Desiccated Liver

Polyphenols in red wine are bioactive compounds with positive effects on health and disease prevention. White grape musts and wines have a lower concentration of phenolic compounds compared to the red ones and are therefore considered less beneficial to health. In Andalucía, a region located in the South West of Spain, Pedro Ximenez white grapes are desiccated under the sun for a week before they are pressed and the juice (must) is obtained. This ancient procedure increases the variety and content of polyphenols present in the Pedro Ximenez must (PXM). We incorporated PXM into the daily diet of aged Mus spretus mice (24 months) and investigated their properties by comparing several parameters determined in these old mice with those measured in young mice (two months old). Biochemical, histological, and transcriptional analyses indicated that PXM exhibited potent antioxidant properties, promoted the normalization of the biotransforming ability of several cytochromes, i.e., the P450 family, in the liver, and regularized hepatic apoptosis, promoting proliferation instead. Our data indicated that PXM possesses a profound ability to promote liver regeneration in terms of both the structure and the function, thus contributing to a healthy aging process.

Enterococci are included in the United States National Antimicrobial Resistance Monitoring System to track antibiotic resistance among commensal Gram-positive enteric bacteria, largely due to their high abundance in food animals and in retail meat. In the U.S. cattle industry, macrolides are used to prevent and control liver abscesses, which cause significant economic losses. Previous studies have suggested that feeding tylosin and the intensity of the pen environment, both expand and sustain respectively the prevalence of multidrug resistance among enterococci in feedlot cattle. This has led to research into alternative feed supplements and improved stewardship practices. In a randomized controlled trial, we measured the impact of a probiotic and an altered pen environment on antimicrobial resistance among faecal Enterococcus spp. in cattle fed tylosin. Supplementing cattle with an Enterococcus faecium and Saccharomyces cerevisiae-based probiotic yielded the isolation of E. faecium of the probiotic sequence type (ST296) from faecal and environmental samples in treatment groups, as well as from cattle and the manure pack in nearby pens. Of importance, the probiotic strain also was found in a desiccated and milled manure pack sample taken 120 days after the initial trial ended. Phylogenetic and SNP analyses revealed clonal survival and spread compatible with faecal-environmental-oral recycling of the probiotic strain within and among cattle and pens. The increase in prevalence of the ST296 strain occurred concomitant with a decrease in ST240, the dominant sequence type associated with ermB and tet(M) resistance genes in this trial. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrate that a macrolide-susceptible probiotic Enterococcus faecium ST296 strain fed to beef cattle becomes fully embedded in the microbial community cycling of bacteria via faecal-environmental-oral transmission within and among feedlot pens. An initial investment in feeding the probiotic is thereby leveraged into expanding numbers of susceptible bacteria in cattle and their environment, even among those cattle fed tylosin. © 2019 The Society for Applied Microbiology.

Objective: To investigate the impact of ultrasonic assisted rapid processing technique combined with the environment friendly reagent (which can be utilized in fixing,dehydrating and clearing) on processing tumor biopsy specimens and the subsequent target detection. Methods: Postoperative tissue samples of 56 cases of breast cancer, colorectal cancer, lung cancer, stomach cancer, liver mass, bladder mass, uterus mass were obtained at the National Cancer Center, Cancer Hospital, Chinese Academy of Medical Sciences from February to April, 2017. Three specimens ranging in size from 1 to 3 mm were collected from each sample, and were separated into control group (traditional tissue-processing method); experiment group 1 (3.7% neutral buffered formaldehyde fixation, composite environment friendly reagent and ultrasonic assisted rapid processing) and experimental group 2 (composite environment friendly reagent direct fixation, higher temperature and longer time for tissue processing). Two pathologists blinded to the experimental groups scored totally the nuclear, cytoplasmic, and membrane staining of 43 cases of immunohistochemistry (IHC), four HER2 fluorescence in situ hybridization (FISH), 20 extracted DNA quality and four EGFR gene mutation detection in lung adenocarcinoma; the results were compared with the control group. Results: There was no difference in the IHC staining, HER2 FISH, the DNA quality, and EGFR genetic results between experimental group 1 and control group. For experiment group 2, comparing results of IHC staining, HER2 FISH and the quality of DNA, there was no obvious difference from control group and experiment group 1, but might show an increase in the background of IHC staining. The difference between the treatment temperature and time in the experimental group 2 did not affect the results of the gene mutation detection. Conclusions: Environment freindly reagent and ultrasonic assisted rapid processing equipment could be used for rapid processing and diagnosis for tumor biopsies. Using complex environment-friendly reagents supplement fixation, higher treatment temperature and longer treatment time do not significantly affect the IHC, FISH and

Lemon (Citrus limon) is the third most important species of citrus in the world, while Spain is the major producer in Europe. Numerous beneficial effects of lemon are known, which explains their use in traditional medicine. The paper describes the effect of dietary dehydrated lemon peel (a sub-product of the lemon industry) on the growth, immune and antioxidant status of gilthead seabream (Sparus aurata L.) over a period of 30 days. Fish fed diets enriched with dehydrated lemon peel (1.5% and 3%) for 15 days showed improved growth and both humoral (seric immunoglobulin M) and cellular (peroxidase activity and phagocytic ability of head kidney leucocytes) immunity, as well as the expression of some immune-related genes (nkefa, il1β, igth and csfr1). However, decreases growth promotion was observed after thirty days of trial. Neither the anti-oxidant enzymes activity nor the expression of several anti-oxidants and anti-stress genes in liver was improved by the diet. The possible inclusion of dehydrated lemon peel in fish diets for its immunostimulant effects is discussed.

The effect of a product made of dehydrated apples enriched with mandarin juice by vacuum impregnation on markers of oxidative stress (plasma antioxidant capacity, carbonyl groups (CGs), 8-hydroxydeoxyguanosine (8OHdG) and α-tocopherol) was tested in rats. Six groups of animals were studied: one group was fed a standard diet; two groups were supplemented with dehydrated apple either impregnated or not with mandarin juice throughout 28 days; and three groups (one unsupplemented and two supplemented) were additionally treated with tamoxifen (TAM) for 21 days used for induction of oxidative stress. The rats treated with TAM showed an increase in aminotransferases, CGs and 8OHdG. All of these effects were significantly decreased in the animals after apple snack consumption; the addition of mandarin juice into the apple mainly accounts for increased levels of α-tocopherol in plasma and liver. These findings suggest that the food product have a protective action against oxidative stress induced by TAM in rats.

Salmonella enterica serovar Enteritidis strain E40 filaments were developed under conditions of a reduced water activity (a(w)) of 0.95 in tryptic soy broth (TSB) or tryptic soy agar (TSA) supplemented with 8% or 7% NaCl, respectively. Filament formation was accompanied by an increase of biomass without an increase in CFU and was affected by incubation temperature and the physical milieu. The greatest amount of filaments was recovered from TSA with 7% NaCl and incubation at 30°C. Within 2 h of transfer to fresh TSB, filaments started to septate into normal-sized cells, resulting in a rapid increase in CFU. S. Enteritidis E40 filaments were not more tolerant of low- or high-temperature stresses than nonfilamented control cells. However, there was greater survival of filaments in 10% bile salts after 24 to 48 h of incubation, during pH 2.0 acid challenge for 10 min, and under desiccation on stainless steel surfaces at 25°C and 75.5% relative humidity for 7 days. S. Enteritidis E40 filaments invaded and multiplied within Caco-2 human intestinal epithelial cells to a similar degree as control cells when a comparable CFU of filaments and control cells was used. S. Enteritidis E40 filaments established a successful infection in mice via intragastric inoculation. The filaments colonized the gastrointestinal tract and disseminated to the spleen and liver at levels comparable to those attained by control cells, even when animals were inoculated with 10- to 100-fold fewer CFU. To our knowledge this is the first demonstration of virulence of stress-induced Salmonella filaments in vitro and in vivo. Formation of filaments by Salmonella in food products and food processing environments is significant to food safety, because detection and quantitation of the pathogen may be compromised. The finding that these filaments are virulent further enhances their potential public health impact.

Using HPLC-ESI-MS, several saturated and 1,2-dehydropyrrolizidine alkaloids were detected, mainly as their N-oxides, in fresh pollen collected from flowers of the pyrrolizidine alkaloid-producing plants Echium vulgare, E. plantagineum, Senecio jacobaea, S. ovatus, and Eupatorium cannabinum, and/or pollen loads from bees (bee pollen) that foraged on those plants. A major alkaloidal metabolite in S. ovatus was tentatively identified, using its mass spectrometric data and biogenic considerations, as the previously unreported, saturated alkaloid, 2-hydroxysarracine. Heating had very little effect on the 1,2-dehydropyrrolizidine alkaloids and their N-oxides from a variety of sources. Considered in conjunction with international concerns about the adverse effects of these alkaloids, the results strongly indicate a need for monitoring pollen supplies intended for human consumption, at least until conditions for processing and/or selection are clearly defined such as to significantly reduce the hepatotoxic (and potentially carcinogenic and genotoxic) pyrrolizidine alkaloid content of bee pollen.

Effect of magnesium on iron and magnesium metabolism in rats were investigated. 96 male Wistar rats were divided into four groups received 2.5; 5.0 and 10.0 mg magnesium daily per kg of body weight--dissolved in 2%--solution of arabic gum (tests groups) or clear 2%--solution of arabic gum (test group) for 4 weeks and the next 4 weeks without supplements. Iron concentrations increased in the brain and kidney of the experimental rats, but decreased in the spleen, intestine and liver (2 and 4 weeks only) also in the heart and femur (only 8 week). Percentage of iron retention decreased during the whole experiment. Magnesium concentrations increased in the spleen, liver and intestine of rats. It was shown that at 8 weeks of experiment the magnesium level of heart and femur decreased (only groups received 2.5 mg and 5.0 mg Mg/kg b.w./24 h), but in group received 10.0 mg Mg/kg b.w./24 h increased for all experiment. The apparent retention of magnesium increased in start of the experiment. This results show that oral magnesium supplementation disturbs metabolism of these elements, especially balance of iron.

The mechanisms by which thyroid hormones increase hepatic 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase mRNA levels were investigated in hypophysectomized rats. Feeding these rats a diet supplemented with 0.5% desiccated porcine thyroid powder resulted in a 5-fold increase in the rate of transcription of the HMG-CoA reductase gene as measured by in vitro "run-on" transcription assays in isolated rat liver nuclei. Time courses of change in reductase mRNA, showing the kinetics of approach to new steady-state levels, indicate that reductase mRNA is also 4-6-fold more stable in thyroid hormone-treated animals than in non-treated animals. Reductase mRNA decayed with a half-life of 2.5 h when mevinolin, a potent inhibitor of HMG-CoA reductase, and colestipol, a bile acid sequesterant, were removed from the diet of hypophysectomized rats. When these drugs were removed from the diet of thyroid hormone-treated hypophysectomized rats, reductase mRNA decayed with a half-life of 15 h. Treating rats with only mevinolin and colestipol increased reductase mRNA levels without stabilizing the mRNA. Administration of cycloheximide to thyroid hormone treated rats rapidly decreased HMG-CoA reductase mRNA levels by destabilizing reductase mRNA and decreasing reductase gene transcription. Cycloheximide treatment had no effect on beta-actin gene transcription or steady state levels of beta-actin mRNA. These results suggest that a short-lived protein(s) may mediate the transcriptional and post-transcriptional effects of thyroid hormones on HMG-CoA reductase mRNA levels.

10. Toxicol Lett. 1980 Aug;6(3):125-30. doi: 10.1016/0378-4274(80)90179-4. The mutagenicity of butadiene towards Salmonella typhimurium. de Meester C, Poncelet F, Roberfroid M, Mercier M. Gaseous butadiene (BUT) was mutagenic towards S. typhimurium strain TA 1530 when the incubation mixture was supplemented with a NADPH-fortified rat liver microsomal preparation; mutagenicity increased with the dose. A significant mutagenic effect was similarly observed when the petri dishes, containing the bacteria but no metabolic activation system, were incubated in the presence of butadiene, in a desiccator in which plates containing the S-9 rat liver fraction had been placed. This indirect mutagenic effect was attributed to the formation, by the S-9 mix, of volatile intermediate(s) that migrated and induced mutations in neighbouring bacteria.

INTRODUCTION: Feed additives are widely used to enhance feed efficiency and promote animal growth and health. Broccoli extract, a plant-derived additive rich in bioactive compounds, has potential physiological regulatory effects. However, its specific impact on cattle remains unclear. METHODS: This study investigated the effects of broccoli extract supplementation on growth performance, rumen microbial composition, blood metabolites, and gene expression in the liver and adipose tissue of castrated Holstein bulls. Animals were randomly assigned to three groups and supplemented daily with 0 g, 15 g, or 18 g of broccoli extract for 45 days. RESULTS: No significant differences were observed among groups in average daily gain, dressing percentage, or fecal score (P > 0.05). However, broccoli extract supplementation significantly improved feed intake, lying time, rumination rate, and net meat yield, while reducing subcutaneous fat percentage (P < 0.05). 16S rRNA sequencing revealed increased rumen microbial diversity in the 18 g group. Blood metabolomics showed elevated prostaglandin E2 levels and enrichment in pathways related to inflammation and lipid metabolism. Transcriptomic analysis revealed enrichment of pathways associated with immune responses and lipid regulation. Integrated multi-omics analysis further demonstrated strong correlations between lipid-related metabolites and gene expression patterns. CONCLUSION: Broccoli extract supplementation modulated feeding behavior and rumen microbiota, improved carcass traits, and influenced lipid metabolism and inflammation-related pathways in Holstein cattle. These findings highlight its potential as a functional feed additive for improving beef cattle production.

BACKGROUND: Cellular senescence (CS) is a hallmark of ageing and age-related diseases. While dietary interventions are often explored to reduce CS, less is known about dietary ingredients that induce it. This study systematically reviews the evidence on dietary ingredients that promote CS in animal models and humans. METHODS: Following PRISMA guidelines (PROSPERO: CRD42022338885), PubMed and Embase were searched for studies on dietary ingredients administered via the gastrointestinal tract affecting CS markers in animal models or adults. Risk of bias was assessed using SYRCLE's and Cochrane's tools. RESULTS: From 10,806 articles, 80 studies (77 animal, 3 human) were included. In animals, high-fat diets commonly induced CS across tissues. The plant extract Teng Long Bu Zhong Tang and certain bioactives promoted CS in tumor tissues, potentially offering anti-cancer benefits. Excessive ethanol intake caused CS in the liver and other organs. In humans, increased CS load was linked to red meat-based meals, high protein intake, and DHA-enriched fish oil. Most studies showed unclear risk of bias. CONCLUSIONS: High-fat diets and anti-cancer natural products promote CS in animal models. Preliminary human evidence suggests similar effects from high-protein, red meat-based diets, or DHA-enriched fish oil. Further research is needed to clarify mechanisms and guide dietary and public health recommendations.

The addition of β-agonists to animal feed can significantly improve the lean-meat rate of pigs, cattle, sheep, and other animals. However, the food residues of β-agonists are harmful to human health. When meat with β-agonist residues is consumed, poisoning symptoms such as palpitation, dizziness, and muscle tremors may develop, and damage to the cardiovascular system, liver, and kidney may occur. In this study, a method based on ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was established for the rapid detection of 14 β-agonists (clenbuterol, salbutamol, ractopamine, clorprenaline, terbutaline, tulobuterol, bromobuterol, bambuterol, zilpaterol, mabuterol, fenoterol, arformoterol, cimaterol, and cimbuterol) in animal food sources. The sample pretreatment method and chromatographic conditions were optimized. The samples were hydrolyzed with β-glucuronidase hydrochloride/aryl sulfate esterase in ammonium acetate buffer (pH 5.2). Enzymatic hydrolysis was performed in a constant-temperature water bath ((36±2) ℃) oscillator for 16 h. The samples were cooled to room temperature and extracted with 0.5% formic acid acetonitrile. NaCl was added to separate the organic and aqueous phases, and 5 mL of the upper organic layer was purified using a one-step purification solid-phase extraction column. After drying with nitrogen at 50 ℃, the residue was dissolved in 0.4 mL of 0.2% formic acid aqueous solution. The samples were passed through a 0.22 μm filter and detected by UHPLC-MS/MS with gradient elution using acetonitrile and 0.1% formic acid aqueous solution as the mobile phases. The analytes were separated on a Phenomenex Kinetex F5 column and detected by positive-ion scanning in multiple-reaction monitoring (MRM) mode. Internal and external standard methods were used for quantitative analysis. The effects of the extract pH, solid-phase extraction column, purification method, and dissolved solution on the extraction efficiency were optimized during pretreatment. UHPLC-quadrupole time-of-flight MS was used to verify the purification effect of the one-step purification solid-phase extraction column, and the results indicated that this t

The frequency of some major mycotoxins in marker tissues (liver and kidney) and in muscle tissue of slaughter pigs and cattle, obtained from registered abattoirs in South Africa, was studied. Samples of each three bovine carcasses were obtained from two abattoirs, and samples of three porcine carcasses were from a third abattoir. All samples originated from animals from subsistence farming. All samples were analysed for aflatoxins (AFB1, AFB2, AFG1, AFG2, deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN) using immunoaffinity chromatography extract cleanup and high-performance liquid chromatography (HPLC). At a limit of quantification (LOQ) of 1 μg/kg (individual AFs, 100 μg/kg (DON), 1 μg/kg (OTA) and 20 μg/kg (ZEN)), no mycotoxins were detected in any of the samples.

Hepatitis E virus (HEV) is a biological hazard that must be controlled and is a recognized etiological agent in viral hepatitis. This is a zoonotic virus and can be transmitted through the fecal-oral route. The pig is an important reservoir host of HEV, and is a source of contamination for the consumer after the consumption of raw or undercooked pork products. When detected, the most prevalent genotype of HEV in China is genotype 4 (denoted as HEV-4). To ensure the safety of this food of animal origin, we undertook a survey of HEV contamination in pig livers and pork samples available for sale, in retail outlets in selected cities in China. Viral RNA was purified from samples collected by lysing in Trizol followed by purification using trichloromethane and virus RNA extract kit. An additional step was applied to improve the recovery rate by adding RNase OUT when extracting virus RNA from pig livers, and the RNA productions were washed in 75% (v/v) ethanol to remove inhibitors. In total, 158 pig livers and 80 pork samples were procured and analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). After purification of total RNA from all samples taken and analyzed by RT-qPCR, a single pig liver was positive by this method for HEV. The positive rate was calculated as 0.63%. In this study, a single positive sample was detected. Considering the dietary habits of Chinese people, pork is a popular food that on occasion may be contaminated with HEV, thereby posing a threat to consumer health. Ongoing surveillance is required to assess the risk to human health arising from HEV-contaminated pork being offered for sale, at retail outlets, especially in the areas of China where pig production is practiced.

Alkali-heat DNA extraction, a rapid and economical method, was evaluated for use in the detection of Shiga toxin-producing Escherichia coli in food using real-time PCR assays. Alkali-heat DNA extracts led to highly sensitive detection (102-104 CFU/mL) of stx and O-antigen genes in beef liver, ground beef, sliced pork, cheese, lettuce, radish sprouts, tomato, and spinach, equivalent to the sensitivity obtained using a commercial DNA extraction kit that utilizes proteinase K lysis, and silica membrane purification. Although there were differences in DNA concentration and purity between DNA extraction methods, the sensitivity of real-time PCR assays was similar. These results indicate that alkali-heat DNA extraction is a viable method when testing food products with real-time PCR assays for the presence of stx and O-antigen genes.

The effect of the addition of seaweed extracts from three brown algae species [Ascophyllum nodosum (AN), Fucus vesiculosus (FV) and Bifurcaria bifurcata (BB)], which are a great source of natural antioxidants, on the oxidative stability of refrigerated low-fat pork liver pâtés was studied. In the studied pâtés, half of pork fat was replaced with a mixture consisting of canola and high-oleic sunflower oil (75:25, v/v), thus improving their fatty acid profile in terms of polyunsaturated fatty acids (PUFA). In order to avoid the oxidation of PUFA in the new samples, seaweed extracts (500 ppm) were added. In addition, some samples were formulated with a synthetic antioxidant (BHT at 50 ppm) (BHT) and a control batch (CON) (without antioxidant) was also prepared, for comparison purposes. Thus, in total, five batches of liver pâté were prepared: CON, BHT, AN, FV and BB. Pâté samples were analyzed at 0, 45, 90, 135 and 180 days of refrigerated storage at 4 °C. The addition of seaweed extracts did not modify significantly (P > 0.05) the chemical composition or microbial characteristics of healthy pork liver pâté, except for the protein content, which resulted in a significant increase (≈2-3%) in the batches manufactured with addition of seaweed extracts compared to control samples. At the end of storage (180 days), L* values were significantly lower in the FV and BB batches than in the other batches. Moreover, the a* and b* values were also significantly lower in CON batches than in the samples with antioxidants added. Differences in oxidative parameters (conjugated dienes, TBARs and carbonyls) among batches were observed at the end of the storage time, showing samples with seaweed extracts a similar degree of protection against oxidation compared to BHT formulated samples. A decline of the volatile compounds was noted in all the batches during storage. The total volatile compounds at the end of the storage were significantly lower in BTH, AN, or BB batches than in control batches.

Hepatitis A virus (HAV) can cause serious liver disease and even death. HAV outbreaks are associated with the consumption of raw or minimally processed produce, making it a major public health concern. Infections have occurred despite the fact that effective HAV vaccine has been available. Development of a rapid and sensitive HAV detection method is necessary for an investigation of an HAV outbreak. Detection of HAV is complicated by the lack of a reliable culture method. In addition, due to the low infectious dose of HAV, these methods must be very sensitive. Current methods rely on efficient sample preparation and concentration steps followed by sensitive molecular detection techniques. Using green onions which was involved in most recent HAV outbreaks as a representative produce, a method of capturing virus particles was developed using carboxyl-derivatized magnetic beads in this study. Carboxyl beads, like antibody-coated beads or cationic beads, detect HAV at a level as low as 100 pfu/25g of green onions. RNA from virus concentrated in this manner can be released by heat-shock (98°C 5min) for molecular detection without sacrificing sensitivity. Bypassing the RNA extraction procedure saves time and removes multiple manipulation steps, which makes large scale HAV screening possible. In addition, the inclusion of beef extract and pectinase rather than NP40 in the elution buffer improved the HAV liberation from the food matrix over current methods by nearly 10 fold. The method proposed in this study provides a promising tool to improve food risk assessment and protect public health. Published by Elsevier B.V.

19. Indian J Exp Biol. 2017 Mar;55(3):142-50. Dietary ginger improves glucose dysregulation in a long-term high-fat high-fructose fed prediabetic rat model. Saravanan N, Patil MA, Kumar PU, Suryanarayana P, Reddy GB. The rapid increase in global diabetes burden with its associated morbidity and mortality is a major health concern for humans. Prediabetes is a condition which predispose a person not only to diabetes but also to the associated complications including morbidity even in the absence of an apparant hyperglycemia. However, appropriate dietary intervention may not only prevent but also improve one’s condition as diet is the major contributor to such metabolic disorders. Here, we investigated the effect of dietary ginger (Zingiber officinale Roscoe) on the markers of insulin resistance and pathophysiology in a diet-induced prediabetic rat model. Male Sprague-Dawley (SD) rats were fed the following diets: control (5% groundnut oil + 65 % corn starch), high fat high fructose (HFHF; 25% beef tallow + 35 % fructose) and HFHF with 3 % ginger (HFHFG) for eight months. Plasma markers of insulin resistance, lipid profile, oral glucose tolerance (OGTT; 2nd and 5th month), intraperitoneal insulin tolerance (ITT), plasma total antioxidant capacity (TAC), liver histology and pancreatic immunohistochemistry (IHC) were examined. The impaired OGTT, ITT and insulin sensitivity indices with observed hyperinsulinemia and hypertriglyceridemia suggest that HFHF feeding resulted in prediabetes in rats. HFHF feeding also decreased insulin secretion in the pancreas, increased lipid accumulation in liver and total oxidants in plasma. The effects of HFHF feeding on glucose regulation, pathophysiology of pancreas and liver; total oxidative stress were improved by ginger feeding. The present study demonstrated thatlong-term HFHF feeding induces prediabetes in experimental rats while dietary ginger neutralizes the HFHF induced impairment in glucose regulation, dyslipidemia, and oxidative stress.

A simple method is described for the determination of chloramphenicol and its succinate esters in food products. Examination of food products using high-performance liquid chromatography/high-resolution mass spectrometry showed the presence not only of chloramphenicol but also of its succinate forms. A scheme is proposed for determining chloramphenicol and its succinate esters (calculated as chloramphenicol) in meat (beef, pork, poultry), milk, liver, kidney, eggs, fish and honey. Analytes are extracted from a 1.0 g sample with 5 ml acetonitrile. It was found that using the method of standard addition and diluting the extract with water leads to the elimination of matrix effects and also eliminates errors associated with peak splitting due to the separate elution of the differing forms of the analyte. Validation results were satisfactory, with recoveries from 85% to 111% (meat, milk, liver, kidney, eggs, fish and honey) and a relative standard deviation (RSD) lower than 13% for spiked levels of 0.3, 1.0 and 5 µg kg-1. The limits of detection and quantification (calculated as chloramphenicol for all forms) were 0.1 and 0.3 µg kg-1, respectively. The RSD of the results of the analysis was < 10%. The duration of the analysis was less than 1 h.

Extensive research, intervention equipment, money, and media coverage have been directed at controlling Escherichia coli O157:H7 in beef cattle. However, much of the focus has been on controlling this pathogen postcolonization. This study was conducted to examine the performance, health, and shedding characteristics of beef calves that were vaccinated with an E. coli O157:H7 SRP bacterial extract. These calves had been born to cows vaccinated prepartum with the same vaccine. Cows and calves were assigned randomly to one of four treatments: (i) neither cows nor calves vaccinated with E. coli O157:H7 SRP (CON), (ii) cows vaccinated with E. coli O157:H7 SRP prepartum but calves not vaccinated (COWVAC), (iii) calves vaccinated with E. coli O157:H7 SRP but born to cows not vaccinated (CALFVAC), (iv) cows vaccinated with E. coli O157:H7 SRP prepartum and calves also vaccinated (BOTH). Calves born to vaccinated cows had significantly higher titers of anti-E. coli O157:H7 SRP antibodies (SRPAb) in circulation at branding time (P < 0.001). Upon entry to the feedlot, overall fecal E. coli O157:H7 prevalence was 23 % among calves, with 25 % in the CON treatment group, 19 % in the CALFVAC group, 32 % in the COWVAC group, and 15 % in the BOTH group (P > 0.05). Fecal shedding of E. coli O157 on arrival to the feedlot was not correlated with fecal shedding at slaughter (Spearman's rho = -0.02; P = 0.91). No significant effects of cow or calf E. coli O157:H7 SRP vaccination treatment were found on feedlot calf health or performance (P > 0.05), prevalence of lung lesions or liver abscess (P > 0.05), or morbidity, retreatment, or mortality numbers (P > 0.05). The findings of this study indicate that the timing of vaccination of calves against E. coli O157:H7 may be an important consideration for maximizing the field efficacy of this vaccine.

The Bow Creek watershed (Nebraska, USA) is dominated by the production of beef cattle and row crops; therefore, surface waters are likely to receive runoff containing steroid hormones and pesticides. The goal of the present study was to determine the occurrence and endocrine effects of agrichemicals in this watershed. To accomplish this, four sites within the watershed-Pearl, Bow, and East Bow Creeks and a site at the confluence with the Missouri River-were selected. In June of 2008, polar organic chemical integrative samplers (POCIS) were deployed at each site, whereas in June of 2009, water and sediment samples were collected. Caged fathead minnows (Pimephales promelas) were deployed at all of the selected sites in both years. Analysis of these samples revealed that steroid hormones were not present; however, pesticides were present in POCIS extracts and water samples. In general, the amount of pesticides was higher in POCIS retrieved from Pearl and Bow Creeks than in POCIS from East Bow Creek and the confluence. This variation between sites appeared to be related to row crop density, as row crop land cover surrounding the Pearl and Bow Creek sites was higher than that surrounding the East Bow and confluence sites. To determine the endocrine effects of agrichemicals within this watershed, the hepatic mRNA expression of vitellogenin and estrogen receptor α (ERα), as well as the gonadal expression of P450 aromatase A, was determined for the caged minnows. Females deployed at East Bow Creek and the confluence experienced decreases in the expression of ERα, suggesting that these females had been defeminized; however, this defeminization could not be attributed to any of the pesticides detected at these sites.

The goal of the current study was to determine whether sediments from agriculturally intense watersheds can act as a potential source of anti-estrogenic endocrine-disrupting compounds. The specific objectives of the current study were to determine (1) whether female fathead minnows (Pimephales promelas) experience alterations in endocrine function when exposed to sediments collected from agriculturally intense watersheds and (2) if these sediments display anti-estrogenic activity in an in vitro assay. In addition, sediment samples were analyzed for the presence of steroid hormones and pesticides associated with local agricultural practices. To accomplish this, sediments and water were collected from three sites within two agriculturally intense Nebraska watersheds (Bow Creek and the Elkhorn River). In 2009, minnows were exposed to sediment and/or water collected from the two Bow Creek sites (East Bow Creek and the Confluence) in the laboratory, while in 2010, minnows were exposed to sediment and/or water from East Bow Creek, the Confluence and the Elkhorn River. Following the 7-day exposure period, the hepatic mRNA expression of two-estrogen responsive genes, estrogen receptor α (ERα) and vitellogenin (Vtg) was determined. In 2009, females exposed to Confluence sediments, in the presence of laboratory water or Confluence water, experienced significant reductions in ERα expression relative to unexposed and Confluence water-exposed females. The defeminization of these females suggests the presence of a biologically available anti-estrogenic compound in sediments collected from this site. In 2010, sediments were assessed for anti-estrogenic activity on days 0 and 7 of the exposure period using a 4-h yeast estrogen screen. Lipophilic extracts (LEs) of day 0 sediments collected from the Confluence and the Elkhorn River induced significant reductions in the estrogenic reporter activity of treated yeast cultures suggesting the presence of a lipophilic anti-estrogenic compound in these extracts. Chemical analysis revealed the presence of a variety of steroid hormones, including those associated with the production of beef cattle (i.e. β-trenbolone, α-zearalanol and α-ze

An efficient extraction and cleanup technique, and an instrumental detection method suitable for determination of trace amounts of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and its nitroso-metabolites in animal liver tissue were developed and validated in this paper. The method includes the extraction of explosives from liver tissue samples using accelerated solvent extraction (ASE) followed by cleanup using florisil and styrene-divinyl benzene (SDB) cartridges to remove interfering naturally endogenous compounds. The instrumental analysis was conducted using a capillary column gas chromatograph coupled with an electron capture detector (GC-ECD). High recoveries (58.9-106.8%) of RDX, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX) were achieved at all concentrations studied. RDX, MNX, and TNX gave higher recoveries than DNX at all three tested concentrations (50, 250, 1250 ng/g). Overall recoveries of RDX, MNX, DNX, and TNX from 1g beef liver samples containing 50, 250, and 1250 ng/g were 80.1, 82.8, 68.9, and 80.4%, respectively. The optimal injection port temperature range was 160-170 degrees C for analysis of RDX and its nitroso-metabolites. Higher or lower temperatures than 160-170 degrees C decreased signal amplitudes. RDX was unstable in the liver extraction matrix; as much as 50% of RDX was degraded 10 days after extraction if keeping the liver sample extracts at room temperature. Degradation of RDX to MNX, DNX, or TNX was not detected during the sample storage, extraction, or instrument analysis processes. Other optimized extraction and GC conditions are also discussed.

Hyphenated high-performance liquid chromatography (HPLC)-thermal lens spectrometry (TLS) was demonstrated as a method capable of selectively profiling carotenoids in animal livers and suitable for ultrasensitive determination of trans-beta-carotene in the same tissues. The new approach, HPLC-TLS, a first choice method when studying the tissues containing minute quantities of trans-beta-carotene such as in the case of rat liver, also proved useful in the analysis of beef liver. Due to high sensitivity of the HPLC-TLS method, there is no need for enrichment of liver extracts; in fact, multifold diluted extracts can be analysed directly which not only preserves carotenoids from structural transformations, but also minimises the risk for contamination of the stationary phase. Furthermore, both the calibration in a matrix-matched medium and standard addition method have thus become unnecessary. Reliable determinations of trans-beta-carotene in rat and beef livers (recoveries 98.0 and 99.8%) were obtained by calibration in the mobile phase within the linearity range 1-130 ng ml(-1). In addition, favourable detection limits (0.39 and 0.49 ng ml(-1)), precision of determination (2.8-7.2%) and the selectivity evidenced in the presence of multifold molar excess of vitamins A and E, cholesterol and lecithin, confirm the suitability of the method. The HPLC-TLS approach can be successfully applied in studies concerned with the biotransformation of beta-carotene which are of both scientific and public health interest.