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2.
Sci Rep ; 10(1): 7208, 2020 04 29.
Article in English | MEDLINE | ID: mdl-32350359

ABSTRACT

This study aims to report the presence of Neorickettsia risticii DNA in blood samples from naturally infected horses in Rio de Janeiro, provide clinicopathological findings related to the infection, and report the phylogenetic diversity of the 16S rDNA of N. risticii in order to evaluate its heterogeneity. Real-time quantitative polymerase chain reaction (qPCR) was performed to investigate the presence of N. risticii in samples collected from horses (n = 187). Five positive samples were found in the molecular screening. Hypoalbuminemia and high levels of creatine kinase and lactate dehydrogenase were the predominant findings in the biochemical analysis. The sequences were similar to those of N. risticii. Phylogenetic analysis revealed genotype segregation based on the geographical distribution in the N. risticii sequence clade. Dendrograms constructed with five hypervariable regions revealed that V4 distinguished Neorickettsia at the species level and produced a phylogeny that best represented the phylogeny obtained with the complete 16S rDNA sequence. This is the first report of N. risticii DNA in the blood of Brazilian horses based on sequences deposited in GenBank. Further studies are necessary to clarify the epidemiological chain of this vector-borne parasite in order to determine and establish appropriate preventive measures in the equine trading market.


Subject(s)
Anaplasmataceae Infections , DNA, Bacterial/genetics , Horse Diseases , Horses , Neorickettsia risticii/genetics , Phylogeny , Anaplasmataceae Infections/diagnosis , Anaplasmataceae Infections/genetics , Anaplasmataceae Infections/microbiology , Anaplasmataceae Infections/veterinary , Animals , Brazil , DNA, Ribosomal/genetics , Horse Diseases/diagnosis , Horse Diseases/genetics , Horse Diseases/microbiology , Neorickettsia risticii/isolation & purification , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics
3.
Vet Clin Pathol ; 45(2): 376-80, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27228192

ABSTRACT

BACKGROUND: Collection of fluid from the lower respiratory tract (LRT) plays an important role in both the pathophysiological investigation and diagnosis of respiratory tract disease. Enzymes such as ALP are, among others, indicators of cell damage or death, type II pneumocyte proliferation, and neutrophil invasion, and have been useful as biomarkers of respiratory disease in other species. OBJECTIVES: The purpose of this study was to determine and compare tracheal wash (TW) ALP activity in healthy horses and horses with LRT inflammation (LRTI) determined by TW cytology profile. METHODS: Tracheal washes were collected from asymptomatic adult geldings to measure ALP activity. The horses were allocated to the healthy group or the group with LRT inflammation based on differential leukocyte counts of TW preparations. Horses with > 20% neutrophils and > 1% eosinophils were allocated to the LRTI group, the horses with < 20% neutrophils and < 1% eosinophils were the controls. RESULTS: Tracheal wash ALP activity, measured using a semiautomatic chemistry analyzer, was statistically significantly higher in 18 horses with LRTI (18.9 ± 11.2 × 10(3) U/L) than in healthy horses (10.3 ± 5.9 × 10(3) U/L) (P = .021). CONCLUSIONS: Determining tracheal wash ALP activity is a simple, inexpensive and safe technique that can be used to facilitate the early diagnosis of equine respiratory disease, since it is higher in asymptomatic adult horses with a TW cytology profile consistent with LRT inflammation than in healthy adult horses with a normal TW cytology profile.


Subject(s)
Alkaline Phosphatase/metabolism , Respiratory Tract Diseases/veterinary , Trachea/enzymology , Animals , Bronchoalveolar Lavage Fluid , Horses , Male , Respiratory Tract Diseases/enzymology
4.
Ciênc. rural ; 44(1): 92-96, Jan. 2014. tab
Article in Portuguese | LILACS | ID: lil-697022

ABSTRACT

A utilidade da determinação das atividades enzimáticas no trato respiratório posterior como ferramenta diagnóstica já foi demonstrada em várias espécies. Nesse contexto, este trabalho teve por objetivo determinar a atividade da Fosfatase Alcalina (FAL) no lavado broncoalveolar (LBA) de equinos da Polícia Militar do Estado do Rio de Janeiro, comparando animais sadios com portadores assintomáticos de doença inflamatória das vias aéreas (DIVA). Para tal, foram avaliados 28 animais adultos, machos, sem histórico de doença respiratória nos dois meses anteriores ao estudo, com os resultados dos exames físicos e laboratoriais (FAL sanguínea, hematócrito, leucograma, proteína total e fibrinogênio plasmáticos) dentro dos parâmetros fisiológicos. Os equinos foram divididos em dois grupos de acordo com o resultado da citologia broncoalveolar. A determinação da atividade da FAL foi realizada por meio de espectrofotometria a partir de alíquotas do sobrenadante do LBA preservadas em nitrogênio líquido. Para a estimativa do fluido epitelial pulmonar e da atividade da FAL neste, foi realizada a correção da diluição provocada pelo lavado. Os equinos com contagem diferencial de tipos celulares compatível com DIVA apresentaram atividade de FAL no LBA menor, quando comparados aos animais sadios, podendo essa dosagem ser utilizada como complementação do diagnóstico da DIVA.


The use of determining the enzymatic activities in the posterior respiratory tract as a diagnostic tool has already been demonstrated in several species. In this context, this paper aims to determine the activity of alkaline phosphatase (ALP) in the bronchoalveolar lavage (BAL) of horses from the Military Police of the State of Rio de Janeiro, comparing healthy animals with asymptomatic carriers of an inflammatory airway disease (IAD). Twenty-eight adult male animals with no history of respiratory diseases in the last two months prior to the study were studied. Physical exam and blood laboratory test results (ALP, hematocrit, leukogram, total protein and plasma fibrinogen) were within physiological parameters. The equines were separated into two groups according to the results of the bronchoalveolar cytology. The determination of ALP was done by spectrophotometry with aliquots of the supernatant of the BAL preserved in liquid nitrogen. To estimate pulmonary epithelial lining fluid and ALP activity, correction of the dilution caused by the lavage was done. The horses with a cell type differential count compatible with IAD presented a lower ALP activity in BAL when compared to healthy animals, therefore this dosage can be used as a complement in the diagnosis of IAD.

5.
Braz. j. vet. res. anim. sci ; 50(1): 68-70, 2013.
Article in Portuguese | LILACS | ID: lil-687656

ABSTRACT

The bronchoalveolar lavage (BAL) is a sensitive method to diagnose diseases of the distal portion of the lower respiratory tract and has been broadly used by numerous researchers. Cytocentrifugation is the choice cytological preparation technique, but demands specific and costly equipment. Therefore, the present paper intends to verify the applicability of the linear smear technique to evaluate BAL samples. For this, BAL samples of 30 equines were used and the cytological preparations were done by cytocentrifugation and linear smear techniques. All glass microscope slides were fixed and stained with Giemsa for the differential cell count. Regarding the effect of the preparation technique on differential counts, no significant difference in any cell type was found. The linear smear is a reliable alternative and can be recommended as a substitution to cytocentrifugation.


O lavado broncoalveolar (LBA) é um método sensível para diagnosticar doenças do trato respiratório posterior e vem sendo utilizado por diversos pesquisadores. A citocentrifugação, técnica de escolha para processar amostras citológicas de LBA, exige equipamentos específicos e caros. Por isso, este trabalho verificou a aplicabilidade da técnica de esfregaço linear para avaliação citológica do LBA. Foram utilizadas amostras de LBA de 30 equinos adultos. As preparações citológicas foram realizadas tanto por citocentrifugação quanto por esfregaço linear. Todas as lâminas foram fixadas e coradas com Giemsa para realização da contagem celular diferencial. Não foram encontradas alterações morfológicas significativas e nem diferenças estatísticas entre nenhum dos tipos celulares processados pelos dois métodos, o que permite afirmar que o método de esfregaço linear é uma alternativa segura para avaliação morfológica celular do LBA de equinos, podendo ser utilizado no lugar da citocentrifugação quando esta não estiver disponível.


Subject(s)
Animals , Respiratory Tract Diseases/pathology , Respiratory System/anatomy & histology , Cell Biology/trends , Horses , Bronchoalveolar Lavage
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