Murine colonization model by Campylobacter coli DSPV458.

AIMS: To generate a murine experimental model of colonization by Campylobacter coli DSPV458. METHODS AND RESULTS: Twelve adult Balb/cCmedc female mice were housed in a treated group (T-G) and a control group (C-G) for 4 weeks. Both experimental groups received antibiotics for 5 days during the first week. The T-G was administered with 6.68log10 CFU of C. coli DSPV458 by oesophageal gavage. Necropsies were performed weekly to evaluate translocation and intestinal colonization in the spleen and liver and in the ileum and cecum respectively. Samples were cultured to quantify intestinal microbiota members. Faeces were cultured weekly for a C. coli DSPV458 count. Campylobacter coli DSPV458 was isolated from all the inoculated mice. The recovered level of C. coli DSPV458 was, on average, 6.9 log10 CFUg-1 , 8.0 log10 CFUg-1 and 1.6 log10 CFUg-1 in faeces, cecum and ileum respectively. Colonization by C. coli DSPV458 does not alter the normal clinical and physiological status. CONCLUSIONS: Campylobacter coli DSPV458 does not have an invasive capacity, and the model is suitable for evaluating strategies to reduce intestinal loads. SIGNIFICANCE AND IMPACT OF STUDY: Farm animals have an important impact on thermotolerant Campylobacter transmission to humans. Extremely few colonization models by C. coli have been reported to date. In food-producing animals, infection is mild or absent and thermotolerant Campylobacter colonize the intestines of animals. Colonization models are specific models that do not cause infection as they do not generally result in diarrhoea or other signs of disease. Therefore, this model will allow to evaluate the evolution of colonization by thermotolerant Campylobacter and the alternative tools development to antibiotics that limit their colonization in food-producing animals.

In vitro screening and in vivo colonization pilot model of Lactobacillus plantarum LP5 and Campylobacter coli DSPV 458 in mice.

The objective of this work was to determine the antibacterial effect of Lactobacillus plantarum strains of pork origin against Campylobacter coli strains, and to conduct experimental colonization pilot models in mice for both microorganisms. Inhibition assays allowed evaluation and selection of L. plantarum LP5 as the strain with the highest antagonistic activity against C. coli and with the best potential to be used in in vivo study. Adult 6-week-old female Balb/cCmedc mice were lodged in two groups. The treated group was administered with 9.4 log10CFU/2 times/wk of L. plantarum LP5. L. plantarum LP5 was recovered from the feces and cecum of the inoculated mice. However, when bacteria stopped being administered, probiotic counts decreased. Experimental colonization with C. coli was carried out in five groups of mice. All animals were treated with antibiotics in their drinking water to weaken the indigenous microbiota and to allow colonization of C. coli. Four groups were administered once with different C. coli strains (DSPV458: 8.49 log10CFU; DSPV567: 8.09 log10CFU; DSPV570: 8.46 log10CFU; DSPV541: 8.86 log10CFU, respectively). After 8 h, mice inoculated with different C. coli strains were colonized because the pathogen was detected in their feces. L. plantarum LP5 tolerated the gastrointestinal conditions of murine model without generating adverse effects on the animals. C. coli DSPV458 colonized the mice without causing infection by lodging in their digestive tract, thus generating a reproducible colonization model. Both models combined could be used as protection murine models against pathogens to test alternative control tools to antibiotics.

Yeast derivatives and wheat germ in the adult diet modulates fecundity in a tephritid pest.

Anastrepha fraterculus (Wiedemann), a pest of great economic importance in South America, needs urgently to be controlled by environmentally friendly methods such as the sterile insect technique for which mass rearing of insects is required. Because oogenesis takes place during the adult stage, mass-rearing facilities should provide the females a diet that maximizes egg production at the lowest cost. Accordingly, we investigated the effect of artificial protein sources in the adult diet (yeast derivatives of different cost but with similar amino acids profiles, and the addition of wheat germ) on fecundity. Additionally, we evaluated different ratios of yeast derivatives or wheat germ on ovary maturation, fecundity, and fertility as well as their association with the nutrient content of females. Females fed hydrolyzed yeast and yeast extract attained the highest fecundity level, and those fed brewer's yeast the lowest. Reducing the amount of hydrolyzed yeast, an expensive protein source, in the diet negatively affected fecundity and ovary maturation. Increasing the amount of brewer's yeast, a low-cost protein source, did not favor fecundity. The addition of wheat germ in the adult diet improved fecundity regardless of the yeast derivate considered. Percentage of egg hatch was not affected by the diet. Nutrient content of A. fraterculus females varied according to the adult diet provided and mating status. Our findings provide novel baseline information to understand the role of nutrition on reproductive performance of A. fraterculus females and are discussed in the context of resource allocation. They also provide valuable advances in the search for cost-effective adult diets at fruit fly mass rearing facilities.

Predictors of Disordered Eating in Young Males.

Recent findings suggest that disordered eating (DE) symptomatology may be underestimated in the male population. The present study examined depressive symptomatology as a potential mediator of the relationships between body image dissatisfaction, strategies to change body weight and muscles, media pressure, and DE (emotional, restrained and emotional eating) in 260 male undergraduates who completed a self-reported questionnaire. Path analyses indicated that media influence and strategies to decrease body weight had direct positive effects on depressive symptomatology, which in turn predicted emotional eating. Media influence had a direct positive effect on emotional eating, whereas strategies to decrease body weight did not exhibit a direct effect on emotional eating. Therefore, the latter pathway was removed from the model. The link between media pressure, strategies to decrease body weight and emotional eating was partially mediated by depressive symptomatology. The present findings can inform the development and implementation of prevention and education programs for DE in schools and universities.

A youth mentor-led nutritional intervention in urban recreation centers: a promising strategy for childhood obesity prevention in low-income neighborhoods.

B'More Healthy Community for Kids (BHCK) is an ongoing multi-level intervention to prevent childhood obesity in African-American low-income neighborhoods in Baltimore city, MD. Although previous nutrition interventions involving peer mentoring of youth have been successful, there is a lack of studies evaluating the influence of cross-age peers within interventions targeting youth. This article evaluates the implementation of the BHCK intervention in recreation centers, and describes lessons learned. Sixteen youth leaders delivered bi-weekly, interactive sessions to 10- to 14-y olds. Dose, fidelity and reach are assessed, as is qualitative information regarding what worked well during sessions. Dose is operationalized as the number of interactive sessions, and taste tests, giveaways and handouts per session; fidelity as the number of youth leaders participating in the entire intervention and per session and reach as the number of interactions with the target population. Based on a priori set values, number of interactive sessions was high, and number of taste tests, giveaways and handouts was moderate to high (dose). The number of participating youth leaders was also high (fidelity). Of the 14 planned sessions, the intervention was implemented with high/moderate reach. Data suggest that working with cross-age peers is a promising nutritional intervention for recreation centers.

Chlorpyrifos induces cytotoxicity via oxidative stress and mitochondrial dysfunction in HepG2 cells.

Chlorpyrifos (CPF), a widely used broad-spectrum organophosphate pesticide, has been associated with various adverse health effects in animals and humans. While its primary mechanism of action involves the irreversible inhibition of acetylcholinesterase, secondary mechanisms have also been suggested. The aim of the present study was to explore the secondary mechanisms of action involved in CPF-induced acute cytotoxicity using human hepatocarcinoma HepG2 cells. In particular, we investigated oxidative stress and mitochondrial function by assessing reactive oxygen species (ROS) generation, lipid peroxidation (LPO) and mitochondrial membrane potential (ΔΨm) alteration. Results showed that 24-h exposure to CPF (78.125-2500 µM) decreased cell viability in a concentration-dependent manner (IC50 = 280.87 ± 26.63 µM). Sub-toxic CPF concentrations (17.5, 35 and 70 µM) induced increases in ROS generation (by 83%), mitochondrial superoxide (by 7.1%), LPO (by 11%), and decreased ΔΨm (by 20%). CPF also upregulated Nrf2 protein expression, indicating the role of the latter in modulating the cellular response to oxidative insults. Overall, our findings suggest that CPF caused hepatotoxicity through oxidative stress and mitochondrial dysfunction. Given the re-emerging use of CPF, this study emphasizes the need for comprehensive analysis to elucidate its toxicity on non-target organs and associated mechanisms.

Effect of polyphenols on enniatins-induced cytotoxic effects in mammalian cells.

Enniatins (ENs) are fungal secondary metabolites produced by genus Fusarium. The ENs exert antimicrobial and insecticidal effect, and has also been demonstrated cytotoxic effects on several mammalian cell lines. On the other hands, it has been proved that natural polyphenols have antioxidant effect. In this study, cell effects at low levels of exposure of four ENs (A, A(1), B and B(1)) and five polyphenols (quercetin, quercetin-3-ß-D-glucoside, rutin, myricetin and t-pterostilbene) present in wine; and the cytoprotective effect of these polyphenols exposed simultaneously with ENs in Chinese Hamster Ovary (CHO-K1) cells, were studied. Cell effects were determined by the MTT test after 24 h of exposure. All ENs showed cytotoxic effect. The IC(50) obtained ranged from 4.5 ± 1.2 to 11.0 ± 2.7 µM. The concentration of polyphenols tested ranged from 5 to 50 µM. Polyphenols did not show cytotoxicity and the cytoprotective effect of polyphenols varies depending on the EN tested. The cytoprotective effect of polyphenols in CHO-K1 cells exposed to ENs was as follow: quercetin, from 24 to 84%; quercetin-3-ß-D-glucoside, from 12 to 76%; rutin, from 17 to 83%; myricetin, from 16 to 92% and pterostilbene from 25 to 100%. All polyphenols protected CHO-K1 cells against EN A(1) exposure.

Multi-mycotoxin occurrence in feed, metabolism and carry-over to animal-derived food products: A review.

The world requests for raw materials used in animal feed has been steadily rising in the last years driven by higher demands for livestock production. Mycotoxins are frequent toxic metabolites present in these raw materials. The exposure of farm animals to mycotoxins could result in undesirable residues in animal-derived food products. Thus, the potential ingestion of edible animal products (milk, meat and fish) contaminated with mycotoxins constitutes a public health concern, since they enter the food chain and may cause adverse effects upon human health. The present review summarizes the state-of-the-art on the occurrence of mycotoxins in feed, their metabolism and carry-over into animal source foodstuffs, focusing particularly on the last decade. Maximum levels (MLs) for various mycotoxins have been established for a number of raw feed materials and animal food products. Such values are sometimes exceeded, however. Aflatoxins (AFs), fumonisins (FBs), ochratoxin A (OTA), trichothecenes (TCs) and zearalenone (ZEN) are the most prevalent mycotoxins in animal feed, with aflatoxin M1 (AFM1) predominating in milk and dairy products, and OTA in meat by-products. The co-occurrence of mycotoxins in feed raw materials tends to be the rule rather than the exception, and the carry-over of mycotoxins from feed to animal source foods is more than proven.

Protective effect of Lactiplantibacillus plantarum LP5 in a murine model of colonisation by Campylobacter coli DSPV458.

Thermotolerant Campylobacter species are the leading cause of foodborne bacterial diarrheal disease worldwide. Campylobacter coli, abundant in pigs and pork products, have been identified as a source of human infection. In this study, we propose the use of Lactiplantibacillus plantarum LP5 as a probiotic to reduce colonisation of this intestinal pathogen in a murine colonisation model of C. coli DSPV458. Six-week-old adult female Balb/cCmedc mice were housed in groups: Control, Campy and Pro-Campy. Control and Pro-Campy groups received antibiotics for 5 days and the Campy group for 12 days. Pro-Campy group was inoculated for 7 days with 8.78 log10 cfu total of L. plantarum LP5 suspended in De Man, Rogosa and Sharpe broth. All groups were inoculated with 6.72 log10 cfu of C. coli DSPV458 suspended in brain heart infusion broth. L. plantarum LP5 was recovered only in the Pro- Campy group. C. coli DSPV458 was recovered at higher levels in the Control and Campy groups. The differences with the Pro-Campy group were significant. As regards faeces, Control and Campy groups reached 7.41 and 7.84 log10 cfu/g, respectively, and the Pro-Campy group only 4.62 log10 cfu/g. In the caecum, Control and Campy groups reached 8.01 and 9.26 log10cfu/g, respectively, and the Pro-Campy group only 4.51 log10 cfu/g. In the ileum, Control and Campy groups reached 3.43 and 3.26 log10 cfu/g, respectively, and the Pro-Campy group did not show detectable levels. The reduction of C. coli DSPV458 in the Pro-Campy group compared to the Control group in faeces, caecum and ileum was 99.55, 99.98 and 100%, respectively. Animals were maintained under normal health conditions, and haematological parameters were within the standard values for Balb/cCmedc. The incorporation of a probiotic generated a protective effect in the mice colonisation model. The protective effect would also apply to intestinal colonisation by indigenous enterobacteria. Therefore, the strategy used in this study is of great importance to understand the protection mechanisms in a murine model, as well as its application in food-producing animals.

Bioinformatic analysis of post-transmission viral readaptation in Argentine patients with acute HIV-1 infection.

During the acute phase of HIV-1 infection, a strong readaptation occurs in the viral population. Our objective was to analyze the post-transmission mutations associated with escape to the cytotoxic immune response and its relationship with the progression of the infection. In this study, a total of 17 patients were enrolled during acute/early primary HIV infection and 8 subjects that were the HIV positive partner resulting in 8 transmission pairs. Genotyping of the genetic polymorphisms of HLA class I A and B was performed using PCR-SSOP. Viral RNA extraction was from plasma. 570 single Gag-gene amplifications were obtained by limiting-dilution RT-PCR. Epitope prediction was performed with NetMHC CBS prediction server for the 19 HLA-A and B alleles. Cytotoxic response prediction was performed by using the IEDB Analysis Resource. From our results, we deduce that the transmitted CTL / gag escape frequency in the founder virus was at least double compared to the post-transmission events. Additionally, by means of an algorithm that combines these frequencies, we observed that the founder viruses better adapted to the HLA A / B alleles of the recipient could contribute to a greater progression of the infection. Our results suggest that there is a large adaptation of HIV-1 to the HLA A / B alleles prevalent in our population. However, despite this adaptive advantage, the virus needs to make "readjustments" through new escape and compensatory mutations. Interestingly, according to our results, this readaptation could have a role in the progression of the infection.

Pesticide residue determination in surface waters by stir bar sorptive extraction and liquid chromatography/tandem mass spectrometry.

In this stir bar sorptive extraction (SBSE) method, 16 pesticides were extracted from surface water samples by sorption onto 1 mm polydimethylsiloxane layer coated on a 10-mm-length stir bar magnet. After liquid desorption of the analytes with 1 ml of methanol, the detection was performed on a liquid chromatography-tandem mass spectrometry with a triple quadrupole (QqQ) analyzer using selected reaction monitoring mode via electrospray ionization. Parameters affecting SBSE operation, including sample volume, salt addition, extraction time, stirring rate, and desorption conditions, have been evaluated. The optimized SBSE method required two 50 ml aliquots of surface water samples, one aliquot was added of 30% NaCl and stirred at 900 rpm during 1 h for testing five pesticides with log K(o/w) < 3, and the other aliquot was directly extracted following the same procedure for the rest of the pesticides with log K(o/w) > 3. The method was validated in spiked surface water samples at limits of quantifications (LOQs) and ten times the LOQs showing recoveries <62%, and the LOQs reached were from 0.03 microg l(-1) for diazinon to 3 microg l(-1) for simazine. The proposed methodology was applied to the determination of these compounds in samples from Albufera Lake and surrounding channels, showing that SBSE is a powerful tool for routine control analysis of pesticide residues in surface water.

Beauvericin and enniatin B effects on a human lymphoblastoid Jurkat T-cell model.

Several mycotoxins exert their effect on the immunological system; some are classified as immunotoxic. Jurkat T-cells were used to study toxic effects of beauvericin (BEA) and enniatin B (ENN B). Both are not legislated mycotoxins with increasing presence in feed and food. Concentrations studied were from 1 to 15 µM at 24, 48 and 72 h. Cell death by increasing the percentage of apoptotic/necrotic cells was: BEA > ENN B. IC50 values ranged from 3 to 7.5 µM for BEA. ENN B 15 µM decreased viability (21-29%). The percentage of apoptotic/necrotic cells was BEA > ENN B at 24 h but not at 48 h. Caspase-3&7 activation profile varied, although both mycotoxins increased this activation. No difference in ROS production for any mycotoxin was observed. Arrest in S phase for both mycotoxins was obtained. BEA increased the percentage of DNA in the tail (18% and 20%) with respect to the control, whereas not for ENN B. In summary, cytotoxicity of BEA involved mitochondrial alterations; while ENN B only at highest concentrations and time assayed. BEA had cell cycle disturbances and apoptotic and apoptotic/necrotic cells increased; for ENN B these were not evident. Different toxic responses in Jurkat T-cells may be involved in BEA and ENN B toxicity.

Reaction of zearalenone and α-zearalenol with allyl isothiocyanate, characterization of reaction products, their bioaccessibility and bioavailability in vitro.

This study investigates the reduction of zearalenone (ZEA) and α-zearalenol (α-ZOL) on a solution model using allyl isothiocyanate (AITC) and also determines the bioaccessibility and bioavailability of the reaction products isolated and identified by MS-LIT. Mycotoxin reductions were dose-dependent, and ZEA levels decreased more than α-ZOL, ranging from 0.2 to 96.9% and 0 to 89.5% respectively, with no difference (p⩽0.05) between pH 4 and 7. Overall, simulated gastric bioaccessibility was higher than duodenal bioaccessibility for both mycotoxins and mycotoxin-AITC conjugates, with duodenal fractions representing ⩾63.5% of the original concentration. Simulated bioavailability of reaction products (α-ZOL/ZEA-AITC) were lower than 42.13%, but significantly higher than the original mycotoxins. The cytotoxicity of α-ZOL and ZEA in Caco-2/TC7 cells was also evaluated, with toxic effects observed at higher levels than 75µM. Further studies should be performed to evaluate the toxicity and estrogenic effect of α-ZOL/ZEA-AITC.

Cytoprotective effect of resveratrol diastereomers in CHO-K1 cells exposed to beauvericin.

Beauvericin (BEA) causes cytotoxicity, lipid peroxidation and reactive oxygen species in CHO-K1 cells. Resveratrol (RSV) is a polyphenol with multiple biological properties, including antioxidant effects. RSV has two forms: trans and cis. The aims of this study were to determine the cytoprotective effect of trans-RSV and diastereomers mixtures (50:50 trans/cis-RSV and 70:30 trans/cis-RSV) incubated alone and in combination with BEA in ovarian (CHO-K1) cells. The results demonstrated that cell viability increases (from 9% to 77%) when they were exposed to low concentration of RSV. Moreover, when the cells were pre-treated with RSV and then exposed to BEA, a cytoprotective effect (from 25% to 76%) and a ROS production diminution (from 27% to 92%) were observed, with respect to cells exposed to BEA without previous RSV exposure. RSV pre-treatment decreased the MDA levels (from 15% to 37%) when it is compared with cells exposed only to BEA. Therefore, it can be concluded that RSV could reduce the toxicological risk produced by BEA when they are in combination.

Alternariol-induced cytotoxicity in Caco-2 cells. Protective effect of the phenolic fraction from virgin olive oil.

The extra virgin olive oil (EVOO) has been associated to antioxidant effects. The mycotoxin alternariol (AOH) can contaminate olives. The aims of this work were to determine the cytotoxic effects and reactive oxygen species (ROS) produced by AOH, tyrosol and oleuropein (two polyphenols of olive oil) and a real EVOO extract in Caco-2 cells. The MTT assay and the ROS production by the H2-DCFDA probe were used. Results demonstrated that AOH reduces cellular proliferation depending on concentration, whereas tyrosol and oleuropein did not (12.5-100 µM). The combination of AOH + oleuropein (50 µM) increased cell proliferation (24%) whereas, AOH + tyrosol decreased (47%) it. Besides, AOH increased ROS generation depending on time and concentration. Oleuropein + AOH decreased ROS production. However, 25 µM of tyrosol increased 1.2-fold the ROS production. Respect to the EVOO extract, cytoprotective effect (151%) was evidenced, even with the combination EVOO extract + AOH (15%-55% respect to cells exposed to AOH alone). ROS generation was significantly reduced compared to ROS generation produced by 25 µM of AOH alone. The phenolic antioxidant of EVOO decreases cytotoxicity and ROS production in Caco-2 cells exposed to AOH. Thus, polyphenols of EVOO could contribute to diminish the toxicological risk that mycotoxins can produce to humans.

Serological response (Western blot) to fractions of Mycobacterium tuberculosis sonicate antigen in tuberculosis patients and contacts.

OBJECTIVE: To assess the serological response to fractions of Mycobacterium tuberculosis sonicate antigen by Western blot analysis in patients with tuberculosis and contacts. METHODS: We studied 71 individuals including 43 patients with active tuberculosis, 16 contacts and 12 healthy blood donors. For Western blot analysis, M. tuberculosis (H37Rv strain) sonicate antigen extract was fractionated by electrophoresis on polyacrylamide gel (SDS-PAGE). RESULTS: We obtained antibody responses directed against four antigenic fractions with molecular weights of 71, 65, 26-38 and 19 kDa. Sixty per cent of pleural tuberculosis and 52.4% of smear-positive pulmonary tuberculosis had whole responses against all four fractions; there were no partial responses in these groups. For patients with smear-negative pulmonary tuberculosis whole responses were 17.6% and partial responses 41.2%. All contacts whose tuberculin tests converted from negative to positive (three cases) reacted exclusively against the 19 kDa fraction. CONCLUSIONS: Western blot-positive results in patients with pleural and smear-positive pulmonary tuberculosis were characterised by a whole pattern against all four antigenic fractions, whereas patients with smear-negative pulmonary tuberculosis showed heterogeneous results. The exclusive response against the 19 kDa fraction observed in contacts with tuberculin conversion could help to identify candidates for preventive therapy.

Genotoxicity of six pesticides by Salmonella mutagenicity test and SOS chromotest.

Two in vitro tests (Ames test and SOS chromotest), one for bacterial mutagenicity and one for primary DNA damage, were assayed to determine the genotoxic activity of 6 pesticides (atrazine, captafol, Captan, chlorpyrifosmethyl, molinate and tetrachlorvinphos). Assays were carried out both in the absence and presence of S9 fractions of liver homogenate from rat (Sprague-Dawley) pretreated with Aroclor 1254. Captan and captafol were genotoxic on both the Ames test and the SOS chromotest. Comparisons with mutagenesis data in Salmonella indicated that the SOS assay detected as genotoxic the pesticides that were mutagenic on the Salmonella test. Non-genotoxic effects were not detected in vitro either in the Salmonella/microsome assay nor in the SOS chromotest when bacterial tester strains were exposed to atrazine, molinate, chlorpyrifosmethyl and tetrachlorvinphos in the absence or presence of S9 mix.

Radiation doses to paediatric patients undergoing micturating cystourethrography examinations and potential reduction by radiation protection optimization.

Dose-area product values were measured in paediatric micturating cystourethrography examinations in two dedicated facilities. The results are compared, taking into account features of the X-ray rooms and differences in examination protocol. Several optimization procedures were simulated or implemented, in order to estimate the dose savings which could be attained. Potential savings identified in the dose-area values reach about 85% for both centres.

Measurement of radiation doses in the most frequent simple examinations in paediatric radiology and its dependence on patient age.

Radiation doses to patients were measured in four X-ray rooms specifically devoted to paediatric radiology, from two hospitals. The study was performed for the most frequent simple examinations, namely abdomen, hip and pelvis, skull, spine and chest. Patients were classed into four different age groups: 0.1-1 year, greater than 1-5 years, greater than 5-10 years and greater than 10-14 years. Operating X-ray generator parameters and entrance surface doses were recorded for all groups. Representative values were obtained for standard working conditions prior to any correcting action being taken. Dose values are reported, and some of the differences between the results found in the rooms for each examination are discussed. Without attempting to relate adult and paediatric radiology, the entrance surface doses measured and the provisionally recommended CEC values for similar examinations in adult patients are compared.

Dose-area product values in frequently performed complex paediatric radiology examinations.

Mean values of the dose-area product for barium meal, micturating cystourethrography and intravenous urography examinations from three health centres and for four age groups are presented in this study. Values measured are between 13% and 86% higher than results previously reported, which justifies the importance of applying procedures which reduce risks to patients.