Inocuidade micotoxicológica e viabilidade de Aspergillus spp com potencial probiótico provenientes do trato digestório bovino / Mycotoxicological safety and viability of Aspergillus spp with probiotic potential from the bovine digestive tract]

Objetivou-se, com a presente pesquisa, avaliar a viabilidade de Aspergillus spp. com potencial probiótico durante o armazenamento e diante da pressão da microbiota autóctone, bem como a tolerância aos principais ácidos graxos da fermentação ruminal. Verificou-se também a inocuidade micotoxicológica desses isolados fúngicos cultivados em meio de cultivo sólido. Foram avaliados 20 isolados de Aspergillus spp., provenientes do trato gastrointestinal de bovinos criados em Urochloa decumbens lignificada. Esses fungos foram identificados por análise de sequências do DNAr e foram selecionados por apresentarem expressivo potencial celulolítico. O método vapor de amônia foi utilizado para detecção de cepas produtoras de micotoxinas. Os isolados foram avaliados quanto à viabilidade de crescimento em fluido ruminal por até 96 horas e estocagem em condições ambientais. Observou-se que os fungos avaliados não produziram aflatoxinas e que 95% dos isolados apresentaram resistência aos ácidos ruminais. Dois isolados, selecionados a partir das análises anteriores, apresentaram viabilidade sob a pressão da microbiota autóctone e de metabólitos do ecossistema ruminal e permanecem viáveis por, no mínimo, dois anos. Conclui-se que os isolados do gênero Aspergillus selecionados nesta pesquisa apresentam características fisiológicas para serem utilizados com aditivos microbianos ou probióticos para o ambiente ruminal.(AU)

The aims in this study were to evaluate the viability of Aspergillus spp. with probiotic potential during storage, pressure of autochthonous microbiota and tolerance to the main fatty acids of ruminal fermentation. The mycotoxicological safety was also verified. Twenty isolates from the gastrointestinal tract of cattle raised in lignified Urochloa decumbens were identified by rDNA sequence analysis and were previously selected because they showed significant cellulolytic potential. The ammonia vapor method was used to detect the production of mycotoxins. The isolates were evaluated for viability of ruminal fluid growth for up to 96 hours and storage under environmental conditions. The evaluated fungi did not produce aflatoxins, and 95% of them had resistance to ruminal acids. Two isolates, selected according these tests, presented viability on autochthonous microbiota pressure and metabolites from the ruminal ecosystem and remain viable for at least two years. In this research, the selected Aspergillus spp. isolates present physiological characteristics to be used with microbial additives or probiotic.(AU)

População de protozoários ruminais em novilhos zebuínos alimentados com ou sem volumoso / Population of Protozoa in zebu steers fed with or without bulk

This study aimed to quantify and identify the profile of the rumen protozoa population of beef steers fed with or without roughage. Nellore crossbred steers raised in extensive system on lignified tropical pastures with mineral supplementation and steers confined for 60 days only receiving pelletized concentrate and whole corn kernels were evaluated. After slaughter, rumen fluid was collected and one ml aliquots were diluted in nine ml formaldehyde solution at 10%. The counts of small, medium and large protozoa were held in Sedgewick Rafter chambers and identification of genus was possible after staining lugol and optical microscope with a 40X objective. The concentration of rumen protozoa positively correlated with pH ruminal. Cattle fed without roughage had significantly lower rumen protozoa population (P<0.05). Animals fed roughage had higher occurrence of Dasytrichia genus, Charonina, Entodinum, Diplodinium, Ostracodinium and Epidinium while those fed without bulk, the Buetschilia, Isotricha, Eodinium, Polyplastron, Elyplastron, Metadinium and Enoploplastron were the most frequent genus.(AU)

Efeito da inclusão da torta de macaúba na população de protozoários do rúmen de vacas leiteiras / The effect of macauba meal inclusion in the ruminal protozoa population of dairy cows

Avaliaram-se os efeitos da inclusão da torta de macaúba (TM) na dieta sobre a concentração e o perfil da população de protozoários ciliados do rúmen. Foram utilizadas oito vacas Holandesas, dispostas em dois quadrados latinos 4x4 e alimentadas com diferentes níveis de inclusão da TM (0, 100, 200 e 300g kg-1 de matéria seca na dieta total). Para quantificação e identificação dos protozoários, amostras de 1mL do líquido ruminal de cada animal foram diluídas em 9mL de solução de formaldeído a 10%. Após diluições decimais sucessivas em solução salina, os protozoários foram quantificados em câmara de Sedgewick. A identificação foi realizada sob a luz da microscopia óptica, utilizando-se a objetiva de 40x para caracterizar o mínimo de 100 indivíduos por amostra. As concentrações de protozoários pequenos (P<0,01), médios (P=0,05), grandes (P=0,01) e o número total (P<0,01) apresentaram comportamento linear decrescente com o aumento dos níveis de inclusão da TM nas dietas. Foram encontradas características morfológicas típicas de 16 gêneros de protozoários ciliados, sendo os gêneros Entodinium e Eodinium os mais frequentes no líquido ruminal dos animais avaliados. O perfil da população de protozoários ciliados sofreu pequenas alterações com a inclusão da TM, sendo observada alteração na participação percentual dos gêneros Charonina (P = 0,03) e Polyplastron (P<0,01). A utilização da TM na dieta de bovinos promove redução na concentração e modificação no perfil da população de protozoários ciliados do rúmen.

The effects of meal macaúba (MM) inclusion on the concentration and profile of the ciliate protozoa of the bovine rumen were evaluated. Eight Holstein cows were used during second lactation, with average 480 kg of body weight, arranged in two 4x4 Latin squares simultaneously and fed four diets with different concentrations of the MM (0, 100, 200 and 300 g kg-1). For identification and quantification of protozoa, samples of ruminal fluid from each animal with 1 ml were diluted in 9mL of 10% formaldehyde. After consecutive decimal dilutions in saline solution, protozoa were assessed in a Sedgewick chamber. The identification was carried out under the optical light microscope using a 40x objective to characterize at least 100 individuals per sample. The concentrations of small (P<0.01), medium (P=0.05), large (P= 0.01), and the total number (P<0.01) of protozoa decreased linearly with the increasing MM inclusion. Morphological characteristics typical of 16 genera of ciliates were found, being the Entodinium spp. and Eodinium spp. the most frequent in the rumen fluid of animals evaluated. The profile of the population of ciliated protozoa suffer minor changes with the addition of MM, with a change in the percentage share of Charonina spp. (P = 0.03) and Polyplastron spp. (P <0.01). The use of the MM in the cattle diet promotes reduction in the concentration and change in the profile of the population of rumen ciliates protozoa.

Effects of different extracts of the mushroom Agaricus blazei Murill on the hematologic profile of mice with Ehrlich tumor / Efeitos de diferentes extratos do cogumelo Agaricus blazei Murill sobre o perfil hematológico de camundongos com tumor de Ehrlich

The aim of this study was to investigate the effect of the mushroom Agaricus blazeii Murril (ABM) extracts on the hematological profile of Swiss mice bearing an Ehrlich solid tumor. Three fractions (total extract, polysaccharides, and supernatant) of ABM extracts obtained by four methods (ultrasonic or water bath, at pH 4 or pH 7) were administered to mice over 21 days. Polysaccharide solutions were analyzed by gas and liquid chromatography that showed both mannose and glucose concentrations. The method of extraction influenced the degree of glucose polymerization and the mannose/glucose relationship. The treatment with ABM supernatant at pH 7 and water bath was associated with reduced concentrations of leukocytes and lymphocytes and altered the percentage of CD4+ and CD8+ lymphocytes in Ehrlich tumor-bearing mice. The treatment with the ABM extract in water bath and ultrasound at pH 4 resulted in lower lymphocyte counts, regardless of tumor presence, and greater granulocyte values in mice with Ehrlich tumor than in controls. We concluded that different fractions and methods of extraction of A. blazei produced differing blood profiles in mice inoculated with the Ehrlich tumor.

O objetivo deste estudo foi investigar o efeito de diferentes extratos do cogumelo Agaricus blazeii Murril (ABM) sobre o perfil hematológico de camundongos Swiss portadores de tumor de Ehrlich sólido. Três frações (extrato total, polissacarídeos e sobrenadante) dos extratos de ABM foram obtidas por quatro métodos (sonificador, banho-maria, em pH 4 ou pH 7) e administradas para camundongos durante 21 dias. Soluções de polissacarídeos foram analisadas por cromatografia gasosa e líquida, que mostraram concentrações de glucose e manose. O método de extração influenciou o grau de polimerização da glicose e a relação manose/glucose. O tratamento com o sobrenadante de ABM (em pH 7 e banho-maria) estava associado com reduzidas concentrações de leucócitos e linfócitos, além de alterar a porcentagem de linfócitos CD4+ e CD8+ em camundongos portadores de tumor sólido de Ehrlich. O tratamento com extratos de ABM, obtidos tanto em banho-maria como no sonificador em pH 4, resultou nas mais baixas contagens de linfócitos, independentemente da presença do tumor, e nos maiores valores de granulócitos em camundongos com tumor de Ehrlich. Conclui-se que os diferentes métodos de extração com as respectivas frações de A. blazei são capazes de intereferir no perfil hematológico de camundongos com tumor sólido de Ehrlich.

Desenvolvimento de bezerros leiteiros alimentados com silagem de leite de transição: I - Trato digestivo / Development of dairy calves fed silage transition milk: I - Digestive tract

Avaliou-se o desenvolvimento do trato digestivo de bezerros Holandeses aleitados com silagem de leite de transição. Foram utilizados 18 animais, com peso corporal inicial médio de 36,50kg (±4,03), alimentados individualmente em baias com concentrado, feno de Cynodon sp., água e suplemento mineral oferecidos ad libitum. O delineamento experimental foi inteiramente ao acaso, com três tratamentos e seis repetições. Os tratamentos consistiram em leite integral e silagem de leite de transição diluída em água ou misturada em leite, correspondendo a quatro litros diários divididos em duas refeições, durante 59 dias. Aos 60 dias, os bezerros foram abatidos e realizaram-se pesagens do trato digestivo e medidas histológicas das papilas ruminais. O sistema de aleitamento não interferiu nos consumos de concentrado e matéria seca total e no desenvolvimento de papilas ruminais (P>0,05). A silagem de leite de transição diluída em água resultou em menor peso corporal, menor peso corporal vazio e absoluto do abomaso e maior desenvolvimento intestinal. O aleitamento com silagem misturada em leite não comprometeu o desenvolvimento corporal e do trato digestivo até 60 dias de idade, representando alternativa ao aleitamento com leite...

This study aimed to evaluate the development of the digestive tract of Holstein calves which suckled milk from silage transition. We used 18 animals, with an average initial body weight of 36.50kg (±4.03), individually housed in pens with concentrate, hay Cynodon sp., water and mineral supplement offered ad libitum. The experimental design was completely randomized with three treatments and six replications. Treatments consisted of whole milk and silage transitional milk, diluted with water or mixed with milk, corresponding to four liters per day divided over two meals during 59 days. The food intake was monitored daily and weighed every week after the supply of liquid diet in the morning. At 60 days, the calves were slaughtered and the digestive tract was weighed and histological measures of rumen papillae were taken. The nursing system did not affect the consumption of concentrate and total dry matter and the development of rumen papillae (P>0.05). The silage transition milk diluted in water resulted in lower final body weight, lower body weight and absolute emptiness abomasum and intestinal further development. Milk transition silage mixed in milk did not affect the development of the digestive tract and body until 60 days of age, representing an alternative to milk...

Desenvolvimento de bezerros leiteiros alimentados com silagem de leite de transição: II - Órgãos internos / Development of dairy calves fed silage transition milk: II - Internal organs

Avaliou-se o tamanho absoluto e relativo dos órgãos internos de bezerros Holandeses aleitados com leite, silagem de leite de transição diluída em água ou misturada em leite. Foram utilizados 18 animais, machos, com 60 dias de idade e peso corporal médio de 70,3kg (±11,8kg). O delineamento experimental foi inteiramente ao acaso, com três tratamentos e seis repetições. Os animais foram abatidos aos dois meses de idade, sendo previamente submetidos a jejum alimentar de 16 horas. O aleitamento modificou os pesos dos órgãos internos e da gordura omental dos animais, exceto para rins e gordura mesentérica, e não alterou o consumo de alimentos sólidos. A silagem de leite de transição diluída em água comprometeu o desempenho dos bezerros, refletindo no peso dos órgãos internos e de gordura omental, e apresentou pesos absolutos para fígado, baço e gordura omental de 1.072, 186 e 133g, respectivamente. A silagem misturada em leite proporcionou resultados mais semelhantes ao fornecimento exclusivo de leite, demonstrando-se como opção para o aleitamento de bovinos jovens...

This study aimed to evaluate the absolute and relative size of internal organs of Holstein calves which suckled milk, silage transition milk diluted with water or mixed in milk. We used 18 male animals, 60 days old with a mean body weight of 70.3kg (± 11.8kg). The experimental design was completely randomized with three treatments and six replications. The animals were slaughtered at two months, and previously fasted for 16 hours. Breast feeding changed the weights of internal organs and omental fat of animals, except for kidney and mesenteric fat, and did not affect food intake solids. The silage transition milk diluted in water compromised the performance of the calves, reflecting on the weight of internal organs and omental fat, and showed absolute weights for liver, spleen and omental fat of 1072, 186 and 133g, respectively. The silage mixed with milk yielded results more similar to the exclusive milk supply, being an option for suckling calves...

Fungos aeróbios no intestino grosso de borregos e de ovelhas criados em pastagens tropicais / Aerobic fungi in the large intestine of lambs and ewes raised on tropical pastures

Fungi populations were evaluated in large intestine from sheep raised in the North of Minas Gerais, Brazil. The samples were from 39 Santa Inês crossbred ewes and 30 lambs raised on Tanzania grass (Panicum maximum cv. Tanzania), both supplemented with mineral mixtures. The clinical specimens were directly collected from rectal ampoule with sterile swabs. The development of fungal mycelium was observed in all lamb samples and positive cultures were observed in 34 ewe specimens (87.2 percent). After microculture of 40 colonies from lambs, 34 isolates of the genus Aspergillus, three of Paecilomyces spp., one Acremonium sp., and one Trichoderma sp. were identified. Out of the 39 isolates from ewes, 15 Paecilomyces spp., 11 Aspergillus spp., 11 Malbranchea spp., and one Onychocola sp. were identified. The results showed the predominance of the genus Aspergillus in the lambs, while the genus Paecilomyces, Malbranchea, and Aspergillus were predominant, in ewes.

Efeito do extrato aquoso do algodão de seda (Calotropis procera Aiton) sobre a eficiência reprodutiva do carrapato bovino / Effect of aqueous extract of silk cotton (Calotropis procera Ait. R.Br.) on the reproductive performance of Rhipicephalus microplus

O carrapato Rhipicephalus microplus é um dos principais parasitos dos rebanhos bovinos. Diferentes grupos de carrapaticidas sintéticos têm sido empregados para o controle desse ácaro, contudo, o uso constante desses produtos tem selecionado populações resistentes. Na tentativa de minimizar esses impactos tem-se buscado avaliar fitoterápicos para o controle desse parasito. Assim, objetivou-se analisar a eficácia in vitro do extrato aquoso do algodão de seda em diferentes concentrações no controle da postura e eclodibilidade dos ovos do R. microplus. Foram avaliados os efeitos das concentrações 1, 5, 10, 25, 50, 75 e 100% do extrato aquoso obtido juntamente com controles com água destilada ou com cipermetrina, 12,5 mg L-1, associada a organofosforado, 150 mg L-1, sobre a eficiência reprodutiva do carrapato. Foi realizado o biocarrapaticidograma sendo que a eficácia acaricida das concentrações 5, 25 e 100% foram superiores àquela observada para o produto comercial utilizado, que correspondeu a 93,98%, indicando a importância de futuras pesquisas para avaliar o potencial toxigênico e a eficácia in vivo dessa planta para o controle do R. microplus .

The tick Rhipicephalus microplus is one of the main parasites of bovine herds. Different groups of synthetic acaricides have been used to control this mite; however, the constant use of these products has selected resistant populations. In an attempt to minimize these impacts, researchers have tried to evaluate herbal remedies to control this parasite. Therefore, the aim of this study was to analyze the in vitro efficacy of aqueous extract of silk cotton at different concentrations to control the hatchability of R. microplus eggs. The effects of the concentrations 1, 5, 10, 25, 50, 75 and 100% aqueous extract together with controls with distilled water or with cypermethrin, 12.5 mg L-1, associated with organophosphate, 150 mg L-1, were assessed on the reproductive efficiency of the tick. Immersion test was performed and the acaricide efficacy of the 5, 25 and 100% concentrations were higher than that obtained for the commercial product, which corresponded to 93.98%, indicating the importance of further research to assess the toxigenic potential and the in vivo efficacy of this plant to control R. microplus .

Leveduras no rúmen de caprinos e bovinos de corte criados em pastagem tropicais / Yeasts in the rumen of beef steers and goats raised on tropical pasture

Yeasts suplemented in the rumen have been produced benefic interations in the digestion and in the health of the ruminants. This study aimed to quantify, to isolate and, to identify aerobic fungi and yeasts naturally present in the rumen of goats and cattle raised on tropical pastures of the North of Minas Gerais State, Brazil. Samples of 15mL of ruminal juice from 18 hibrid goats and 31 crossbred Nellore steers were used. The physico-chemical characteristics of the samples were evaluated and mycological culture, quantification, and identification of the aerobic fungi were performed. The results indicated the absence of yeasts in the ruminal fluid of steers. However, theses microorganisms were cultured from ruminal juice for all evaluated goats, at an average concentration of 3.2 x 10VCFU/mL. The species Pichia membranifaciens was the most frequently identified yeast, suggesting its participation in the ruminal microbiot of theses small ruminants.

Antimicrobial activity of Lippia sidoides Cham. (Verbenaceae) essential oil against Staphylococcus aureus and Escherichia coli / Atividade antimicrobiana do óleo essencial de Lippia sidoides Cham. (Verbenaceae) contra Staphylococcus aureus e Escherichia coli

The antibacterial effect of Lippia sidoides (rosemary pepper) essential oil was tested against the bacteria Staphylococcus aureus and Escherichia coli isolated from homemade Minas cheese produced in Brazil. The Minimum Inhibitory Concentration (MIC) determined in the Dilution Test was 13 µL oil mL-1 for both bacteria, which characterizes inhibitory action in broth for a 24-hour interaction period. The Minimum Bactericidal Concentration (MBC) determined in the Suspension Test, with one minute of contact, was 25 µL oil mL-1 for both tested bacteria, obtaining at this concentration a bactericidal effect of 99.9 percent on the viable bacterial cells from each sample. Results demonstrated the bacterial activity of Lippia sidoides essential oil against S. aureus and E. coli, suggesting its use as an antibacterial agent in foods.

O efeito antibacteriano do óleo essencial da Lippia sidoides (alecrim-pimenta) foi testado contra as bactérias Staphylococcus aureus e Escherichia coli isoladas de queijo minas artesanal, produzido no Brasil. A Concentração Inibitória Mínima (CIM) determinada por Teste de Diluição foi de 13 µL de óleo mL-1 para as duas bactérias, o que caracteriza ação inibitória em caldo durante vinte quatro horas de interação. A Concentração Bactericida Mínima (CBM) determinada através de Teste de Suspensão, com um minuto de contato, foi de 25 µL de óleo mL-1 para ambas as bactérias testadas, obtendo-se a esta concentração o efeito bactericida de 99,9 por cento sobre as células bacterianas viáveis em cada uma das amostras. Os resultados demonstraram atividade bactericida do óleo essencial de Lippia sidoides contra S. aureus e E. coli, sugerindo a possibilidade do uso como agente antibacteriano em alimentos.

Fungos anaeróbios do rúmen de bovinos e caprinos de corte criados em pastagens tropicais / Ruminal anaerobic fungi of beef cattle and beef goats raised on tropical pastures

The presence of anaerobic fungi structures was evaluated in ruminal juice of beef goats and beef cattle raised in the North of Minas Gerais, Brazil. The strains were collected from 18 Anglo-Nubian crossbred male goats and 23 Nellore crossbred steers during the dry period of the year. Physical-chemical characteristics of the juice were evaluated and direct examination with KOH digestion was performed for anaerobic fungi detection. Structures of these fungi were detected in samples of 14 (77.8 percent) goats and 17 (73.9 percent) steers. The monocentric fungi frequency (56.5 percent) was significantly higher in cattle than polycentric fungi frequency (26.1 percent). This study is the first report of anaerobic ruminal fungi in these ruminants in Brazil and showed high prevalence of theses microorganisms in the ruminal ecosystem of both animals.

C-reactive protein in acute coronary syndrome: association with 3-year outcomes

Inflammatory markers have been associated with clinical outcome in patients with acute coronary syndrome (ACS). The present study evaluated the role of high-sensitivity C-reactive protein (CRP) measurements as a predictor of late cardiovascular outcomes after ACS. One hundred and ninety-nine ACS patients in a Coronary Care Unit from March to November 2002 were included and were reassessed clinically after ~3 years. Clinical variables and CRP levels were evaluated as predictors of major cardiovascular events (MACE, defined as the occurrence of cardiac death, ischemic stroke or myocardial infarction) and mortality. Statistical analyses included Cox multivariable analysis and survival curves (Kaplan-Meier). Of the 199 patients, 11 died within 1 month (5.5 percent). Of the 188 remaining patients, 22 died after a mean follow-up of 2.9 ± 0.5 years. Baseline CRP levels for patients with MACE (N = 57) were significantly higher than those of patients with no events (median = 0.67 mg/L; 25th-75th percentiles = 0.32 and 1.99 mg/L vs median = 0.45 mg/L; 25th-75th percentiles = 0.24 and 0.83 mg/L; P < 0.001). Patients with CRP levels >3 mg/L had a significantly lower survival than the other two groups (1-3 and <1 mg/L; P = 0.001, log-rank test). The odds ratio for MACE was 7.41 (2.03-27.09) for patients with CRP >3 mg/L compared with those with CRP <1 mg/L. For death by any cause, the hazard ratio was 4.58 (1.93-10.86). High CRP levels predicted worse long-term outcomes (MACE and death by any cause) in patients with ACS.

Molecular characterization of Malassezia sympodialis and Malassezia furfur from cattle with and without otitis / Caracterização molecular de isolados de Malassezia sympodialis e Malassezia furfur provenientes de bovinos com e sem otite externa

A molecular study of Malassezia strains isolated from cattle with or without otitis was carried out by random amplified polymorphic DNA analysis (RAPD). DNA was extracted and purified from nine strains of Malassezia sympodialis and fourteen of Malassezia furfur. These microorganisms were collected from eight different bovine herds in Minas Gerais state, Brazil. The RAPD analysis and phenograms did not show the formation of genetically distinct groups among the strain isolated from cattle with or without otitis raised in the same herds. Genetic heterogeneity was observed among Malassezia strains from different geographic origins. These data suggest that genetically similar M. sympodialis and M. furfur strains found as members of the normal ear microbiota could become opportunistically active in the inflammatory process in cattle.

A caracterização molecular de amostras de Malassezia spp., isoladas de bovinos com e sem otite, foi realizada por meio da técnica do DNA polimórfico amplificado ao acaso (RAPD). DNAs de nove amostras de Malassezia sympodialis e quatorze de M. furfur foram extraídos e purificados. Essas amostras foram provenientes de oito diferentes rebanhos bovinos no estado de Minas Gerais, Brasil. A análise de RAPD e os fenogramas não revelaram a formação de grupos geneticamente distintos entre amostras isoladas de bovinos, criados no mesmo rebanho, com ou sem otite. Heterogeneidade genética foi observada entre amostras de diferentes origens geográficas. Os dados sugerem que isolados geneticamente semelhantes e membros da microbiota normal do ouvido podem participar, como oportunistas, no processo inflamatório do conduto auditivo externo de bovinos.