Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus)

ABSTRACT: This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.

RESUMO: Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)

Morphological, microbiological and ultrastructural aspects of sepsis by Aeromonas hydrophila in Piaractus mesopotamicus.

Aeromonas bacteria can cause an infection characterized by septicemia and is one of the most common pathogens in tropical fish. This disease is responsible for high morbidity and mortality rates, causing considerable losses in aquaculture. Thus, the understanding of its pathophysiology is crucial to develop control strategies of this bacterial infection in farmed fish. This study aimed to characterize early pathological aspects of acute sepsis in pacu (Piaractus mesopotamicus) experimentally infected with Aeromonas hydrophila. A total of 160 juvenile pacus were inoculated intraperitoneally with A. hydrophila (1.78 x 109 CFU/mL) and at 0 (control), 1, 3, 6, and 9 hours post-inoculation (hpi), animals were anesthetized and samples were collected for microbiological, light microscopy and transmission electron microscopy (TEM) analyzes. The results showed the occurrence of hemodynamic alterations, such as hemorrhage and congestion, which were observed mainly after 6 and 9 hpi. It was possible to re-isolate Aeromonas at all sampling times except in control group. However, just after 9 hpi it was possible to find the bacteria in all fish and tissues. Light microscopy analyses revealed a degenerative process, necrosis and vascular damage mainly at 6 and 9 hpi. According to the ultrastructural examination, areas of cellular death were identified in all examined tissues, especially at 6 and 9 hpi. However, the most severe, related to necrosis, were observed after 6 and 9 hpi. The findings suggested that this bacterium spreads in the first hpi through the fish organs, mainly affecting spleen, liver and kidney, causing irreversible lesions at the molecular level.

Feed processing effects on digestibility, palatability, excreta fermentation products and blood parameters in blue-fronted amazon parrots (Amazona aestiva)†.

Captive parrots show a high incidence of obesity and other metabolic disorders due to the consumption of unbalanced diets. Therefore, this study evaluated the digestibility and metabolic effects of transitioning blue-fronted amazons from a high fat diet (sunflower seeds) to processed diets with three degrees of starch gelatinization (SG). The same feed formulation was processed to obtain pelletized feed (PEL) at 27.1% SG; low-cooked extruded feed (EXTL ) at 81.6% SG; and high-cooked extruded feed (EXTH ) at 98.5% SG. Thirty adult parrots were fed sunflower seeds for 90 days, then were distributed in a completely randomized design with 10 repetitions per treatment, and fed one of the three prepared diets for 160 days. Feed palatability, apparent digestibility, excreta concentrations of volatile fatty acids, lactate and ammonia, initial and final radiographic examinations, blood cell counts and glucose, triglycerides, cholesterol, total protein, albumin, aspartate aminotransferase (AST) and uric acid levels were evaluated. The data were analysed by an analysis of variance and compared by Tukey's test (p < 0.05). Sunflower seed was more digestible than processed feeds (p < 0.05). Diet processing interfered with fat and starch digestibility (p < 0.001), being higher in the PEL than in the EXTH and EXTL respectively. Transitioning from sunflower seeds to balanced diets reduced serum glucose, triglycerides, cholesterol and AST (p < 0.05) and increased red blood cell, haemoglobin, lymphocyte, monocyte and leucocyte counts (p < 0.01). Radiographs indicated a decreased hourglass (p = 0.015) and a reduced heart-liver ratio after ingesting the processed feeds (p < 0.05). Feed processing did not affect blood cell counts, serum biochemistry or radiographic examinations. In conclusion, parrots preferred the extruded diet and did not require an extensive SG to properly digest the feed. Consuming the processed diets improved the birds' metabolism and health.

Linearidade e carryover do contador hematológico Horiba ® Abx Micros ESV 60 / Linearity and carryover of automated hematology counter Horiba ® Abx Micros ESV 60

O contador automático hematológico ABX Micros ESX 60 (Horiba Medical 2012) é analisador hematológico veterinário multi-espécie que realiza 50 contagens por hora, libera 18 parâmetros sanguíneos, além de fazer representações gráficas (histogramas) para leucócitos, hemácias e plaquetas. O objetivo deste trabalho é avaliar o desempenho do referido aparelho em relação à linearidade e carryover, através de controle comercial e de amostras de sangue provenientes da rotina do Laboratório de Patologia Clínica Veterinária. De acordo com resultados é possível afirmar que o presente aparelho possui um excelente coeficiente de linearidade (r2=0,99) nos parâmetros de leucócitos, eritrócitos e plaquetas em relação às diluições estudadas. Em relação aos carryover houve excelente desempenho do aparelho, contudo, houve valores não conformes nos parâmetros de CHCM e VPM em uma das metodologias realizadas que pode ser justificada pela limitação da fórmula que não considera a características do equipamento.(AU)

The automated hematology counter ABX Micros 60 ESX (Horiba Medical 2012) is veterinary hematology analyzer multi-species that carries 50 counts per hour releases 18 blood parameters, in addition to graphical representations (histograms) for leukocytes, erythrocytes and platelets. The objective of this study is to evaluate the performance of the apparatus with respect to linearity and carryover through commercial control and blood samples from the routine of Veterinary Clinical Pathology Laboratory. According to results we can say that this device has excellent linearity coefficient (r2=0.99) in leukocyte parameters, erythrocytes and platelets during that time dilutions. Regarding the carryover was excellent device performance, however, was not in conformity values ​​in the parameters of MCHC and VPM in one of the methodologies made that can be justified by the limited formula that does not consider the equipment characteristics.(AU)