pigs with T. gondii.

No presente trabalho, objetivou-se avaliar a soroprevalência da infecção por Toxoplasma gondii, em 708 matrizes suínas dos municípios de Nova Mutum e Diamantino do Estado de Mato Grosso, Brasil. As amostras de soro foram examinadas por meio da reação de imunofluorescência indireta (RIFI). Foi encontrada a frequência de 12,8 por cento de soros positivos, com diluições iguais ou superiores a 64. Portanto, os dados obtidos reforçam a necessidade de um manejo de criação adequado, visando à minimização do risco de infecção de suínos por T. gondii.