Beef Carcass Contamination by Shiga Toxin-Producing Escherichia coli Strains in an Abattoir in Brazil: Characterization and Resistance to Antimicrobial Drugs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ebselen and cytokine-induced nitric oxide synthase expression in insulin-producing cells

Interleukin-1 (IL-1) may be a mediator of β-cell damage in insulin-dependent diabetes mellitus (IDDM). The IL-1 mechanism of action on insulin-producing cells probably includes activation of the transcription nuclear factor κB (NF-κB), increased transcription of the inducible form of nitric oxide synthase (iNOS) and the subsequent production of nitric oxide (NO). Reactive oxygen intermediates, particularly H2O2, have been proposed as second messengers for NF-κB activation. In the present study, we tested whether ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one), a glutathione peroxidase mimicking compound, could counteract the effects of IL-1β, H2O2 and alloxan in rat pancreatic islets and in the rat insulinoma cell line RINm5F (RIN cells). Some of these experiments were also reproduced in human pancreatic islets. Ebselen (20 μM) prevented the increase in nitrite production by rat islets exposed to IL-1β for 6 hr and induced significant protection against the acute inhibitory effects of alloxan or H2O2 exposure, as judged by the preserved glucose oxidation rates. However, ebselen failed to prevent the increase in nitrite production and the decrease in glucose oxidation and insulin release by rat islets exposed to IL-1β for 24 hr. Ebselen prevented the increase in nitrite production by human islets exposed for 14 hr to a combination of cytokines (IL-1β, tumor necrosis factor-α and interferon-γ). In RIN cells, ebselen counteracted both the expression of iNOS mRNA and the increase in nitrite production induced by 6 hr exposure to IL-β but failed to block IL-1β-induced iNOS expression following 24 hr exposure to the cytokine. Moreover, ebselen did not prevent IL-1β-induced NF-κB activation. As a whole, these data indicate that ebselen partially counteracts cytokine-induced NOS activation in pancreatic β-cells, an effect not associated with inhibition of NF-κB activation.

Reciprocal interference between ants and stingless bees attending the honeydew-producing homopteran Aetalion reticulatum (Homoptera: Aetalionidae)

We evaluated the reciprocal effects between foragers of the ants Camponotus crassus and of the stingless bees Trigona hyalinata on aggregations of the honeydew-producing treehopper Aetalion reticulatum. The interactions were observed in Bauhinia variegata (Caesalpiniaceae) and Mangifera indica (Anacardiaceae) trees. We recorded the presence/absence of each attendant species in homopteran aggregations to test if the observed co-occurrence is lower than that expected by chance. An exclusion experiment was performed in which each attendant species was excluded from aggregations in order to test if an attendant species is more likely to occupy aggregations where the other attendant is not present. We also recorded the number of individuals of each attendant species in homopteran aggregations to search for any correlation between homopteran and attendant abundances. Additionally, we performed experiments using termites (Termitidae, Isoptera) as models to verify if the attendant species have the potential to defend A. reticulatum against natural enemies. The co-occurrence of attendant species was lower than that expected by chance. Homopteran aggregations without stingless bees were more visited by ants than those in which T. hyalinata was present, and vice-versa. The abundance of stingless bees was proportional to homopteran abundance, while ants abundance was not correlated to homopteran abundance. Both attendant species attacked the natural enemies models when we glued the termites ca. 1 cm away from homopteran aggregations, but only ants removed termites glued 5-7 cm away from aggregations. We suggest that the effects of non-formicid attendants should be included as another factor influencing the costs and benefits of ant-homopteran interactions, since honeydew availability for ants also depends on the presence and behavior of interspecific attendants.

Immune Response to the Rhodococcus equi infection in high and low antibody-producing mice (selection IV-A)

Rhodococcus equi is a Gram-positive, facultative intracellular bacterium which infects macrophages and causes rhodococcal pneumonia and enteritis in foals. Recently, this agent has been recognized as an opportunistic pathogen for immunocompromised humans. Several murine experimental models have been used to study R. equi infection. High (H IV-A) and Low (L IV-A) antibody (Ab)-producers mice were obtained by bi-directional genetic selections for their ability to produce antibodies against sheep and human erythrocytes (Selection IV-A). These lines maintain their phenotypes of high and low responders also for other antigens than those of selection (multispeciflc effect). A higher macrophage activity in L IV-A mice has been described for several intracellular infectious agents, which could be responsible for their intense macrophage antigens (Ag)-handling and low Ab production. Due to these differences, L IV-A mice were found to exhibit a better performance to trigger an effective immune response towards intracellular pathogens. The objective of this work was to characterize the immune response of Selection IV-A against R. equi. H IV-A and L IV-A mice were infected with 2.0 × 10 6 CFU of ATCC 33701 +R. equi by intravenous route. With regards to bacterial clearance and survival assays, L IV-A mice were more resistant than H IV-A mice to virulent R. equi. L IV-A mice presented a higher hydrogen peroxide (H 2O 2) and nitric oxide (NO) endogenous production by splenic macrophages than H IV-A mice. L IV-A expressed the most intense cellular response, available by the Delayed-Type Hypersensitivity (DTH) reaction, which activated macrophages and produced more H 2O 2 and NO. The three times higher specific antibodies titres in H IV-A indicated that Selection IV-A maintained the multispecific effect and the polygenic control of humoral and cellular responses also to R. equi.

Hindlimb unloading producing effects on bone biomechanical properties in mature male rats

Aging is associated with decline in muscle mass and strength and reduced bone density. Age-related bone loss is a primary factor in osteoporosis and all individuals are potential candidates for osteoporosis because bone loss with aging occurs in men and women, but less studied in men. To examine the appropriateness of hindlimb elevation, by tail suspension as a model for diminished mechanical loading, and to determine the influence of age on bone responsiveness to skeletal unloading, we use dual X ray absorptiometry (DXA) and digital radiographic images to analyze the response of the femur from mature rats to biomechanical loads. Femurs from male Wistar rats (9-mo-old) were scanned using DXA and DIGORA and measures obtained in ephipyseal and diaphyseal regions of interest. The mechanical testing was divided into compression load to fracture the head and a three-point bending load to fracture the femur middiaphysis. In femoral epiphysis from hindlimb unload (HU), animals presented significant differences between mineral bone content and density assessed by DXA. Detailed regions of femoral epiphysis (head, throcanteric fossa, throcanter and metaphysis) presented significant lower values from radiographic density. Only compressive load necessary to fracture the femoral head neck was also significantly diminished in HU animals. Disuse induced, as in elderly patients, deterioration of the trabecular bone architecture with critical effect on bone fragility. Rats with 21 days of hindlimb unloading can simulate disuse, suggesting that certain sub-regions of their aging bones are more susceptible to fracture, while other, i.e. diaphyses, are not.

Rumen microbial diversity under influence of a polyclonal antibody preparation against lactate-producing and proteolytic bacteria in cows fed different energy sources

Nine ruminally cannulated cows fed different energy sources were used to evaluate an avianderived polyclonal antibody preparation against specific ruminal bacteria and monensin on microbial community diversity. The experimental design was three Latin squares 3 x 3 distinguished by the main energy source in the diet [dry-ground corn grain, high moisture corn silage or citrus pulp]. Inside each Latin square, animals received one of the feed additives per period [control, monensin or polyclonal antibody preparation]. Each period lasted 21 days where 20 were used for treatments adaptation and the last one for sampling collection. Microbial diversity was evaluated by protozoa counts and denaturing gradient gel electrophoresis. Polyclonal antibodies plus citrus pulp (CiPu) addition in the diet resulted in an increase of relative counting of Isotricha protozoa that indicates a possible effect on this ruminal ciliate population. In general lines, in the present experiment, it was not possible to assign that there was a pattern in the structures of amplification of Bacteria and Archaea communities of the ruminal content. Oral passive immunization is a technology that arises as an effective alternative for feed additive production. Further research is still necessary to better understand its mechanisms of action.

Pulmonary and hepatic lesions caused by the dehydropyrrolizidine alkaloid-producing plants Crotalaria juncea and Crotalaria retusa in donkeys

The effects and susceptibility of donkeys to Crotalaria juncea and Crotalaria retusa poisoning were determined at high and low doses. Seeds of C. juncea containing 0.074% of dehydropyrrolizidine alkaloids (DHPAs) (isohemijunceines 0.05%, trichodesmine 0.016%, and junceine 0.008%) were administered to three donkeys at 0.3, 0.6 and 1g/kg body weight (g/kg) daily for 365 days. No clinical signs were observed and, on liver and lung biopsies, the only lesion was a mild liver megalocytosis in the donkeys ingesting 0.6 and 1g/kg/day. Two other donkeys that received daily doses of 3 and 5g seed/kg showed initial respiratory signs 70 and 40 days after the start of the administration, respectively. The donkeys were euthanized following severe respiratory signs and the main lung lesions were proliferation of Clara cells and interstitial fibrosis. Three donkeys ingested seeds of C. retusa containing 5.99% of monocrotaline at daily doses of 0.025, 0.05 and 0.1g/kg for 365 days. No clinical signs were observed and, on liver and lung biopsies, the only lesion was moderate liver megalocytosis in each of the three donkeys. One donkey that received a single dose of 5g/kg of C. retusa seeds and another that received 1g/kg daily for 7 days both showed severe clinical signs and died with diffuse centrilobular liver necrosis. No lung lesions were observed. Another donkey that received a single dose of 2.5g/kg of C. retusa seeds showed no clinical signs. The hepatic and pneumotoxic effects observed are consistent with an etiology involving DHPAs. Furthermore, the occurrence of lung or liver lesions correlates with the type of DHPAs contained in the seeds. Similarly as has been reported for horses, the data herein suggest that in donkeys some DHPAs are metabolized in the liver causing liver disease, whereas others are metabolized in the lung by Clara cells causing lung disease. © 2013 Elsevier Ltd.

Soil and crop residue CO2-C emission under tillage systems in sugarcane-producing areas of southern Brazil

Appropriate management of agricultural crop residues could result in increases on soil organic carbon (SOC) and help to mitigate gas effect. To distinguish the contributions of SOC and sugarcane (Saccharum spp.) residues to the short-term CO2-C loss, we studied the infl uence of several tillage systems: heavy offset disk harrow (HO), chisel plow (CP), rotary tiller (RT), and sugarcane mill tiller (SM) in 2008, and CP, RT, SM, moldboard (MP), and subsoiler (SUB) in 2009, with and without sugarcane residues relative to no-till (NT) in the sugarcane producing region of Brazil. Soil CO2-C emissions were measured daily for two weeks after tillage using portable soil respiration systems. Daily CO2-C emissions declined after tillage regardless of tillage system. In 2008, total CO2-C from SOC and/or residue decomposition was greater for RT and lowest for CP. In 2009, emission was greatest for MP and CP with residues, and smallest for NT. SOC and residue contributed 47% and 41%, respectively, to total CO2-C emissions. Regarding the estimated emissions from sugarcane residue and SOC decomposition within the measurement period, CO2-C factor was similar to sugarcane residue and soil organic carbon decomposition, depending on the tillage system applied. Our approach may define new emission factors that are associated to tillage operations on bare or sugarcane-residue-covered soils to estimate the total carbon loss.

Posibilidades de cooperación y complementación industrial entre América Latina y Japón para la producción y comercialización del hierro y el acero = Possibilities for co-operation and industrial complementation between Latin America and Japan in producing and marketing iron and steel

Incluye Bibliografía

Endophytic fungi producing of esterases: evaluation in vitro of the enzymatic activity using pH indicator

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chemical inhibition of the contaminant Lactobacillus fermentum from distilleries producing fuel bioethanol

The purpose of this study was to determine the Minimum Inhibitory Concentration (MIC) of pure or mixed chemicals for Saccharomyces cerevisiae and Lactobacillus fermentum in the samples isolated from distilleries with serious bacterial contamination problems. The biocides, which showed the best results were: 3,4,4' trichlorocarbanilide (TCC), tested at pH 4.0 (MIC = 3.12 mg/l), TCC with benzethonium chloride (CBe) at pH 6.0 (MIC = 3.12 mg/l) and TCC mixed with benzalkonium chloride (CBa) at pH 6.0 (MIC = 1.53 mg /l). If CBa was used in sugar cane milling in 1:1 ratio with TCC, a 8 times reduction of CBa was possible. This formulation also should be tested in fermentation steps since it was more difficult for the bacterium to develop resistance to biocide. There was no inhibition of S. cerevisiae and there were only antibiotics as an option to bacterial control of fuel ethanol fermentation by S. cerevisiae.

The effect of exogenous fibrolytic enzymes and a ferulic acid esterase-producing inoculant on the fibre degradability, chemical composition and conservation characteristics of alfalfa silage

The objective of this study was to determine the effect of applying fibrolytic enzymes at ensiling, either alone or in combination with a ferulic acid esterase-producing bacterial silage inoculant, on the silage conservation characteristics and nutritive value of alfalfa (Medicago sativa L). Second-cut alfalfa (340 g DM/kg fresh crop) was harvested, wilted, chopped and sub-sampled into 24 batches. Samples were randomly allocated in triplicate to one of four enzyme product treatments supplying endoglucanases and xylanases: none (control), EN1, EN2, EN3; applied alone or in combination with a ferulic acid esterase-producing silage inoculant (FAEI). Treatments were arranged in a 4 x 2 factorial design. All enzyme treatments were applied at 2 ml enzyme product/kg herbage DM, and inoculant was applied at 1 x 10(5) cfu/g fresh herbage. Samples were packed into laboratory-scale silos and stored for 7, 27 or 70 days, and analysed for dry matter (DM) losses, aerobic stability, chemical composition and in vitro ruminal degradability. The use of enzymes did not affect (P>0.05) ensilage DM losses or lactic or acetic acid concentrations after 70 days of ensilage, compared to the control silage. Silage produced using EN1 had lesser neutral detergent fibre (aNDF, P=0.046) and acid detergent fibre (ADF; P=0.006) concentrations than control silage. However, no difference (P>0.05) was observed between the control silage and silage produced with EN1 for aNDF or ADF degradability (NDFD, ADFD). Silages produced with FAEI had greater DM losses (P=0.017) and pH (P<0.001) and lesser NDFD (P=0.019), ADFD (P=0.010) and proportion of lactic acid in the total fermentation products (P=0.006) after 70 days of ensilage, compared to uninoculated silages. The use of fibrolytic enzymes did not have a major effect on the ensilage fermentation of alfalfa, either ensiled alone or with an inoculant. No advantage in ruminal DM or fibre degradability was observed for silages produced with fibrolytic enzymes. The use of a ferulic acid esterase-producing inoculant alone did not improve the nutritive value of alfalfa silage, and did not promote any incremental effects when applied in combination with fibrolytic enzyme products. Crown Copyright (C) 2014 Published by Elsevier B.V. All rights reserved.