Yerba Mate (Ilex paraguariensis) Aqueous Extract Decreases Intestinal SGLT1 Gene Expression but Does Not Affect Other Biochemical Parameters in Alloxan-Diabetic Wistar Rats

Yerba mate (Ilex paraguariensis) is rich in polyphenols, especially chlorogenic acids. Evidence suggests that dietary polyphenols could play a role in glucose absorption and metabolism. The aim of this study was to evaluate the antidiabetic properties of yerba mate extract in alloxan-induced diabetic Wistar rats. Animals (n = 41) were divided in four groups: nondiabetic control (NDC, n = 10), nondiabetic yerba mate (NDY, n = 10), diabetic control (DC, n = 11), and diabetic yerba mate (NDY, n = 10). The intervention consisted in the administration of yerba mate extract in a 1 g extract/kg body weight dose for 28 days; controls received saline solution only. There were no significant differences in serum glucose, insulin, and hepatic glucose-6-phosphatase activity between the groups that ingested yerba mate extract (NDY and DY) and the controls (NDC and DC). However, the intestinal SGLT1 gene expression was significantly lower in animals that received yerba mate both in upper (p = 0.007) and middle (p < 0.001) small intestine. These results indicate that bioactive compounds present in yerba mate might be capable of interfering in glucose absorption, by decreasing SGLT1 expression.

Dynamics of Aeromonas species isolated from wastewater treatment system

Aeromonas are widely distributed in the aquatic environment, and are considered to be emerging organisms that can produce a series of virulence factors. The present study was carried out in a sanitary sewage stabilization pond treatment system, located in Lins, State of Sao Paulo, Brazil. Most probable number was applied for estimation of the genus Aeromonas. Colony isolation was carried out on blood agar ampicillin and confirmed by biochemical characterization. Aeromonas species were isolated in 72.4% of influent samples, and in 55.2 and 48.3% of effluent from anaerobic and facultative lagoons, respectively. Thirteen Aeromonas species were isolated, representing most of the recognized species of these organisms. Even though it was possible to observe a tendency of decrease, total elimination of these organisms from the studied system was not achieved. Understanding of the pathogenic organism`s dynamics in wastewater treatment systems with a reuse potential is especially important because of the risk it represents.

Caesium, potassium and ammonium distributions in different organs of tropical plants

In the present work the distribution of ions in aboveground plant parts was studied in order to establish the suitability of using radiocaesium as a tracer for the plant absorption of nutrients, such as potassium (K(+)) and ammonium (NH(4)(+)). We present the results for the distributions of (137)Cs, (40)K and NH(4)(+) from four tropical plant species: lemon (Citrus aurantifolia), orange (Citrus sinensis), guava (Psidium guajava) and chili pepper (Capsicum frutescens). Activity concentrations of (137)Cs and (40)K were measured by gamma spectrometry and concentrations of free NH(4)(+) ions by a colorimetric method. Similarly to potassium and ammonium, caesium showed a high mobility within the plants, exhibiting the highest values of concentration in the growing parts of the tree (fruits, new leaves, twigs, and barks). A significant correlation between activity concentrations of (137)Cs and (40)K was observed in these tropical plants. The K/Cs discrimination ratios were approximately equal to unity in different compartments of each individual plant, suggesting that caesium could be a good tracer for (40)K in tropical woody fruit species. Despite the similarity observed for the behaviour of caesium and ammonium in the newly grown plant compartments, (137)Cs was not well correlated with NH(4)(+). Significant temporal changes in the NH(4)(+) concentrations were observed during the development of fruits, while the (137)Cs activity concentration alterations were not of great importance, indicating, therefore, that Cs(+) and free NH(4)(+) ions could have distinct concentration ratios for each particular plant organ. (C) 2008 Elsevier B.V. All rights reserved.

Evaluation of Fungal Burden and Aflatoxin Presence in Packed Medicinal Plants Treated by Gamma Radiation

This study was developed to evaluate the fungal burden, toxigenic molds, and mycotoxin contamination and to verify the effects of gamma radiation in four kinds of medicinal plants stored before and after 30 days of irradiation treatment. Eighty samples of medicinal plants (Peumus boldus, Camellia sinensis, Maytenus ilicifolia. and Cassia angustifolia) purchased from drugstores, wholesale, and open-air markets in Sao Paulo city, Brazil, were analyzed. The samples were treated using a (60)Co gamma ray source (Gammacell) with doses of 5 and 10 kGy. Nonirradiated samples were used as controls of fungal isolates. For enumeration of fungi on medicinal plants, serial dilutions of the samples were plated in duplicate onto dichloran 18% glycerol agar. The control samples revealed a high burden of molds, including toxigenic fungi. The process of gamma radiation was effective in reducing the number of CFU per gram in all irradiated samples of medicinal plants after 30 days of storage, using a dose of 10 kGy and maintaining samples in a protective package. No aflatoxins were detected. Gamma radiation treatment can be used as an effective method for preventing fungal deterioration of medicinal plants subject to long-term storage.

Intraspecific variation in 16S rRNA gene of Mycoplasma synoviae determined by DNA sequencing

Mycoplasma synoviae (MS) is an important avian pathogen may cause both respiratory disease and joint inflammation synovitis in poultry, causing economic losses to the Brazilian poultry industry. The genotypic variation in 16S rRNA gene is unknown. Partial sequences of 16S rRNA gene of 19 strains of M. synoviae were sequenced and analyzed in order to obtain molecular characterization and evaluation of the genetic variability of strains from distinct Brazilian areas of poultry production. Different polymorphic patterns were observed. The number of polymorphic alterations in the studied strains ranged from 0 to 6. The nucleotide variations, including deletion, insertion and substitutions, ranged from 3 to 5. The genotypic diversity observed in this study may be explained by spontaneous mutations that may occur when a lineage remains in the same flock for long periods. The culling and reposition in poultry flocks may be responsible for the entry of new strains in different areas. (C) 2008 Elsevier Ltd. All rights reserved.

Intraspecific sequence variation in 16S rRNA gene of Ureaplasma diversum isolates

Ureaplasma diversum infection in bulls may result in seminal vesiculitis, balanoposthitis and alterations in spermatozoids. In cows, it can cause placentitis, fetal alveolitis, abortion and the birth of weak calves. U. diversum ATCC 49782 (serogroups A), ATCC 49783 (serogroup C) and 34 field isolates were used for this study. These microorganisms were submitted to Polymerase Chain Reaction for 16S gene sequence determination using Tact High Fidelity and the products were purified and bi-directionally sequenced. Using the sequence obtained, a fragment containing four hypervariable regions was selected and nucleotide polymorphisms were identified based on their position within the 16S rRNA gene. Forty-four single nucleotide polymorphisms (SNP) were detected. The genotypic variability of the 16S rRNA gene of U. diversum isolates shows that the taxonomy classification of these organisms is likely much more complex than previously described and that 16S rRNA gene sequencing may be used to suggest an epidemiologic pattern of different origin strains. (c) 2011 Elsevier B.V. All rights reserved.

Search for Salmonella spp. in ostrich productive chain of Brazilian southeast region

We analyzed ostriches from an equipped farm located in the Brazilian southeast region for the presence of Salmonella spp. This bacterium was investigated in 80 samples of ostrich droppings, 90 eggs, 30 samples of feed and 30 samples of droppings from rodents. Additionally, at slaughter-house this bacterium was investigated in droppings, caecal content, spleen, liver and carcasses from 90 slaughtered ostriches from the studied farm. Also, blood serum of those animals were harvested and submitted to serum plate agglutination using commercial Salmonella Pullorum antigen. No Salmonella spp. was detected in any eggs, caecal content, liver, spleen, carcass and droppings from ostriches and rodents. However, Salmonella Javiana and Salmonella enterica subsp. enterica 4, 12: i:- were isolated from some samples of feed. The serologic test was negative for all samples. Good sanitary farming management and the application of HACCP principles and GMP during the slaughtering process could explain the absence of Salmonella spp. in the tested samples.

Detection of citrus canker in citrus plants using laser induced fluorescence spectroscopy

Citrus canker is a serious disease caused by Xanthomonas citri subsp. citri bacteria, which infects citrus plants (Citrus spp.) leading to a large economic loss in citrus production worldwide. In Brazil citrus canker control is done by an official eradication campaign, therefore early detection of such disease is important to prevent greater economic losses. However, detection is difficult and so far it has been done by visual inspection of each tree. Suspicious leaves from citrus plants in the field are sent to the laboratory to confirm the infection by laboratory analysis, which is a time consuming. Our goal was to develop a new optical technique to detect and diagnose citrus canker in citrus plants with a portable field spectrometer unit. In this paper, we review two experiments on laser induced fluorescence spectroscopy (LIF) applied to detect citrus canker. We also present new data to show that the length of time a leaf has been detached is an important variable in our studies. Our results show that LIF has the potential to be applied to citrus plants.

Determination of catechins in green tea infusions by reduced flow micellar electrokinetic chromatography

A sulfated-beta-cyclodextrin (s-beta-CD) modified reduced flow micellar electrokinetic chromatography (RF-MEKC) method was developed and validated for the determination of catechins in green tea. The optimal electrolyte consisted of 0.2% triethylamine, 50 mmol/L SDS and 0.8% s-beta-CD (pH = 2.9), allowing baseline separation of five catechins in 4 min. The samples and standards were injected at 0.6 psi for 5 s under constant voltage of -30 kV. Sample preparation simply involved extraction of 2 g of tea with 200 mL water at 95 C under constant stirring for 5 min. The method demonstrated excellent performance, with limits of detection (LOD) and quantification (LOQ) of 0.02-0.1 and 0.1-0.5 mu g/mL, respectively, and recovery percentages of 94-101%. The method was applied to six samples of Brazilian green tea infusions. Epigallocatechin gallate (23.4-112.4 mu g/mL) was the major component, followed by epigallocatechin (18.4-78.9 mu g/mL), epicatechin gallate (5.6-29.6 mu g/mL), epicatechin (4.6-14.5 mu g/mL) and catechin (3.2-8.2 mu g/mL). (C) 2011 Elsevier Ltd. All rights reserved.

Optimization of the rheological properties of probiotic yoghurts supplemented with milk proteins

This study aimed to optimize the rheological properties of probiotic yoghurts supplemented with skimmed milk powder (SMP) whey protein concentrate (WPC) and sodium caseinate (Na-Cn) by using an experimental design type simplex-centroid for mixture modeling It Included seven batches/trials three were supplemented with each type of the dairy protein used three corresponding to the binary mixtures and one to the ternary one in order to increase protein concentration in 1 g 100 g(-1) of final product A control experiment was prepared without supplementing the milk base Processed milk bases were fermented at 42 C until pH 4 5 by using a starter culture blend that consisted of Streptococcus thermophilus Lactobacillus delbrueckii subsp bulgaricus and Bifidobacterium (Humans subsp lactis The kinetics of acidification was followed during the fermentation period as well the physico-chemical analyses enumeration of viable bacteria and theological characteristics of the yoghurts Models were adjusted to the results (kinetic responses counts of viable bacteria and theological parameters) through three regression models (linear quadratic and cubic special) applied to mixtures The results showed that the addition of milk proteins affected slightly acidification profile and counts of S thermophilus and B animal`s subsp lactis but it was significant for L delbrueckii subsp bulgaricus Partially-replacing SMP (45 g/100 g) with WPC or Na-Cn simultaneously enhanced the theological properties of probiotic yoghurts taking into account the kinetics of acidification and enumeration of viable bacteria (C) 2010 Elsevier Ltd All rights reserved

Ageing mallee eucalypt vegetation after fire : insights for successional trajectories in semi-arid mallee ecosystems

A critical requirement in the ecological management of fire is knowledge of the age-class distribution of the vegetation. Such knowledge is important because it underpins the distribution of ecological features important to plants and animals including retreat sites, food sources and foraging microhabitats. However, in many regions, knowledge of the age-class distribution of vegetation is severely constrained by the limited data available on fire history. Much fire-history mapping is restricted to post-1972 fires, following satellite imagery becoming widely available. To investigate fire history in the semi-arid Murray Mallee region in southern Australia, we developed regression models for six species of mallee eucalypt (Eucalyptus oleosa F.Muell. ex. Miq. subsp. oleosa, E. leptophylla F.Muell. ex. Miq., E. dumosa J. Oxley, E. costata subsp. murrayana L. A. S. Johnson & K. D. Hill, E. gracilis F.Muell. and E. socialis F.Muell. ex. Miq.) to quantify the relationship between mean stem diameter and stem age (indicated by fire-year) at sites of known time since fire. We then used these models to predict mean stem age, and thus infer fire-year, for sites where the time since fire was not known. Validation of the models with independent data revealed a highly significant correlation between the actual and predicted time since fire (r = 0.71, P < 0.001, n = 88), confirming the utility of this method for ageing stands of mallee eucalypt vegetation. Validation data suggest the models provide a conservative estimate of the age of a site (i.e. they may under-estimate the minimum age of sites >35 years since fire). Nevertheless, this approach enables examination of post-fire chronosequences in semi-arid mallee ecosystems to be extended from 35 years post-fire to over 100 years. The predicted ages identified for mallee stands imply a need for redefining what is meant by ‘old-growth’ mallee, and challenges current perceptions of an over-abundance of ‘long-unburnt’ mallee vegetation. Given the strong influence of fire on semi-arid mallee vegetation, this approach offers the potential for a better understanding of long-term successional dynamics and the status of biota in an ecosystem that encompasses more than 250 000 km² of southern Australia.

Genome sequence of the clover-nodulating Rhizobium leguminosarum bv. trifolii strain SRDI565

Rhizobium leguminosarum bv. trifolii SRDI565 (syn. N8-J) is an aerobic,motile, Gram-negative, non-spore-forming rod. SRDI565 was isolated from anodule recovered from the roots of the annual clover Trifolium subterraneum subsp. subterraneum grown in thegreenhouse and inoculated with soil collected from New South Wales, Australia. SRDI565has a broad host range for nodulation within the clover genus, however N2-fixationis sub-optimal with some Trifoliumspecies and ineffective with others. Here we describe the features of R. leguminosarum bv. trifolii strain SRDI565, together with genomesequence information and annotation. The 6,905,599 bp high-quality-draft genomeis arranged into 7 scaffolds of 7 contigs, contains 6,750 protein-coding genesand 86 RNA-only encoding genes, and is one of 100 rhizobial genomes sequencedas part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia forBacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

Quick and low cost immobilization of proteinases on polyesters: comparison of lactobacilli cell-envelope proteinase and trypsin for protein degradation

Cell-envelope proteinases (CEPs) are a class of proteolytic enzymes produced by lactic acid bacteria and have several industrially relevant applications. However, soluble CEPs are economically unfavorable for such applications due to their poor stability and lack of reusability. In a quest to prepare stable biocatalysts with improved performance, CEP from Lactobacillus delbrueckii subsp. lactis 313 and trypsin (as a model enzyme) were immobilized onto nonwoven polyester fabrics in a three-step protocol including ethylenediamine activation and glutaraldehyde crosslinking. Immobilization gave protein loading yields of 21.9% (CEP) and 67.7% (trypsin) while residual activity yields were 85.6% (CEP) and 4.1% (trypsin). The activity of the immobilized enzymes was dependent on pH, but was retained at elevated temperatures (40-70°C). An increase in Km values was observed for both enzymes after immobilization. After 70 days of storage, the immobilized CEP retained ca. 62% and 96% of initial activity when the samples were stored in a lyophilized form at -20°C or in a buffer at 4°C, respectively. Both immobilized CEP and trypsin were able to hydrolyze proteins such as casein, skimmed milk proteins and bovine serum albumin. This immobilization protocol can be used to prepare immobilized biocatalyst for various protein degradation processes.

Ocorrência de pinta preta causada por Guignardia citricarpa Kiely em pomares de citros sob manejo orgânico, no município de Montenegro, RS

O cultivo de citros no Brasil tem sido assolado por diversos problemas, como pragas e doenças; entre as últimas, uma das mais importantes e que tem chegado a limitar a cultura, está a pinta preta, causada por Guignardia citricarpa Kiely; que provoca diminuição do tamanho dos frutos, manchas na casca e queda precoce. É de vital importância conhecer a epidemiologia desta doença nas condições locais, para tentativa de controle mais eficiente, principalmente em pomares sob manejo orgânico, nos quais não são utilizados insumos químicos industriais. Com este trabalho, objetivou-se identificar e comparar a incidência e a severidade da doença dentro e entre pomares da bergamoteira ‘Montenegrina’ (Citrus deliciosa Tenore) e tangor ‘Murcott’ (C. reticulata Blanco x C. sinensis Osbeck), a relação entre a severidade e a queda precoce dos frutos; a variação na produção de esporos e a relação de todos estes, com fatores climáticos. Os dados foram coletados entre agosto de 2001 e setembro de 2002, no município de Montenegro, RS, através da contagem direta de todos os frutos das plantas amostradas e, em laboratório, contou-se o número de lesões e seus diâmetros por fruto. Os índices de incidência e severidade da doença foram confrontados com fatores meteorológicos, avaliando-se as implicações no aumento da doença e na queda de frutos. O início dos sintomas deu-se em fevereiro para Murcott e, em março para Montenegrina, agravando-se progressivamente, atingindo valores máximos próximo à colheita. A análise estatística dos dados obtidos mostrou que o número de lesões difere dentro e entre cultivares, mas seus tamanhos parecem ser independentes. A severidade foi relacionada à queda de frutos em Montenegrina, mas não ocorrendo o mesmo em Murcott. Os resultados não evidenciaram influências da umidade relativa do ar, mas temperaturas médias e precipitação pareceram ter relação com o aumento da incidência e da severidade para ambas as cultivares.

Aspectos epidemiológicos da paratuberculose bovina no Rio Grande do Sul

Descrevem-se os achados clínicos e patológicos da paratuberculose em uma criação intensiva de bovinos de leite no municí pio de Capela de Santana, RS. Os sinais clínicos foram observados em oito de um total de 345 bovinos, consistindo em diarréia crônica refratária aos tratamentos, emagrecimento progressivo e queda na produção de leite. As principais lesões macroscópicas, observadas nos oito animais necropsiados, incluíam intestino delgado com acentuado espessamento da parede e superfície mucosa de aspecto reticulado, semelhante às circunvoluções cerebrais, lesão essa perceptível, através da serosa. A luz intestinal estava preenchida com conteúdo fluido e de aspecto leitoso. Os vasos linfáticos do mesentério mostravam-se mais evidentes sendo que alguns tinham aspecto varicoso. Os linfonodos mesentéricos estavam aumentados de volume e, ao corte, fluía grande quantidade de líquido leitoso. Focos de mineralização foram observados na íntima das artérias, nas válvulas cardíacas e na serosa do rúmen. As principais lesões macroscópicas incluíam enterite, linfadenite e linfangite granulomatosa caracterizada por infiltrado inflamatório com macrófagos, células gigantes de Langhans que continham grande quantidade de bacilos álcool-ácido-resistentes. As lesões vasculares consistiam em degeneração e mineralização das túnicas í ntimas e média das artérias de grande calibre associada a proliferação de colágeno. Havia calcificação da serosa do rúmen atrofia hepatocelular difusa e hepatite granulomatosa multifocal. O M. avium subsp. paratuberculosis (Map) foi isolado em amostras de intestino e linfonodos de 8 vacas Holandesas (3,5%) com doença de Johne, dentre 229 amostras cultivadas provenientes de um rebanho leiteiro. Amostras inoculadas em HEYM com micobactina produziram colônias identificadas como Map, segundo as caracterí sticas fenotí picas próprias como: crescimento lento, coloração álcool-ácido-resistente (A.A.R.) e dependência a micobactina. O laboratório de Referência da OIE confirmou a amostra isolada. O M. avium subsp. paratuberculosis (Map) foi isolado em amostras de intestino e linfonodos de 8 vacas Holandesas (3,5%) com doença de Johne, dentre 229 amostras cultivadas provenientes de um rebanho leiteiro Amostras inoculadas em HEYM com micobactina produziram colônias identificadas como Map, segundo as caracterí sticas fenotí picas próprias como: crescimento lento, coloração álcool-ácido-resistente (A.A.R.) e dependência a micobactina. O laboratório de Referência da OIE confirmou a amostra isolada. Não houve isolamento do agente em 221 amostras intestinais quando processadas, após 2 anos de sua colheita. O teste de IDGA aplicado como “screening”, detectou 26 vacas (11,4%) positivas, dentre 228 animais testados e sacrificados em matadouro. O teste de ELISA adsorvido, utilizando o antí geno PPA-3 detectou 125 (39,8%) amostras positivas. O ELISA não adsorvido detectou mais 32 (10,1%) reagentes positivos, dentre os 314 bovinos testados. A prevalência da infecção causada pelo Map em 36 rebanhos leiteiros procedentes de 25 municí pios do Rio Grande do Sul foi estimada em 44,6% das 1316 amostras testadas. A infecção foi identificada em 35 (97,2%) dos 36 rebanhos testados e presentes em todos os municí pios incluí dos. A ocorrência da doença de Johne foi enfatizada, tanto a forma clí nica quanto a infecção subclí nica no Rio Grande do Sul, sugerindo a adoção de medidas de controle sejam aplicadas na proteção dos rebanhos leiteiros nacionais.