Resumo Os lactobacilos são probióticos com capacidade de desintoxicação da Aflatoxina (AF), encontrados em produtos fermentados, TGI de animais e meio ambiente. O objetivo deste estudo foi investigar a capacidade de isolados de frango de corte de Lactobacillus contra a Aflatoxina B1 (AFB1). Para tanto, 5 isolados de Lactobacillus de intestino de frango foram incubados com 100 ppb AFB1 em meio aquoso, e o efeito de diferentes parâmetros (frações celulares, tempo, temperatura, pH) na desintoxicação foi determinado por CLAE. O efeito melhorador de Lactobacillus salivarius (LS) contra AFB1 foi estudado em frangos de corte. Os resultados revelaram que LS (CR. 4) apresentou os melhores resultados (in vitro) em comparação com outros isolados [L. salivarius (CR. 3, CR. 4), L. agilis (CE. 2.1, CE. 3.1) e L. crispatus (CE. 28)]. Detritos celulares de CR. 4 mostraram desintoxicação significativamente maior (P < 0.05). A quantidade máxima de AFB1 foi desintoxicada a 30 °C (97%), pH 4.0 (99%) e 6 h (99,97%). O estudo in vivo mostrou que AFB1 diminuiu o ganho de peso (1,269 ± 0.04 g / ave), alimento consumido (2,161 ± 0.08 g / ave), proteína total sérica (2.42 ± 0.34 g / dl), albumina sérica (0.5 ± 0.22 gm / dl) e título de anticorpo (4.2 ± 0.83). As enzimas da função hepática foram encontradas (alanina transaminase (ALT): 32 ± 10.7 U / L) e aspartato transaminase (AST): 314.8 ± 27 U / L) elevadas em AFB1 alimentados com frangos. O tratamento com 1% LS não só diminuiu os efeitos tóxicos de AFB1 (grupo D), mas também melhorou a saúde geral dos frangos devido aos seus efeitos probióticos (p < 0.05) em comparação com o controle negativo (grupo A). A capacidade de desintoxicação do LS foi melhor do que o aglutinante comercial (CB) (0.2% Protmyc). Concluiu-se que a desintoxicação de AFB1 por Lactobacillus foi dependente da cepa, temperatura, pH e tempo. LS tem capacidade de desintoxicação contra AFB1 in vivo.

Abstract Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.

SUMMARY: The postmortem diagnosis of death by drowning is one of the most difficult issues in forensic pathology. We investigated possible evidence differentiating saltwater drowning from freshwater drowning by histopathological changes in brain, heart, lungs, liver, and kidneys tissues. A cross section descriptive study was carried out on eighteen 12-week-old male Wistar rats; they were divided equally into 3 groups. Group 1: control group; Group 2: death by drowning in freshwater; Group 3: death by drowning in saltwater. Immediately after death, all tested organs were removed and fixed for histopathological examination. The brain of freshwater group depicted degenerated neurocytes with dystrophic changes in the form of shrunken cell, pyknotic nuclei and deeply eosinophilic cytoplasm. The heart showed clear evidence of myocyte injuries in saltwater drowning compared to the control and freshwater groups. The kidneys of rats drown in saltwater revealed more glomerular destruction with no differences in tubulo-interstitial changes in comparison with those drown in freshwater. In the lungs, the changes in freshwater were restricted to the alveoli, and the bronchial changes were more distinctive in saltwater. No disturbed liver architecture was seen in both test groups, however hydropic degeneration, congested vessels, and sinusoids were more distinct in saltwater group. In conclusion, diagnostic differentiation between fresh and saltwater drowning was reliable in rats’ lungs and heart with minimal differentiation in liver, kidneys, and brain. Further studies of drowning with different staining techniques will help to clarify the potential role of histopathological changes in body organs as indicator of drowning.

RESUMEN: El diagnóstico post mortem de muerte por ahogamiento es uno de los temas más difíciles de la patología forense. Investigamos la posible evidencia que diferencia el ahogamiento en agua salada del ahogamiento en agua dulce por cambios histopatológicos en los tejidos del cerebro, el corazón, los pulmones, el hígado y los riñones. Se realizó un estudio descriptivo de corte transversal en dieciocho ratas Wistar macho de 12 semanas de edad; se dividieron por igual en 3 grupos. Grupo 1: grupo control; Grupo 2: muerte por ahogamiento en agua dulce; Grupo 3: muerte por ahogamiento en agua salada. Inmediatamente después de la muerte, se extirparon todos los órganos analizados y se fijaron para el examen histopatológico. El cerebro del grupo de agua dulce mostró neurocitos degenerados con cambios distróficos en forma de células encogidas, núcleos picnóticos y citoplasma profundamente eosinofílico. El corazón mostró una clara evidencia de lesiones de miocitos en los ahogamientos en agua salada en comparación con los grupos de control y de agua dulce. Los riñones de ratas ahogadas en agua salada revelaron una mayor destrucción glomerular sin diferencias en los cambios túbulo-intersticiales en comparación con las ahogadas en agua dulce. En los pulmones, los cambios en agua dulce se restringieron a los alvéolos y los cambios bronquiales fueron más distintivos en agua salada. No se observó una arquitectura hepática alterada en ambos grupos de prueba, sin embargo, la degeneración hidrópica, los vasos congestionados y los sinusoides fueron más distintos en el grupo de agua salada. En conclusión, la diferenciación diagnóstica entre ahogamiento en agua dulce y salada fue confiable en los pulmones y el corazón de las ratas con una diferenciación mínima en el hígado, los riñones y el cerebro. Estudios adicionales de ahogamiento con diferentes técnicas de tinción ayudarán a aclarar el papel potencial de los cambios histopatológicos en los órganos del cuerpo como indicador de ahogamiento.

RESUMO: Os rizóbios são bactérias que vivem no solo, caracterizadas pela capacidade única de colonizar as raízes das culturas de leguminosas, onde formam nódulos de fixação de nitrogênio. Foram acumuladas diversas evidências com o objetivo de identificar os benefícios associados ao uso de rizóbios como agentes de controle biológico contra patógenos de solo, além da fixação biológica de nitrogênio. Neste estudo, dos quatro isolados de rizóbios testados, Bradyrhizobium sp. inibiu o crescimento radial de todos os fungos de teste, a saber: Macrophomina phaseolina, Rhizoctonia solani, Fusarium solani e F. oxysporum, enquanto outros três isolados de rizóbios inibiram o crescimento de pelo menos três fungos. Os filtrados de cultura livre de células de rizóbios também apresentaram atividade nematicida, matando juvenis de segundo estágio de Meloidogyne javanica em graus variados. Os isolados de rizóbios aplicados sozinhos ou combinados com solo corrigido com torta de mostarda causaram redução significativa (p<0,05) da quantidade de fungos causadores da podridão de raízes, bem como de nematoides de galha, nas raízes da soja. As plantas tratadas com rizóbios apresentaram menor penetração de nematoides nas raízes do que as plantas de controle não tratadas. Foi observado aumento da eficácia dos rizóbios contra os nematoides no solo corrigido com bolo de mostarda a 1%. Os rizóbios também melhoraram o crescimento da soja, produzindo plantas mais altas, embora com nodulação deficiente. A planta mais alta foi encontrada no tratamento com torta de mostarda NFB-1 (1%), mas na maioria dos casos a aplicação combinada não proporcionou nenhuma vantagem adicional.

ABSTRACT: Rhizobia are soil bacteria, characterized by their unique ability to colonize the roots of leguminous crops, where they form nitrogen fixing nodules. Considerable evidence has been accumulated to identify the benefits associated with use of rhizobia as biocontrol agents against soil-borne pathogens, in addition to biological nitrogen fixation. In this study, out of four rhizobial isolates tested, Bradyrhizobium sp. inhibited the radial growth of all the test fungi viz: Macrophomina phaseolina, Rhizoctonia solani, Fusarium solani and F. oxysporum, while other 3 rhizobial isolates inhibited the growth of at least three fungi. Cell free culture filtrates of rhizobia also showed nematicidal activity by killing second-stage juveniles of Meloidogyne javanica at varying degrees. Application of rhizobial isolates alone or in mustard cake amended soil significantly (p<0.05) by suppression of root rotting fungi and root knot nematode on soybean roots. Rhizobia-treated plants showed less penetration of nematodes in roots than untreated control plants. Efficacy of rhizobia was found to increase against nematodes in mustard cake at 1% amended soil. Rhizobia also improved soybean growth by producing taller plants although plants showed poor nodulation. The tallest plant was found in some nitrogen fixing bacteria (NFB) + mustard cake (1%) treatment, but in most cases, the combined application did not offer any added advantage.

RESUMO: Foi realizada uma experiência em vasos para estudar a capacidade de nodulação de algumas espécies de leguminosas forrageiras. O cornichão (Lotus corniculatus L.), o sanfeno (Onobrychis viciifolia Scop.) e o trevo-subterrâneo (Trifolium subterraneum L.) foram cultivados em cultura isolada (100%) e em combinação com a festuca-alta (Festuca arundinacea Schreb.) nas seguintes proporções: cornichão + festuca-alta (50:50%); sanfeno + festuca-alta (50:50%); trevo-subterrâneo + festuca-alta (50:50%); cornichão + trevo-subterrâneo + festuca-alta (33:33:33%); sanfeno + trevo-subterrâneo + festuca-alta (33:33:33%). Nas culturas cultivadas isoladamente, o cornichão apresentou o maior índice de nodulação (31), seguido pelo sanfeno (26) e pelo trevo-subterrâneo (26). O índice de nodulação nas misturas binárias com gramíneas teve aumento de 16,1% para o cornichão e de 23,1% para o trevo-subterrâneo. O cornichão, isolado e em combinação, apresentou a maior eficiência do uso de fósforo para a formação de nódulos.

ABSTRACT: A pot experiment was conducted to study the nodulating capacity of some forage legumes. Birdsfoot trefoil (Lotus corniculatus L), sainfoin (Onobrychis viciifolia Scop.) and subterranean clover (Trifolium subterraneum L.) were grown in pure culture (100%) and in mixtures with tall fescue (Festuca arundinacea Schreb.) in the ratios: birdsfoot trefoil + tall fescue (50:50%); sainfoin + tall fescue (50:50%); subterranean clover + tall fescue (50:50%); birdsfoot trefoil + subterranean clover + tall fescue (33:33:33%); sainfoin + subterranean clover + tall fescue (33:33:33%). Birdsfoot trefoil from the crops cultivated in pure stands showed the highest nodulation rating (31), followed by sainfoin (26) and subterranean clover (26). Nodulation rating in binary mixtures with grass component increased to 16.1% for birdsfoot trefoil and to 23.1% for subclover. Birdsfoot trefoil, both in pure stands and in mixtures, showed the highest phosphorus use efficiency for nodule formation.

The biometric and biochemical attributes of alfalfa (Medicago sativa L.) seedlings were studied after their exposure to 0-120 µM cadmium for 28 days using hydroponic culture. The growth, photosynthetic area and pigment contents (chlorophyll a, b and total) declined significantly (p < 0.05) in the presence of high cadmium concentrations (90 and 120 µM). A steady increase in lipid peroxidation assessed via MDA production was observed with increasing levels of cadmium. Cadmium uptake by the plant tissues was concentration dependent. The roots accumulated 1020 µg g-1 of cadmium which was two folds than leaves. An enhanced production up to 600 µg g-1 of proline was observed at higher levels of cadmium. Though, cadmium toxicity was expressed in terms of decline in growth variables, chlorophyll content and oxidative damage but restricted transfer of the metal to the aerial tissue and greater production of proline in response to higher metal content seems to alleviate cadmium toxicity. Thus, ability of alfalfa plants to tolerate high cadmium concentrations can be a manifestation of effective defensive mechanism derived from differential accumulation of metal in plant tissues in addition to enhanced production of proline.