Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats.

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Braylin induces a potent vasorelaxation, involving distinct mechanisms in superior mesenteric and iliac arteries of rats.

Arterial hypertension is a risk factor for various cardiovascular and renal diseases, representing a major public health challenge. Although a wide range of treatment options are available for blood pressure control, many hypertensive individuals remain with uncontrolled hypertension. Thus, the search for new substances with antihypertensive potential becomes necessary. Coumarins, a group of polyphenolic compounds derived from plants, have attracted intense interest due to their diverse pharmacological properties, like potent antihypertensive activities. Braylin (6-methoxyseselin) is a coumarin identified in the Zanthoxylum tingoassuiba species, described as a phosphodiesterase-4 (PDE4) inhibitor. Although different coumarin compounds have been described as potent antihypertensive agents, the activity of braylin on the cardiovascular system has yet to be investigated. To investigate the vasorelaxation properties of braylin and its possible mechanisms of action, we performed in vitro studies using superior mesenteric arteries and the iliac arteries isolated from rats. In this study, we demonstrated, for the first time, that braylin induces potent vasorelaxation, involving distinct mechanisms from two different arteries, isolated from rats. A possible inhibition of phosphodiesterase, altering the cyclic adenosine monophosphate (cAMP)/cAMP-dependent protein kinase (PKA) pathway, may be correlated with the biological action of braylin in the mesenteric vessel, while in the iliac artery, the biological action of braylin may be correlated with increase of cyclic guanosine monophosphate (cGMP), followed by BKCa, Kir, and Kv channel activation. Together, these results provide evidence that braylin can represent a potential therapeutic use in preventing and treating cardiovascular diseases.

Frequência cardíaca, lactacidemia e gasto energético de equinos da raça Crioula em provas credenciadoras ao Freio de Ouro / [Heart rate, lactate concentration and energy expenditure of Creole breed horses during qualifying tests at Freio de Ouro]

Avaliações com o intuito de mensurar marcadores de eficiência na performance esportiva do cavalo Crioulo são escassas e de fundamental importância no que tange às possíveis especificidades da raça. O objetivo do presente trabalho foi avaliar e determinar os padrões de frequência cardíaca, velocidade, concentração de lactato e gasto energético de equinos da raça Crioula durante provas credenciadoras ao Freio de Ouro. Tais variáveis foram avaliadas durante a realização das etapas funcionais da competição. Observaram-se flutuações superiores da variável frequência cardíaca (FC) durante a realização das etapas de Andadura, Figura, Volta sobre Patas e Esbarradas (And/fig/VSP) (203bpm) e menores valores na etapa Paleteada II (185bpm) (P<0,05). Em relação à velocidade, o maior valor atingido foi registrado na etapa de Paleteada II (39,7km/h). A concentração de lactato sanguíneo aferida se mostrou elevada em todas as fases da competição, sendo o maior valor observado na etapa de Paleteada II (14,5mmol/L) (P<0,05) e o menor durante a etapa de Mangueira I (9,3mmol/L). Superior gasto energético foi atribuído à etapa de And/Fig/VSP (853,28kcal/kgPV/min). Portanto, todas as etapas funcionais podem ser classificadas como anaeróbias, por apresentarem concentrações de lactato sanguíneo acima de 4mmol/L, e demandam alto gasto energético pelos competidores.(AU)

Evaluations of athletic performance markers of Crioulo breed horses are scarce yet fundamentally important regarding possible unique characteristics of this breed. This study aimed to evaluate and determine heart rate, speed, blood lactate and energy expenditure patterns of Crioulo breed horses during qualifying tests in the functional phases of the "Freio de Ouro" championship. Higher values of heart rate during the phases "andadura, figura, voltas sobre patas, esbarradas" (And/Fig/VSP) (203bpm) and lower values at "Paleteada II" (185bpm) (P<0.05) were noticed. Regarding speed variable, the maximum value was registered at "Paleteada II" (39.7km/h). During all the phases, blood lactate concentration was high, with the highest value found at "Paleteada II" (14.5mm/L) and the lowest during "Mangueira I" (9.3mm/L) (P<0.05). Superior energy expenditure was noticed in the "And/Fig/VSP" phase (853.28Kcal/kgPV/min). Thus, all functional phases can be classified as anaerobic, as blood lactate concentrations remained above 4mmol/L, with high energy demand of the horses.(AU)

Folha de amendoeira como aditivo em dietas para Betta splendens / Use of almond leaf as additive in diets for Betta splendens

O objetivo do presente estudo foi avaliar o desempenho zootécnico, os parâmetrosmorfométricos, o comportamento social, a viabilidade econômica e a análise microbiológica da água de cultivo e de tecidos corpóreos de Betta splendens alimentados com folha da amendoeira Terminalia catappa (FFDA) como aditivo. Foram utilizados 28 machos de Bettas, acondicionados individualmente em aquários de 1,5L, por 50 dias. O experimento foi realizado em delineamento experimental inteiramente ao acaso, com quatro tratamentos: 0,00%; 0,25%; 0,50% e 0,75% de inclusão da FFAD e sete repetições. Ao término do experimento, foram avaliados os parâmetros: desempenho zootécnico (ganho de peso diário, consumo de ração, conversão alimentar aparente, taxa de eficiência proteica, taxa de crescimento específico e fator de condição), morfométrico (comprimento total, padrão e da cabeça, altura, índice de perfil e índice de cabeça), comportamento social, viabilidade econômica da ração, análise microbiana do conteúdo intestinal, filé e escama e análise microbiológica da água. Pela ANOVA, pelo teste de Tukey e pela regressão (P>0,05), os parâmetros: peso final, ganho de peso, comprimento padrão, comprimento total e taxa de crescimento específico foram influenciados pelos tratamentos (P<0,05), apresentando um efeito quadrático. Assim, recomenda-se o nível de 0,50% de Terminalia catappa como aditivo em dietas de Betta splendens.(AU)

The objective of this study was to evaluate the performance, morphometric parameters, social behavior, economic viability, the presence of enterobacteria in the intestinal contents and a microbiological analysis of the water culture of Betta splendens fed with almond-tree-leaf flour (ATLF) as an additive. Twenty-eight male B. splendens were individually put in 1.5 L aquariums. The experiment was done in a completely randomized experimental design with four treatments: 0.00%; 0.25%; 0.50% and 0.75%, of the ATLF, dehydrated Terminalia catappa with seven repetitions each. At the end of the experimental period, the parameters were evaluated: performance (daily weight gain, feed intake, apparent feed conversion, protein efficiency rate, specific growth rate and condition factor), morphometric (total length, standard head, height, profile index and head index), social behavior, economic viability of the feed, microbial analysis of intestinal contents, fillet and scales, and microbiological analysis of the water. Though ANAVA, Tukey test and regression analysis (P> 0.05), the parameters: final weight, weight gain, standard length, total length and specific growth rate were influenced by the treatments (P< 0.05), presenting a quadratic effect. Therefore, the 0.50% level of Terminalia catappa is recommended as additive in Betta splendens diets.(AU)

Behavioral effects of Bj-PRO-7a, a proline-rich oligopeptide from Bothrops jararaca venom.

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M1R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), α-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M1R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Reproducibility of quantitative sensory testing applied to musculoskeletal orofacial region: Site and sex differences.

BACKGROUND: This study estimated the inter-rater reliability and agreement of the somatosensory assessment performed at masseter and temporomandibular joint (TMJ) region in a group of healthy female and male participants. METHODS: Forty healthy participants (20 men and 20 women) were evaluated in two sessions by two different examiners. Cold detection threshold (CDT), warm detection threshold (WDT), thermal sensory limen (TSL), cold pain threshold (CPT), heat pain threshold (HPT), mechanical detection threshold (MDT), mechanical pain threshold (MPT), wind-up ratio (WUR) and pressure pain threshold (PPT) were assessed on the skin overlying TMJ and masseter body. Mixed ANOVA, intraclass correlation coefficients (ICC) and standard error of measurement (SEM) were applied to the data (α = 5%). Nonoverlapping 95% confidence intervals (95% CI) of ICCs were considered significantly different. RESULTS: The ICCs of 77% of all quantitative sensory testing (QST) measurements were considered fair to excellent (ICCs: 0.47-0.97), and WUR presented the lowest values. The reliability of WDT, TSL and HPT of masseter was significantly higher than TMJ, whereas the MDT reliability of TMJ was higher than masseter. In addition, the following combination of test/sites presented significantly lower ICCs for women: HPT, MDT of TMJ and MPT of both TMJ and masseter. Finally, the highest SEM values were presented for CPT and MPT. CONCLUSION: The overall somatosensory assessment of the masticatory structures performed by two examiners can be considered sufficiently reliable to discriminate participants, except WUR. Possible site and sex influences on the reproducibility parameters should be taken into account for an appropriate interpretation and clinical application of QST. SIGNIFICANCE: The test site and participant's sex can significantly influence the relative reliability and agreement of quantitative sensory testing applied to musculoskeletal orofacial region, which affect the capacity to discriminate participants and to evaluate changes over time.

Behavioral effects of Bj-PRO-7a, a proline-rich oligopeptide from Bothrops jararaca venom

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M1R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), α-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M1R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Exigência de proteína bruta para juvenis de curimatã-pacu / Crude protein requeriment to curimatã-pacu juveniles

O objetivo deste estudo foi determinar a exigência de proteína bruta (PB) de juvenis de curimatã-pacu (Prochilodus argenteus). O experimento foi conduzido por um período de 60 dias, utilizando um delineamento inteiramente ao acaso, com quatro tratamentos (dieta contendo 24,0; 28,0; 32,0 e 36,0% de PB) e cinco repetições cada, em que 100 juvenis (peso inicial de 0,33 ± 0,01g) foram distribuídos em 20 aquários experimentais com volume de 130L, sendo considerada como unidade experimental uma caixa com cinco peixes, sob recirculação de água. Avaliaram-se o desempenho produtivo, o crescimento heterogêneo, os parâmetros morfométricos, fisiológicos e a viabilidade econômica das rações em função do nível de PB. Pela análise de regressão, foi verificado um aumento linear nos parâmetros de desempenho, morfométricos e fisiológicos, com o aumento do nível proteico nas rações. Quando comparados pelo teste de Tukey, o melhor resultado (P>0,05) foi obtido com 36% de PB. Verificou-se menor heterogeneidade de crescimento dos peixes à medida que se aumentavam os níveis de PB na ração. Assim, considerando também a viabilidade econômica, recomenda-se a utilização de rações com 36% de proteína bruta para juvenis de curimatã-pacu.(AU)

The aim of this study was to determinate crude protein (CP) requirement to curimatã-pacu juveniles (Prochilodus argenteus). The experiment was conducted during 60 days in a completely randomized design, with four treatments (24.0, 28.0, 32.0 and 36.0% CP of diet) and five replications, where a hundred juvenile fishes (initial weight de 0.33 ± 0.01g), were distributed in twenty tanks (130L each). Each tank with five juveniles was considered an experimental unit, with water recirculation. Performance, heterogeneous growth, morphometric and physiological parameters, and economic viability of the rations according to the CP levels were evaluated. For the regression analysis, a linear increase in performance, heterogeneous growth and morphometric and physiological parameters with increased of the protein in rations were observed. When compared by the Tukey test (P>0.05), the best results were obtained with 36% CP. There was lower fish growth heterogeneity as the levels of CP in the diet increased. Thus, taking into consideration the economic viability it is recommended to use feed containing 36% of crude protein to curimatã-pacu juveniles.(AU)

NO production and potassium channels activation induced by Crotalus durissus cascavella underlie mesenteric artery relaxation.

Animal toxins are natural resources for pharmacological studies. The venom of Crotalus durissus cascavella (C.d. cascavella) may be a source in the bio-prospecting of new anti-hypertensive agents. The aim of this study was to investigate vascular effects of the venom of C.d. cascavella in normotensive rats. Studies were performed using isolated mesenteric artery segments and aortic endothelial cells. The cumulative administration of the venom of C.d. cascavella (0.001-30 µg/mL) on phenylephrine (Phe; 10 µM) pre-contracted rings induced a concentration-dependent vasorelaxation in the presence of vascular endothelium (Emax = 47.9 ± 5.0% n = 8), and its effect was almost abolished in the absence of endothelium (Emax = 5.8± 2.4% n = 5 (∗∗∗p < 0.001)). Tissue viability was maintained as there was no difference in the contractile capacity of rings before and after the administration of venom. The vasorelaxant effect of the venom was also abolished when arteries were pre-contracted with potassium chloride (KCl; 80 mM) (Emax = 6.4± 0.9% n = 5, ∗∗∗p < 0.001). When assessing the participation of endothelium-derived relaxing factors, it was noted that non-selective COX inhibition with indomethacin (10 µM) caused a significant reduction in the vasorelaxant effect of C.d. cascavella (*p < 0.05). When investigating the participation of NO released by endothelium, there was a significant reduction of the vasorelaxant effect of venom in rings treated with L-NAME (100 µM; Emax = 17.5± 2.2% n = 6; **p < 0.01). Similar results were noted in the presence of ODQ (10 µM), an inhibitor of soluble guanylyl cyclase (Emax = 11.2± 3.5%, n = 6) and PTIO (100 µM), a stable radical scavenger for nitric oxide (Emax = 10.77± 3.6%, n = 6). Moreover, the venom induced the release of NO by isolated aortic endothelial cells through amperometric studies. When assessing the participation of K+ channels on the vasodilatory response of the venom, tyrode solution with 20 mM of KCl caused a significant reduction in the relaxation response (p < 0.001) (Emax = 21.3 ± 8%, n = 7), as did inhibitor of delayed rectifier K+ channels (4-amynopiridine 1 mM; Emax = 9.5 ± 1.3, %, n = 5, ***p < 0.001), and vasorelaxation was almost abolished in the presence of Iberiotoxin (IbTx 100 nM). Therefore, these results suggest that the venom of C.d. cascavella induces vasorelaxation in superior mesenteric artery rings of normotensive rats in an endothelium-dependent manner. Specifically, the venom stimulates the generation of endothelium-derived relaxing factors, especially NO, and activates vascular smooth muscle hyperpolarization through K+ channels. These data illustrate that C.d. cascavella is a source of bioactive molecules and therefore has therapeutic potential in the treatment of cardiovascular diseases such as hypertension.

Composição centesimal e de ácidos graxos do músculo Longíssimus de cordeiros confinados, alimentados com dietas contendo casca de mandioca / Centesimal and fatty acid composition of the longissimus muscle of confined lambs fed diets containing cassava peels

Objetivou-se avaliar os efeitos de inclusão da casca da mandioca (CM) sobre a composição centesimal e de ácidos graxos do músculo Longíssimus de cordeiros. Foram utilizados 32 cordeiros mestiços Santa Inês, machos não castrados, peso corporal médio de 21 ± 1,5kg. Os tratamentos foram distribuídos em delineamento inteiramente ao acaso aos animais, cujo os níveis de inclusão da CM foram (0, 10, 20, 30%) baseado na matéria seca da dieta. Utilizou-se como volumoso o feno da parte aérea de mandioca, e a relação volumoso:concentrado foi de 48:52. O experimento teve duração de 70 dias. Logo após, os cordeiros passaram por um jejum sólido de 16h e, depois, foram abatidos. A carcaça foi conduzida à câmara fria, permanecendo por 24h, a uma temperatura de 4°C. Na carcaça fria, foi retirada uma amostra do músculo Longíssimus, a qual foi congelada (4°C), até o início das análises. Os resultados de umidade, cinzas e proteína não foram influenciados pela CM, observando-se efeito linear decrescente para os teores de lipídeos. Com relação à composição de ácidos graxos, houve efeito linear para o C15:0 e efeito quadrático para C14:0, C18:0 e C22:0. Nos monoinsaturados, observou-se efeito linear para C16:1, C18:1-9c e C22:1-9c e efeito quadrático para C15:1, C17:1 e C20:1; já nos poli-insaturados, foi observado efeito linear para CLA, C20:3n-6, C20:4n-6, C20:5n-3 e C22:6n-3, e quadrático para C20:2 e C18:3n-6. Assim, conclui-se que a composição centesimal e de ácidos graxos apresenta variações em razão da inclusão da CM, porém não compromete a qualidade da carne.(AU)

This study sought to evaluate the effects of the inclusion of cassava peel on the centesimal composition and fatty acids of the Longissimus muscle of lambs. We used 32 uncastrated crossbred Santa Inês lambs, with average body weight of 21 ± 1.5 kg. Treatments were in a completely randomized design, with the inclusion of cassava peel (0, 10, 20, 30%) in the diet dry matter. Hay from the cassava shoot has been used as forage and the forage:concentrate ratio was 48:52. The experiment lasted 70 days and then the lambs underwent a fast for 16 hours, were slaughtered, and their carcasses remained at a temperature of 4°C for 24 hours. In cold carcass the Longissimus muscle were taken and frozen (4°C) until the analysis. The results for moisture, ashes and protein were not influenced by cassava peel, and a decreasing linear effect was observed for the lipids levels. In composition of unsaturated fatty acids, there was a linear effect for C15:0 and a quadratic effect for C14:0, C18:0 and C22:0. Regarding the monounsaturated fatty acids, a linear effect has been observed for C16:1, C18:1-9c and C22:1-9c, and a quadratic effect for C15:1, C17:1 and C20:1. With the polyunsaturated fatty acids, a linear effect was observed for CLA, C20:3n-6, C20:4n-6, C20:5n-3 and C22:6n-3, and a quadratic effect was seen for C20:2 and C18:3n-6. Thus, it is concluded that the centesimal and fatty acid composition varies depending on the inclusion of cassava peel, however, it does not compromise the quality of the meat.(AU)

The use of Ocimum americanum essential oil against the pathogens Aeromonas hydrophila and Gyrodactylus sp. in silver catfish (Rhamdia quelen).

UNLABELLED: The bactericidal activity (minimum inhibitory concentration (MIC)-test) of Ocimum americanum (inflorescences) essential oil (OAEO) against Aeromonas hydrophila was determined in this study. Also investigated was the potential of OAEO and the main compound found in the oil (linalool) at subinhibitory concentrations to be inhibitors of haemolysis caused by Aer. hydrophila in fish erythrocytes. An in vivo experiment was conducted to evaluate the survival of fish (Rhamdia quelen) experimentally infected with Aer. hydrophila and exposed to OAEO. A second experiment was conducted to evaluate the in vitro and in vivo activity of OAEO (mix from inflorescences and leaves) against the parasite Gyrodactylus sp. The OAEO showed weak in vitro activity against Aer. hydrophila (6400 µg ml(-1) ). Subinhibitory concentrations of OAEO (100 µg ml(-1) ) inhibited haemolysis (90%) caused by Aer. hydrophila in fish erythrocytes, however, linalool did not inhibit haemolysis activity. At the low concentrations (10 and 20 mg l(-1) ) added to the water, OAEO promoted the survival of fish experimentally infected with Aer. hydrophila. Lastly, the OAEO mix (50 mg l(-1) ) was effective against Gyrodactylus sp., significantly reducing (60%) the number of parasites in the fish. SIGNIFICANCE AND IMPACT OF THE STUDY: Phytochemicals, such as essential oils (EOs) are a great source of new molecules and have shown potential to be used in aquaculture systems. However, additional studies focused on the in vivo efficacy, mode of action and identification of the active compounds are needed. This study determined the potential of Ocimum americanum EO for use against two important fish pathogens, Aeromonas hydrophila and Gyrodactylus sp., as well as providing preliminary information about the role of the main EO compound (linalool) against Aer. hydrophila virulence.

Atividades antinociceptiva e antiinflamatória do extrato etanólico de Luehea divaricata / Antinociceptive and anti-inflammatory activities of Luehea divaricate ethanol extract

RESUMO Considerando os diferentes usos etnofarmacológicos apresentados pela planta Luehea divaricata, realizou-se este estudo com o objetivo de avaliar as atividades antinociceptiva e antinflamatória do extrato etanólico de suas folhas, em modelo animal, nas dosagens de 20, 40, 80 e 160 mg/Kg, por via oral. Foram realizados os seguintes testes: contorções abdominais induzidas pelo acido acético, placa quente, formalina e edema de pata induzido por carragenina. Foram utilizados camundongos Swiss (20-25 g) para os três primeiros testes e ratos Wistar (180-250 g), para o último, divididos em seis grupos de oito animais, totalizando 48 animais em cada parâmetro de avaliação. Os resultados foram analisados estatisticamente pela análise de variância a 5% de probabilidade, para verificar quais os tratamentos que diferiram entre si, e estes foram submetidos aos testes de Kruskall-Wallis e Student-Newman-Keuls. O extrato etanólico das folhas de L. divaricata (EEtOH-Ld), nas diferentes doses estudadas, apresentou significativa atividade antinociceptiva sobre a dor induzida quimicamente por injeções intraperitoneal de acido acético e intraplantar de formalina. Na dosagem de 160 mg/Kg, esse extrato apresentou ação analgésica central, aos 120 minutos de observação, no teste de placa quente e reduziu o edema de pata induzido pela administração de carragenina, uma hora após a administração do agente inflamatório, semelhante ao efeito produzido pelo fármaco padrão.

ABSTRACT Considering the different ethnopharmacological uses submitted by the plant Luehea divaricata, this study took place in order to evaluate the antinociceptive and anti-inflammatory activities of the ethanol extract of the leaves in an animal model, the dosages of 20, 40, 80 and 160 mg/kg by oral intake. The following tests were performed: writhing induced by acetic acid, hot plate, formalin, and paw edema induced by carrageenan. Swiss mice (20-25 g) were used for the first three tests and Wistar rats (180-250 g) for the last, divided into six groups, each of eight animals, totaling 48 animals for each assessment parameter. The results were statistically analyzed by analysis of variance at 5% probability to verify which treatments differ, and these were tested by Kruskal-Wallis and Student-Newman-Keuls. The ethanol extract of L. divaricataleaves (EEtOH-Ld) at the different studied doses showed significant antinociceptive activity on chemically induced pain by intraperitoneal injections of acetic acid and intraplantar formalin. At a dosage of 160 mg/kg, this extract showed a central analgesic action after 120 minutes of observation in the hot plate test and reduced action in the paw edema induced by carrageenan one hour after the administration of the inflammatory agent, similar to the effect produced by the standard drug.

Reduction of Toxoplasma gondii Development Due to Inhibition of Parasite Antioxidant Enzymes by a Dinuclear Iron(III) Compound.

Toxoplasma gondii, the causative agent of toxoplasmosis, is an obligate intracellular protozoan that can infect a wide range of vertebrate cells. Here, we describe the cytotoxic effects of the dinuclear iron compound [Fe(HPCINOL)(SO4)]2-µ-oxo, in which HPCINOL is the ligand 1-(bis-pyridin-2-ylmethyl-amino)-3-chloropropan-2-ol, on T. gondii infecting LLC-MK2 host cells. This compound was not toxic to LLC-MK2 cells at concentrations of up to 200 µM but was very active against the parasite, with a 50% inhibitory concentration (IC50) of 3.6 µM after 48 h of treatment. Cyst formation was observed after treatment, as indicated by the appearance of a cyst wall, Dolichos biflorus lectin staining, and scanning and transmission electron microscopy characteristics. Ultrastructural changes were also seen in T. gondii, including membrane blebs and clefts in the cytoplasm, with inclusions similar to amylopectin granules, which are typically found in bradyzoites. An analysis of the cell death pathways in the parasite revealed that the compound caused a combination of apoptosis and autophagy. Fluorescence assays demonstrated that the redox environment in the LLC-MK2 cells becomes oxidant in the presence of the iron compound. Furthermore, a reduction in superoxide dismutase and catalase activities in the treated parasites and the presence of reactive oxygen species within the parasitophorous vacuoles were observed, indicating an impaired protozoan response against these radicals. These findings suggest that this compound disturbs the redox equilibrium of T. gondii, inducing cystogenesis and parasite death.

Effects of anesthesia with the essential oil of Ocimum gratissimum L. in parameters of fish stress / Efeitos da anestesia com o óleo essencial de Ocimum gratissimum L. em parâmetros de estresse de peixes

RESUMO: The effects of anesthesia with the essential oil of Ocimum gratissimum (EOO) in parameters of stress after handling were investigated in silver catfish (Rhamdia quelen). EOO was obtained from the aerial parts by hydrodistillation. Juveniles were anesthetized with 70 or 300 mg L-1 EOO and submitted to air exposure for 1 minute. The fishes were sampled immediately or transferred to anesthetic-free aquaria until sampling. In the first experiment, juveniles had their blood collected at 0, 1, 4, and 8 h after handling to assay plasma cortisol and blood glucose levels. The unanesthetized animals were restrained manually for blood collection. In the second experiment, water samples of the recovery aquaria were collected to evaluate net ion fluxes at 0 - 4 h and 4 - 8 h. Water and ethanol controls were also performed under the same conditions. The results showed that the cortisol levels did not differ among the treatments. Hyperglycemia was verified in fish exposed to 70 and 300 mg L-1 EOO at 1 h and 4 h after handling. After 8 h, cortisol and glucose concentrations were lower or similar than those from immediately after handling for all treatments. EOO anesthesia prevented Na+ efflux observed in the control groups in both flux periods. There were net Cl- and K+ effluxes at 0 - 4 h and influxes at 4 - 8 h after handling in most treatments, and these fluxes did not differ among the treatments. The results suggest that EOO did not impair stress recovery and did not act as an additional handling stressor in silver catfish.

RESUMO: Os efeitos da anestesia com o óleo essencial de Ocimum gratissimum (EOO) em parâmetros de estresse após manuseio foram investigados em jundiás (Rhamdia quelen). EOO foi obtido a partir das partes aéreas por hidrodestilação. Os juvenis foram anestesiados com 70 ou 300 mg L-1 de EOO e expostos ao ar por 1 minuto. Os peixes foram amostrados imediatamente ou transferidos para aquários sem anestésico até amostragem. No primeiro experimento, os juvenis tiveram seu sangue coletado em 0, 1, 4, e 8 h após manuseio para avaliar os níveis de cortisol e glicemia. Os animais não anestesiados foram contidos manualmente para coleta sanguínea. No segundo experimento, amostras de água foram coletadas do aquário de recuperação dos animais para avaliação do fluxo iônico entre 0 - 4 h e 4 - 8 h. Grupos controles em água e etanol também foram realizados sobre as mesmas condições. Os resultados demonstraram que os níveis de cortisol não diferiram entre os tratamentos. Hiperglicemia foi detectada em peixes expostos a 70 e 300 mg L-1 de EOO em 1 h e 4 h após o manuseio. Após 8 h, os teores de cortisol e glicose foram menores ou similares aqueles imediatamente após o manuseio para todos os tratamentos. A anestesia com EOO preveniu o efluxo de Na+ observado para os grupos controle em ambos os períodos avaliados. Ocorreram efluxos de Cl- and K+ entre 0 - 4 h e influxos entre 4 - 8 h após o manuseio para a maioria dos tratamentos, e estes eventos não diferiram entre os tratamentos. Os resultados sugerem que o EOO não prejudica a recuperação do animal frente ao evento estressor ou atua como estressor adicional ao manuseio em jundiás.