Serum proteins and electrophoretic profile in horses undergoing crotalid venom hyperimmunization.

Indisputably, the use of antivenoms for the treatment of snakebite envenoming is beneficial for the victims. However, there are few studies addressing the effect of long-term hyperimmunization in inoculated horses. It is known that the injection of snake venoms and adjuvants leads to local and systemic reactions in horses, but little is known about the response of inflammatory proteins. The aim of this study was to evaluate serum proteins and the electrophoretic profile of horses undergoing crotalid venom hyperimmunization. Twenty horses were divided into two groups: an inoculated group, comprising ten horses that were already being used for production of a Crotalus sp. antivenom, and a control group, comprising ten animals that had never been used for hyperimmunization. All animals were clinically healthy and without laboratory abnormalities. Total protein and albumin concentrations were measured in serum. Serum globulins were obtained by calculation. Plasma fibrinogen estimates were determined by the heat precipitation method. Serum proteinograms were obtained using agarose gel electrophoresis. The results revealed a significant increase in the concentrations of total serum proteins, globulins, and ß-globulins in the inoculated group, exceeding the reference values. There were slight increases in the α-1- and α-2-globulin subfractions in serum-producing horses, with no statistical significance. We also observed that horses used to produce hyperimmune plasma developed hypoalbuminemia, although the decrease in albumin production was not statistically significant. Our findings suggest that the continuous use of horses to produce crotalid antivenom may lead to a chronic inflammatory stimulus, with changes in plasma levels of inflammatory proteins.

Evaluation of acquired passive immunity in mule foals up to 60 days of age.

Ten mule foals were used to investigate serum IgG concentration. Blood was collected from mares and foals on the day following parturition (D1), at 48 and 72 hr after birth (D2 and D3), and on D7, D14, D21, D28, D35, D42, and D60. Serum IgG concentration was determined by zinc sulfate turbidity test and was above 400 mg/dl at all time points, except for one foal on D2. Mean values were above 800 mg/dl from D1 to D7 and at D60. The difference between mares and foals at each collection time was significant, except at D1. For all other days, foal IgG values were lower than the mare IgG values. Mean IgG concentrations were lower from D14 to D42 compared with D1 and D60. The results show that the critical period for the mule foal is at D28, which can be a vulnerable window for infections. The variation pattern in IgG values is similar to that in data published for horse and donkey foals. There are no published studies to date on the subject, and the present contribution enables further understanding of the newborn mule foal.

Bartonella henselae and Bartonella clarridgeiae infection, hematological changes and associated factors in domestic cats and dogs from an Atlantic rain forest area, Brazil.

Cats are considered main reservoir of Bartonella henselae, which is transmitted to other cats especially through Ctenocephalides felis fleas, and to humans through scratching and biting. Serra da Tiririca State Park (PESET) is an Atlantic Forest area that shelters a wide variety of endemic fauna. Recently, the park has been suffering due to irregular housing construction and domestic animal population that interacts with humans and wildlife. Given that surveillance policies for animals are part of the global Strategic Framework for One Health, the aim of this study was to detect Bartonella spp. DNA in cats and dogs, evaluating laboratory changes and associated factors. Blood samples of 124 dogs and 89 cats were collected for hematology and serum chemistry analysis. DNA was extracted and tested by conventional polymerase chain reaction (PCR) targeting a fragment of the citrate synthase (gltA) gene of Bartonella spp. with specific primers. Positive samples were sequenced to identify species. Bartonella henselae and B. clarridgeiae were detected in 24.7% of cats, being, for our knowledge, the first report of B. clarridgeiae in cats from Rio de Janeiro, Brazil. None of the samples obtained from dogs tested positive in the PCR assays. No statistical significance was observed in physical and laboratory exams. We suggest that cats that inhabit PESET can be considered sources of Bartonella sp. for other cats and humans. We highlight that infected cats did not present clinical or laboratory alterations. We alert for the need of care measures, avoiding scratch and bite, particularly in immunocompromised people.

The influence of serial fecal sampling on the diagnosis of giardiasis in humans, dogs, and cats.

Giardia infection is a common clinical problem in humans and pets. The diagnosis of giardiasis is challenging as hosts intermittently excrete protozoan cysts in their feces. In the present study, we comparatively evaluated two methods of serial fecal sampling in humans, dogs, and cats from Rio de Janeiro, Brazil. The Faust et al. technique was used to examine fecal specimens collected in triplicate from 133 patients (52 humans, 60 dogs, and 21 cats). Specimens from 74 patients were received from the group assigned to carry out sampling on consecutive days - 34 humans, 35 dogs, and 5 cats, and specimens from 59 patients were received from the group assigned to carry out sampling on non-consecutive, separate days - 18 human beings, 25 dogs, and 16 cats. G. duodenalis cysts were found in stools of 30 individuals. Multiple stool sampling resulted in an increase in the number of samples that were positive for Giardia in both groups. The authors therefore conclude that multiple stool sampling increases the sensitivity of the Faust et al . technique to detect G. duodenalis cysts in samples from humans, cats and dogs.

Behavior of red cell distribution width (RDW) in a population of anemic and non-anemic dogs from an ambulatory care unit

O objetivo deste estudo foi avaliar o comportamento do índice de anisocitose na interpretação do hemograma de cães anêmicos e não anêmicos. Atualmente, contadores hematológicos automatizados veterinários fornecem alguns índices que não são calculados quando o hemograma é confeccionado manualmente. A saber: RDW-CV (Amplitude de distribuição de hemácias como coeficiente de variação) e RDW-SD (Amplitude de distribuição de hemácias como desvio-padrão). Tais índices levaram a uma nova abordagem do paciente com anemia permitindo que a anisocitose fosse mensurada de uma forma quantitativa. Foram avaliados 454 hemogramas processados no período de setembro de 2009 a março de 2011. Foram calculados média e desvio padrão e uma análise de variância foi realizada. Os cães anêmicos (125/454) apresentaram altos valores de RDW enquanto que, em animais não anêmicos, esse índice permaneceu dentro dos valores de normalidade previamente estabelecidos. O índice também estava alto em animais com anisocitose observada à microscopia. Houve uma diferença significante entre os índices dos animais que apresentaram Volume Globular Médio (VGM) elevado e normal. Concluiu-se que os índices RDW-CV e RDW-SD foram sensíveis a pequenas variações no tamanho das hemácias e na heterogeneidade, sendo mais precisos que a observação microscópica para se detectar anisocitose e mais sensível que VGM para variação eritrocitária.

The aim of this study was to evaluate the behavior of the red cell distribution width (RDW) in the interpretation of the Complete Blood Count (CBC) from anemic and non-anemic dogs. Currently, automated veterinary blood cell analyzers provide hematologic indices not calculated in manually performed CBC, such as Red Blood Cell Distribution Width measured by Variation Coefficients (RDW-CV) and Red Blood Cell Distribution Width measured by Standard Deviation (RDW-SD), which has led to new of September 2009 to March 2011. Mean and standard deviation were calculated and variance analysis was performed. Anemic dogs (125/454) presented higher values for RDW indices. In non-anemic animals, these indices were in accordance with the normal values previously established. Both indices were higher in dogs with microscopic observation of anisocytosis. There was a significant difference among índices of the animals which showed normal and increased Mean Corpuscular Volume (MCV). We concluded that RDW-CV and RDW-SD indices were sensitive to small variations on erythrocyte size and heterogeneity, being more accurate than microscopic observations to detect anisocytosis and more sensitive than MCV to erythrocyte variation.

Genetic diversity of Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil.

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

Genetic diversity of Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil / Diversidade genética de cepas de Ehrlichia canis encontradas em cães naturalmente infectados no Rio de Janeiro, Brasil

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of 1the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

O objetivo deste estudo foi caracterizar as cepas de Ehrlichia canis em cães naturalmente infectados no Rio de Janeiro, Brasil. Além disso, os achados clínicos e hematológicos observados nos cães foram relatados. O gene 16S rRNA foi utilizado como alvo da PCR para fins diagnósticos, e os genes TRP19 e TRP36 para avaliar a diversidade genética. Quinze amostras foram positivas para E. canis. PCR para o gene TRP19 produziu 11 amplicons (11/15) que foram clonados no pGEM-T easy vector para sequenciamento. A comparação das sequências completas do gene TRP19 com outras sequências depositadas no GenBank revelou uma alta identidade. Duas amostras (56C e 70C) após o ensaio da PCR, tendo como alvo o gene TRP36, geraram sequências, e a análise filogenética mostrou que a cepa 56C foi agrupada com a cepa Cuiabá 16, que é uma cepa híbrida, formada pelo genogrupo Brasileiro e o genogrupo US; e a cepa 70C agrupou com as outras cepas do genogrupo US, sugerindo a existência de pelo menos dois genogrupos de E. canis no Rio de Janeiro (US e Brasileiro). Esses animais apresentaram manifestações clínicas e hematológicas distintas, e diferentes genótipos podem expressar novos fenótipos, resultando em diferentes formas de apresentação da doença e fazendo com que o diagnóstico seja mais complexo.

Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a brazilian domestic cat (Felis catus) / Co-infecção por Cytauxzoon felis e 'Candidatus Mycoplasma haemominutum' em um gato doméstico (Felis catus) no Brasil

This article describes the first detection of Cytauxzoon felis, using molecular techniques, in a naturally infected domestic cat from Brazil, South America. Coinfection with 'Candidatus Mycoplasma haemominutum' was also found. The molecular identification of the piroplasmid species was performed by Polymerase Chain Reaction (PCR) and sequencing analysis. A 284 pb fragment of the gene encoding the 18S ribosomal RNA region was amplified and showed 99% identity with other C. felis strains from North America. In addition, PCR-RFLP (restriction fragment length polymorphism) analysis, which amplifies a 595 bp fragment of the gene encoding 16S ribosomal RNA of some bacterial species, identified the co-infecting species as 'Candidatus M. haemominutum'.

Este artigo descreve a primeira detecção de Cytauxzoon felis em um gato doméstico naturalmente infectado no Brasil, América do Sul, através de técnicas moleculares. Também foi encontrada co-infecção com 'Candidatus Mycoplasma haemominutum'. A detecção molecular da espécie do piroplasmídeo foi realizada através da reação em cadeia pela polimerase (PCR) e sequenciamento. Um fragmento de 284 pb do gene codificador da região 18S do RNA ribossomal do parasito foi sequenciada e mostrou 99% de identidade com outros isolados de C. felis da América do Norte. Ademais, através da análise por meio de PCR-RFLP (Polimorfismo no comprimento de fragmentos de restrição), que amplifica um fragmento de 595 pb do gene codificador da porção 16 do RNA ribossomal de algumas espécies de bactérias, concluiu-se que a espécie com-infectante era 'Candidatus M. haemominutum'.

A review of the occurrence of hemoplasmas (hemotrophic mycoplasmas) in Brazil.

Recent studies have been conducted in Brazil using molecular techniques for the detection of hemotrophic mycoplasmas in several mammals. In domestic cats, Mycoplasma haemofelis, "Candidatus M. haemominutum", and "Candidatus M. turicensis" infections have been identified. These species have also been found in free-ranging and captive neotropical felid species. Two canine hemoplasmas, Mycoplasma haemocanis and "Candidatus Mycoplasma haematoparvum", have been identified in dogs. In commercial swine populations, Mycoplasma suis was found to be highly prevalent, especially in sows. Moreover, novel mycoplasma species have been identified in Brazilian commercial pigs and domestic dogs. A hemoplasma infection in a human patient infected with the human immunodeficiency virus (HIV) was also recently documented. In conclusion, hemoplasma species are common and important infectious agents in Brazil. Further studies should be conducted to better understand their impact on pets, production animals, and wildlife fauna, as well as their role as zoonotic agents, particularly in immunocompromised patients.

A review of the occurrence of hemoplasmas (hemotrophic mycoplasmas) in Brazil / Uma revisão da ocorrência dos hemoplasmas (micoplasmas hemotróficos) no Brasil

Recent studies have been conducted in Brazil using molecular techniques for the detection of hemotrophic mycoplasmas in several mammals. In domestic cats, Mycoplasma haemofelis, 'Candidatus M. haemominutum', and 'Candidatus M. turicensis' infections have been identified. These species have also been found in free-ranging and captive neotropical felid species. Two canine hemoplasmas, Mycoplasma haemocanis and 'Candidatus Mycoplasma haematoparvum', have been identified in dogs. In commercial swine populations, Mycoplasma suis was found to be highly prevalent, especially in sows. Moreover, novel mycoplasma species have been identified in Brazilian commercial pigs and domestic dogs. A hemoplasma infection in a human patient infected with the human immunodeficiency virus (HIV) was also recently documented. In conclusion, hemoplasma species are common and important infectious agents in Brazil. Further studies should be conducted to better understand their impact on pets, production animals, and wildlife fauna, as well as their role as zoonotic agents, particularly in immunocompromised patients.

Estudos recentes utilizando técnicas moleculares para a detecção de micoplasmas hemotróficos em diferentes mamíferos têm sido conduzidos no Brasil. Em gatos domésticos, infecções por Mycoplasma haemofelis, 'Candidatus M. haemominutum' e 'Candidatus M. turicensis' foram identificadas. Estas espécies também foram encontradas em felídeos neotropicais de vida livre e de cativeiro. Dois hemoplasmas caninos, Mycoplasma haemocanis e 'Candidatus Mycoplasma haematoparvum', foram identificados em cães domésticos. Em populações comerciais de suínos, Mycoplasma suis possui alta prevalência, especialmente em porcas. Além disso, novas espécies de hemoplasmas foram detectadas em suínos comercias e cães. Infecção por um hemoplasma em um paciente humano infectado com o vírus da imunodeficiência humana (HIV) foi recentemente documentada. Em conclusão, espécies de hemoplasmas são comuns e importantes agentes de infecções no Brasil. Estudos futuros devem ser conduzidos para melhor entender seu impacto em cães e gatos, animais de produção e na fauna silvestre, e também para determinar o seu papel como agentes zoonóticos, particularmente em pacientes imunocomprometidos.

Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil.

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.