RESUMO

Iron deficiency anemia (IDA) in humans is defined as the decrease of total hemoglobin concentration and the non-production of the adult hemoglobin subtype 2 - HbA2 (α2δ2 chains), which is considered a marker of IDA severity in humans, dosed together with the iron serum. This study aimed to determine the standard of hemoglobin types in piglets induced to experimentally IDA in the first 21 days of life (delivery to weaning). In the present study, 40 piglets born from four naïve gilts, were randomly and equally assigned among the gilts. On the third day after delivery, the groups were randomly distributed in different environments (cement and clay floors) and according to the iron supplementation (iron dextran and placebo). Erythrocyte parameters, serum iron, and hemoglobin trait were analyzed at four moments between birth and weaning days. The group of piglets that did not receive iron dextran supplementation on the third-day post-birth and were placed in the pen without soil did not present HbA2 from the seventh day onwards on the agarose electrophoretogram (pH 8.6) and this observation was correlated to decrease of serum iron (ρ: 0.156, p=0.003) when compared to the other groups' piglets that did not present iron deficiency. In the present study was possible to determine the swine hemoglobin pattern in IDA, since HbA2 was absent in piglets with IDA in comparison to the non-ferropenic groups and the correlation between the reduction of iron levels and the absence of HbA2.

A anemia por deficiência de ferro (ADF) em humanos é definida como a diminuição da concentração de hemoglobina total e a não produção da hemoglobina adulta subtipo 2 - HbA2 (cadeias α2δ2), que é considerada um marcador de gravidade de IDA em humanos, dosado em conjunto com o soro de ferro. Este estudo teve como objetivo determinar o padrão dos tipos de hemoglobina em leitões induzidos experimentalmente à IDA nos primeiros 21 dias de vida (parto ao desmame). Quarenta leitões, nascidos de quatro marrãs nulíparas, foram distribuídos aleatoriamente e igualmente entre as leitoas. No terceiro dia após o parto, os grupos foram distribuídos aleatoriamente em diferentes ambientes (piso de cimento e barro) e de acordo com a suplementação de ferro (ferro dextrano e placebo). Parâmetros eritrocitários, ferro sérico e traço de hemoglobina foram analisados em quatro momentos, entre o nascimento e o desmame. O grupo de leitões que não recebeu suplementação de ferro dextrano no terceiro dia pós-parto e foi colocado em baia sem solo não apresentou HbA2 a partir do sétimo dia no eletroforetograma de agarose (pH 8,6) e esta observação foi correlacionada com diminuição da concentração sérica ferro (ρ: 0,156, p=0,003) quando comparados aos demais grupos leitões que não apresentavam deficiência de ferro. No presente estudo foi possível determinar o padrão hemoglobinêmico suíno na IDA, uma vez que, a HbA2 estava ausente nos leitões com ADF em comparação aos grupos não ferropênicos e há correlação entre a redução dos níveis de ferro e a ausência de HbA2.

Assuntos

RESUMO

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.

Assuntos

RESUMO

ABSTRACT Purposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, keratolimbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.

RESUMO Objetivos: Investigar a reprodutibilidade intra-la­boratorial dos fenótipos clínicos e avaliar anisotropias ópticas em córneas de coelhos com deficiência de células tronco limbais. Métodos: Lesões ao limbo foram feitas no olho direito de 23 coelhos adultos da Nova Zelândia Branco, usando um protocolo altamente agressivo, que envolveu peritomia limbal em 360 graus, ceratolimbectomia, cauterização por álcali, e remoção mecânica de epitélio remanescente. Os olhos foram clinicamente avaliados por 28 dias, inclusive por citologia de impressão corneal. As córneas que manifestaram um conjunto de alterações típicas de deficiência de células tronco limbais foram coletadas e submetidas à estudos em histopatologia e em anisotropias ópticas. Córneas saudáveis foram usadas como controles. Diferenças de caminho óptico de birrefringência relacionada à organização do colágeno estromal, e dicroísmo linear relacionado à expressão e à orientação das cadeias de glicosaminoglicanos dos proteoglicanos estromais, foram quantificados por microscopia de luz polarizada. Resultados: Um olho apresentou hipópio e foi excluído do estudo. Todos os olhos estudados (n=22) apresentaram sinais de inflamação ocular. A citologia de impressão não detectou células caliciformes na superfície corneal. Doze de 22 córneas manifestaram alterações clínicas típicas de deficiência de células tronco limbais, caracterizado por defeitos epiteliais, infiltrados inflamatórios, opacidade de moderada à severa, e neovascularização. Estudos por microscopia de luz polarizada mostraram que a deficiência de células tronco limbais aumentou a diferenças de caminho óptico corneal em 24,4% (versus controles). As cór­neas com deficiência de células tronco limbais foram menos dicroicas do que as córneas controle. Conclusões: Coelhos com deficiência de células tronco limbais, para aplicações em estudos pré-clínicos, devem ser rigorosamente selecionados para assegurar homogeneidade experimental, pois há evidências de que protocolos utilizados para indução de deficiência de células tronco limbais não estão associados com boa reprodutibilidade intra-laboratorial de fenótipos clínicos. A deficiência de células tronco limbais, como induzida aqui, alterou as propriedades ópticas anisotrópicas do estroma corneal. Tais alterações são indicativas de mudanças no empacotamento de colágeno e na orientação das cadeias de glicosaminoglicanos dos proteoglicanos. Conhecimentos nessas alterações são importantes para potencializar estratégias que visam a restabelecer a integridade morfofuncional do estromal corneal acometido pela deficiência de células tronco limbais.

Assuntos

RESUMO

PURPOSES: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. METHODS: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, keratolimbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. RESULTS: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. CONCLUSIONS: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.

Assuntos

RESUMO

Camu camu, Myrciaria dubia, is an Amazon plant that presents high levels of vitamin C in its composition. Several studies in animals and humans have demonstrated their efficiency in the prevention and treatment of various diseases. However, there are no reports of its properties in fish. The aim of this study was to evaluate the effect of the oral administration of the extract of this plant in the immune parameters in Nile tilapia, Oreochromis niloticus. 400 Nile tilapia (80 ± 5 g) were randomly distributed into 20 tanks with 1500 L capacity each (20 fish/tank). After a week of adaptation to environmental conditions, it was provided a diet for 5 weeks, using different levels of inclusion of camu camu extract: 0, 50, 100, 250, and 500 mg/kg of feed. Each treatment consisted of four replicates. It was obtained 40.5 mg of vitamin C/g of camu camu pulp powder by high-performance liquid chromatography. At the end of the trial period, fish were inoculated with Aeromonas hydrophila in the swim bladder. Samples were taken after 6; 24 and 48 h of the challenge. Results revealed that fish supplemented with this herb showed significant increase (P < 0.05) in white blood cells counts in blood and exudate, burst respiratory activity, lysozyme activity, serum bactericidal activity, direct agglutination, and melanomacrophage centers count. Red blood cells count, hemoglobin, hematocrit, and biochemical profile of fish supplemented with the herb presented no statistical differences compared to control group (P > 0.05). No histopathological lesions were observed in intestine, kidney, spleen, and gills. It can be concluded that the addition of Myrciaria dubia in tilapia feed improves the immune response and the growth after 5 weeks, especially, at a dose of 500 mg/kg.

Assuntos