Efeitos da ocratoxina no desempenho do camarão-branco-do-pacífico (Litopenaeus vannamei, Bonne) / The effects of ochratoxina in performance pacific white shrimp (Litopenaeus vannamei Bonne)

A ocratoxina é um dos maiores grupos de micotoxinas; são metabólitos secundários produzidos principalmente por fungos dos gêneros Aspergillus e Penicillium. Possui propriedades tóxicas e nefrotóxicas, está relacionada à nefropatia endêmica dos Bálcãs, a tumores do trato urinário e foi classificada pela Agência Internacional de Pesquisa do Câncer (IARC) como pertencente ao grupo 2B, por ser possivelmente carcinogênica para humanos. O objetivo do presente estudo foi avaliar os efeitos da ocratoxina A (OTA) no desempenho do camarão-branco-do-pacífico (Litopenaues vannamei). O experimento foi feito simulando o manejo produtivo de uma fazenda de camarão marinho do litoral localizada em Luís Correia, Piauí. Foram utilizados cinco tratamentos com diferentes níveis de micotoxinas: T1- 100µg/kg de OTA; T2- 500µg/kg de OTA; T3- 1000µg/kg de OTA; T4- 100µg/kg de OTA e 500µg/kg afatoxina B1 e T5 - 0,0µg/kg de OTA. A produção de OTA foi realizada por meio da fermentação do milho, utilizando-se a cepa de Aspergillus ochraceus. Rações comerciais foram contaminadas com os núcleos de milho. A detecção e a quantificação de OTA dos núcleos, das rações comerciais e dos tecidos do camarão (cefalotórax e abdome) foram realizadas por cromatografia líquida de alta eficiência (CLAE). Para simular o sistema de criação da fazenda, os animais foram cultivados por um período de oito semanas, sendo 20 animais por caixa, recebendo alimentação duas vezes por dia. O menor ganho de peso observado foi no T2 e no T4 e os maiores ganhos de peso foram obtidos no T1 e no T5, que também apresentaram a melhor conversão alimentar. Após 56 dias de experimento, foi detectada OTA residual nas amostras de abdome apenas nos camarões do T1. Logo, camarões alimentados com rações contaminadas com OTA têm seu desempenho produtivo comprometido, o que gera impactos econômicos negativos para a indústria carcinicultora, além de ser um risco à saúde do consumidor, devido aos resíduos em sua musculatura.(AU)

Ochratoxin A is the second largest group of mycotoxins. It is a secondary metabolite produced mainly by fungi of the genera Aspergillus and Penicillium, and has toxic and nephrotoxic properties that are associated with the Balkan endemic nephropathy and urinary tract tumors. The International Agency for Research on Cancer (IARC) classifies it as group 2B: possibly carcinogenic to humans. The aim of this study was to evaluate the effects of ochratoxin A (OTA) on the performance of the Pacific white shrimp (Litopenaues vannamei). The experiment simulated the productive management of a shrimp farm located on the marine coast of Luis Correia, Piauí State. Five treatments with different levels of mycotoxins were used: T1- 100µg/kg of OTA; T2- 500µg/kg of OTA; T3 - 1000µg/kg of OTA; T4- 100µg/kg of OTA and 500µg/kg afatoxina B1 and T5 - 0.0µg/kg of OTA. OTA was produced by fermenting corn, using the Aspergillus ochraceus strain. Commercial feeds were contaminated with the corn kernels. OTA in the kernels, commercial feeds and shrimp tissues (cephalothorax and abdomen) were detected and quantified via high performance liquid chromatography (HPLC). To simulate the farming system, totaling 20 animals per box. The animals were fed twice a day and raised under these conditions for eight weeks. The shrimp gained weight during the weeks of the test, when subjected to different OTA treatments. The lowest weight gain was observed in T2 and T4 and the highest weight gains were in T1 and T5, which also presented the best feed conversion ratio. After 56 days, residual OTA was detected in samples of shrimp abdomen only in T1. Therefore, the productive performance of shrimp that are fed with OTA-contaminated feed is compromised, which has a negative economic impact on the shrimp industry, and is a health risk to consumers due to residues in the muscles.(AU)

Toxicidade subaguda do extrato etanólico das folhas de Myracrodruon urundeuva sobre o ciclo estral de ratas Wistar / Subacute toxicity of Myracrodruon urundeuva leave ethanolic extract and its effects on the estrous cycle of female Wistar rats

RESUMO A fitoterapia, abrangendo o uso popular e empírico de plantas medicinais no combate e prevenção de doenças, deve ser aliada a estudos científicos que comprovem a eficácia e segurança desses compostos. No Brasil, a ampla biodiversidade vegetal da região nordeste favorece o uso e estudo de plantas com potenciais terapêuticos. A Aroeira do Sertão (Myracrodruon urundeuva Allem.) é uma espécie pertencente à família Anacardiaceae, comum no semiárido, desde o Piauí até Minas Gerais. Seu uso é diverso e disseminado por todo o País, sendo indispensáveis estudos de suas potencialidades e riscos. Devido à ampla utilização da Myracrodruon Urundeuva de forma empírica, este trabalho tem por objetivo a pesquisa de efeitos tóxicos com doses repetidas do extrato etanólico de Myracrodruon urundeuva Allem. (EEMU) e sua influência no ciclo estral de ratas Wistar. No protocolo de toxicidade subaguda foram utilizadas 25 ratas Wistar, divididas em 5 grupos (n=5), tratadas com diferentes doses do EEMU (125, 250, 500 e 1000 mg/kg) e água destilada (controle) por um período de 28 dias. Nesse período foram realizadas mensurações do consumo de água e ração e avaliação comportamental. Esses animais foram avaliados diariamente quanto a fase do ciclo estral, por meio de esfregaço vaginal a fresco, sendo observada a frequência de cada fase bem como o intervalo interestro. As ratas foram anestesiadas e o sangue foi coletado para a realização dos ensaios bioquímicos. Em seguida, foram eutanasiadas para coleta e avaliação dos órgãos internos. Durante o período de avaliação, não foram observadas alterações de comportamento, nem de consumo de água ou ração. A evolução ponderal dos animais não diferiu entre os grupos tratados. A concentração sérica de ALT foi maior nos animais tratados com EEMU 1000 mg/kg. As ratas tratadas com o extrato não apresentaram alterações significativas na frequência das fases do ciclo estral bem como duração do estro e intervalo entre estros, quando comparadas ao grupo controle. Os resultados obtidos neste estudo não apontam alterações tóxicas significativas, sistêmicas ou sobre o ciclo estral, de ratas Wistar tratadas com o extrato, nas doses avaliadas, por um período de 28 dias.

ABSTRACT The herbal medicine, comprising the popular and empirical use of medicinal plants in combating and preventing diseases, should be combined with scientific studies that prove the effectiveness and safety of these compounds. In Brazil, the vast plant biodiversity of the Northeast favors the use and study of plants with therapeutic potential. Aroeira (Myracrodruon urundeuva Allem.) is a species of the Anacardiaceae family, common in the semiarid region, from Piauí to Minas Gerais. Its use is diverse and widespread throughout the country, making studies of its potential and risks necessary. Due to the wide use of Myracrodruon urundeuva in an empirical form, this paper aims to research toxic effects with repeated doses of Myracrodruon urundeuva Allem.ethanol extract (EEMU) and its influence on the estrous cycle of female Wistar rats. In a subacute toxicity protocol 35 female Wistar rats were used, divided into 5 groups (n=5) treated with different doses of EEMU (125, 250, 500 and 1000 mg/kg) and distilled water (control) for 28 days. In this period measurements of water intake, feed consumption were carried out and a behavioral assessment was performed. The estrous cycles of these animals were evaluated daily by fresh vaginal smear, it the frequency of each phase as well as the inter-estrus intervals were observed. The rats were anesthetized and blood was collected for the biochemical assays. Then they were euthanized for the collection and evaluation of internal organs. During the evaluation period, there were no observed behavioral changes nor were the water or food consumption variations. The weight gain of the animals did not differ between treatment groups. The serum ALT was higher in animals treated with EEMU 1000 mg/kg. The rats treated with the extract showed no significant changes in the frequency of the phases of the estrous cycle, estrus duration and estrus interval when compared to the control group. The results of this study do not indicate significant toxic alterations, neither systemic nor on the estrous cycle in female rats treated with the extract, at the evaluated doses, over a period of 28 days.

Anxiolytic-like, stimulant and neuroprotective effects of Ilex paraguariensis extracts in mice.

Yerba-mate (Ilex paraguariensis St. Hil.) is the most used beverage in Latin America with approximately 426 thousand of tons consumed per year. Considering the broad use of this plant, we aimed to investigate the anxiety-like and stimulant activity of both the hydroethanolic (HE) and aqueous (AE) extracts from leaves of I. paraguariensis. Swiss mice were treated with I. paraguariensis HE or AE chronically or acutely, respectively, followed by evaluation in the elevated plus-maze (EPM; anxiety-like paradigm), open field (OF; locomotor activity) or the step-down avoidance task (memory assessment). Following behavioral protocols the brains were collected for evaluation of acetylcholinesterase (AChE) activity ex vivo. Chronic treatment with HE induced an anxiolytic-like effect and increased motor activity besides augmented AChE activity. Additionally, acute treatment with AE prevented the scopolamine-induced memory deficit in the step-down avoidance task. Overall, our results indicate the importance of the I. paraguariensis-induced CNS effects, since it is a widely used nutraceutical. We have reported anxiolytic, stimulant and neuroprotective effects for this plant species. These effects are potentially modulated by the cholinergic system as well as by caffeine.

Toxicidade reprodutiva do extrato hidroalcóolico de Samanea tubulosa Benth em ratas wistar / Reproductive toxicity study in Wistar rats treated with hydroalcoholic extract of the pods of bordão-de-velho (Samanea tubulosa Benth)

RESUMOO objetivo deste trabalho foi avaliar possíveis efeitos do extrato das vargens de bordão-de-velho (Samanea tubulosa benth) sobre a reprodução em ratas Wistar. O extrato foi preparado a partir da secagem e trituração das vagens, seguida por maceração com etanol e, posteriormente, concentrado em rotavapor. Ratas Wistar gestantes foram tratadas com 75 mg.100 g-1 de peso corporal de extrato de S. tubulosa e o grupo controle recebeu 1ml/100 g-1 de peso corporal de solução salina por gavagem durante 19 dias. Após eutanásia no 20º dia, as seguintes variáveis foram analisadas: peso corporal fetal, peso dos fetos e placentas, número de corpos lúteos e de reabsorções, implantação, fetos vivos e mortos. O extrato de vagens de S. tubulosa apresenta toxicidade para o feto causando malformações, reabsorções fetais e diminuição pronunciada do peso ao nascer. Contudo, o extrato de vagens de S. tubulosanão influencia no número de corpos lúteos e número de implantações.

ABSTRACTThe aim of this study was to evaluate possible effects of pods from bordão-de-velho (Samanea tubulosa benth) extract on the reproduction of female rats.The extract was prepared by maceration wtith ethanol of dried and crushed pods, then it was concentrated in rotary evaporator. The pregnant Wistar rats were treated with a dose of 75 mg.100 g-1of body weight of Samanea tubulosa extract, by gavage , whereas the control group received 1ml.100g-1of saline by the same route for 19 days. After the euthanasia of the animals at day 20th,the following variables were analyzed: fetal body weight, weight of fetuses and placentas, number of corpora lutea, resorption points, live and dead fetuses and number of embryonic implantations. The extract from S. tubulosa pods, at the dose studied, showed toxicity causing fetal abnormalities, fetal resorption and pronounced decrease in the weight of offspring at birth. On the other hand, , the treatment with extract of S.tubulosa pods had no influence over corpora lutea and implantation numbers.

Inhibition of angiotensin-converting enzyme increases oestradiol production in ewes submitted to oestrous synchronization protocol.

This study aimed at evaluating the effects of angiotensin-converting enzyme inhibitor (enalapril) and angiotensin II antagonist (valsartan) on the oestradiol and progesterone production in ewes submitted to oestrous synchronization protocol. The animals were weighed and randomly divided into three groups (n = 7). A pre-experiment conducted to verify the effectiveness and toxicity of enalapril (0.5 mg/kg LW) and valsartan (2.2 mg/kg LW) showed that, in the doses used, these drugs were effective in reducing blood pressure without producing toxic effects. In the experiment, all animals were subjected to oestrous synchronization protocol during 12 days. On D10, D11 and D12, animals received saline, enalapril or valsartan (same doses of the pre-experiment), according to the group randomly divided. The hormonal analysis showed an increase in oestradiol on the last day of the protocol (D12) in animals that received enalapril (p < 0.05), but not in other groups, without changing the concentration of progesterone in any of the treatments. It is concluded that valsartan and enalapril are safe and effective subcutaneously for use in sheep and that the angiotensin-converting enzyme (ACE) inhibition with enalapril leads to an increase in oestradiol production near ovulation without changing the concentration of progesterone. This shows that ACE inhibition may be a useful tool in reproductive biotechnologies involving induction and synchronization of oestrus and ovulation in sheep.

Influence of vitrification techniques and solutions on the morphology and survival of preantral follicles after in vitro culture of caprine ovarian tissue.

The objective was to compare the efficiency of various vitrification techniques and solutions for preserving morphology and viability of preantral caprine follicles enclosed in ovarian tissue. Fragments of ovarian cortex were cryopreserved by conventional vitrification (CV) in French straws, vitrification in macrotubes (MTV), or solid-surface vitrification (SSV). Six solutions containing 6 M ethylene glycol, with or without sucrose (SUC; 0.25 or 0.50 M) and/or 10% fetal calf serum (FCS) were tested (Experiment I). After 1 wk, samples were warmed and preantral follicles were examined histologically. To evaluate follicular viability (Experiment II), ovarian fragments were vitrified with the three techniques listed above, in a solution containing 0.25 M SUC and 10% FCS. After warming, follicles were assessed by the trypan blue dye exclusion test. In Experiment III, preantral follicles enclosed in ovarian tissue were vitrified using the protocol which yielded the highest percentage of viable preantral follicles (SSV with 0.25 M SUC and 10% SFB). After warming, the preantral follicles enclosed in ovarian tissue were cultured in vitro and then, were analyzed by histology and fluorescence microscopy (calcein-AM and ethidium homodimer-1). Every vitrification protocol significantly reduced the percentages of morphologically normal follicles relative to the control (88.0%); however, the addition of 0.25 M SUC and 10% FCS to the vitrification solution improved preservation of follicular morphology (67.4, 67.4, and 72.0% for CV, MTV, and SSV, respectively). Although follicular viability after SSV (80.7%) did not differ from that in fresh (non-vitrified) ovarian tissues (88.0%), after in vitro culture, percentages of viable follicles were significantly reduced (70.0%). Percentages of morphologically normal follicles after in vitro culture of vitrified ovarian tissue were similar (76.0%) to those in ovarian cortex fragments cultured without previous vitrification (83.2%). In conclusion, SSV using a solution containing 0.25 M SUC and 10% FCS, was the most efficient method for vitrifying caprine ovarian tissue.

Characterization of potentially virulent non-O1/non-O139 Vibrio cholerae strains isolated from human patients.

Traditional methods of typing Vibrio cholerae define virulent strains according to their recognition by sera directed against the known epidemic serogroups O1 and O139, overlooking potentially virulent non-O1/non-O139 strains. Here, we have undertaken the characterization of eight clinical isolates of non-O1/non-O139 V. cholerae, collected during cholera outbreaks in Brazil. Seven of these were typed as O26 and one, 17155, was defined as non-typable. A PCR-based approach has previously detected in these strains several virulence genes derived from the CTXvarphi prophage and generally associated with pathogenic strains. Here, the presence of the O1-specific wbeN gene was investigated through PCR and found to be restricted to strain 17155, as well as one of the O26 strains, 4756, although neither strain was recognized by O1-specific antisera. The same two isolates were the only strains able to express the cholera toxin in culture, assayed by western blotting. They also possessed four repeats of the heptanucleotide TTTTGAT upstream of the ctxAB genes encoding the cholera toxin. The remaining strains possessed only two intact repeats, whereas pathogenic O1 possessed four to six repeats. To define their evolutionary relationships, selected 16S-23S intergenic rRNA spacer regions were sequenced from the various strains and the resulting sequences used to build phylogenetic trees. Strains 4756 and 17155 always clustered with control O1 strains, whereas the remaining O26 strains clustered separately. These results confirm that, despite their serological phenotype, these two strains are genotypically related to O1 strains and potentially able to produce epidemic cholera.

Adaptabilidade climática de caprinos Saanen e Azul no Meio-Norte do Brasil / Climatic adaptability of Saanen and Azul goats in Brazilian Middle-North

A adaptabilidade de caprinos de dois grupos genéticos, Saanen e Azul, às condições climáticas do Meio-Norte do Brasil foi avaliada por meio dos testes de Ibéria, Benezra e Rainsby. Nos dois primeiros, foram utilizadas sete fêmeas de cada grupo racial e, no terceiro, quatro fêmeas de cada grupo. Foram realizadas quatro coletas de dados em cada período (chuvoso e seco) de 2005. Foi utilizado delineamento inteiramente ao acaso, em fatorial 2x2 (duas raças e dois períodos). Os valores do coeficiente de tolerância ao calor do teste de Ibéria no período seco diferiram entre os grupos (P<0,05) (Saanen = 97,65 e Azul = 94,31). Houve diferença entre grupos (P<0,05) quanto ao coeficiente de adaptabilidade 1 do teste de Benezra, nos dois períodos (chuvoso: Saanen = 5,13 e Azul = 3,26; seco: Saanen = 5,86 e Azul = 2,87). No teste de Rainsby, no grupo Azul, houve o retorno à temperatura de repouso nos dois períodos. Na Saanen, no período no seco, 100 minutos não foram suficientes para o retorno à temperatura de repouso. O grupo racial Azul mostrou-se mais adaptado às condições do Meio-Norte.

This study was undertaken to evaluate Saanen and Azul goats' adaptability to the Brazilian Middle-North region, based on adaptation indexes (Iberia, Benezra and Rainsby tests). A totall of seven and four females, respectively, of each group were used in two tests and four collections were performed during the rainy and dry periods of 2005 year. A completely randomized experimental design in a 2 x 2 (2 groups x 2 period) factorial treatment combination was used. Significant difference between groups (Saanen = 97.65 and Azul = 94.31) was observed for heat-tolerance coefficient (Iberia) during the dry period (P<.05). Significant differences between groups (P<0.05) were also observed for adaptability, coefficient 1 (Benezra) for both rainy (Saanen = 5.13 and Azul = 3.26) and dry periods (Saanen = 5.86 and Azul = 2.87). Based on Rainsby test, Azul goats returned to rest temperature in both periods. During the dry period 100 minutes were not enough for Saanen goats return to rest temperature. Azul goats showed higher adaptability to environment conditions of the Brazilian Middle-North region.

Mortalidade por "septicemia dos peixes tropicais" em tilápias criadas em consorciação com suínos

A rural property was observed for one year to obtain seasonal observations of Edwardsiella septicemia in tilapias (Oreochromis sp.) from an integrated fish farm (using pig excrements as food). Fifty tilapias on summer, forty-five on autumn, fifty-five on winter and one hundred and five on spring were collected and evaluated microbiologically for the presence of Edwardsiella tarda. Samples of the external surface (skin, gills and fin), intestines and muscle were analyzed. Fishes were transported alive to the laboratory, and maintained for five days for observation. The external signs observed were opacity of cornea, respiratory difficulty, disordered swimming, nodules on gills, hemorrhagic lesions under the skin and fins and incoordination of the posterior part of the body. In general, after three days of observation, the tilapias died. At necropsy, ascites with the distension of the celomatic cavity, hemorrhagic and necrotic lesions in the liver, spleen and kidneys were found. Hemorrhagic enteritis was observed in the gut tube. Edwardsiella tarda isolated presented the following biochemical profile: negative urea, sucrose, phenylalanine desaminase and malonate utilization, and positive indole, methyl red, H2S, motility, glucose with gas production and lysine decarboxylase. The mortality rates were 69.9% on spring, 63.6% on winter, 48.0% on summer and 40.0% on autumn, highest rates occurring on spring and winter (P 0,05).

Com o objetivo de estudar a variação sazonal da septicemia dos peixes tropicais provocada por Edwardsiella tarda foram observadas 50 tilápias (Oreochromis sp.) no verão, 45 no outono, 55 no inverno e 105 na primavera. Os peixes doentes apresentavam opacidade de córnea, dificuldade respiratória, nado desordenado, nódulos nas brânquias e lesões hemorrágicas na pele e nadadeiras. Após três dias de observação as tilápias morriam naturalmente. Nas necrópsias foi encontrado ascite com distensão da cavidade celomática e lesões hemorrágicas e necróticas no fígado, baço e rins. No interior do tubo digestivo observou-se enterite hemorrágica. Isolou-se Edwardsiella tarda a partir de provas positivas para H2S, lisina, indol, motilidade e glicose com gás. Os índices de mortalidade foram: na primavera 69,9%, no inverno 63,6%, no verão 48,0% e no outono 40,0%.