Non-targeted screening for contaminants in paper and board food-contact materials using effect-directed analysis and accurate mass spectrometry

Due to large knowledge gaps in chemical composition and toxicological data for substances involved, paper and board food-contact materials (P&B FCM) have been emerging as a FCM type of particular concern for consumer safety. This study describes the development of a step-by-step strategy, including extraction, high-performance liquid chromatography (HPLC) fractionation, tentative identification of relevant substances and in vitro testing of selected tentatively identified substances. As a case study, we used two fractions from a recycled pizza box sample which exhibited aryl hydrocarbon receptor (AhR) activity. These fractions were analysed by gas chromatography (GC) and ultra-HPLC (UHPLC) coupled to quadrupole time-of-flight mass spectrometers (QTOF MS) in order tentatively to identify substances. The elemental composition was determined for peaks above a threshold, and compared with entries in a commercial mass spectral library for GC-MS (GC-EI-QTOF MS) analysis and an in-house built library of accurate masses for substances known to be used in P&B packaging for UHPLC-QTOF analysis. Of 75 tentatively identified substances, 15 were initially selected for further testing in vitro; however, only seven were commercially available and subsequently tested in vitro and quantified. Of these seven, the identities of three pigments found in printing inks were confirmed by UHPLC tandem mass spectrometry (QqQ MS/MS). Two pigments had entries in the database, meaning that a material relevant accurate mass database can provide a fast tentative identification. Pure standards of the seven tentatively identified substances were tested in vitro but could not explain a significant proportion of the AhR-response in the extract. Targeted analyses of dioxins and PCBs, both well-known AhR agonists, was performed. However, the dioxins could explain approximately 3% of the activity observed in the pizza box extract indicating that some very AhR active substance(s) still remain to be identified in recycled low quality P&B.     

Effects of infantile undernutrition on adult learning in rats: Methodological and design problems.

Examines research bearing upon possible confounding variables involved in the study of the effects of early undernutrition upon later learning in rats. The procedures currently used to produce undernutrition also produce changes in the infant rats' social and environmental conditions. Evidence from other sources indicate that these types of changes may play an important role in the determination of later behavior. Attribution of the causes of later effects of undernutrition must be reinterpreted in light of this evidence. Early undernutrition and the variables confounded with it produce effects on later size, motor ability, behavior toward food, ability to absorb and use food and to adjust to food deprivation, and responsibility to aversive and environmental stimuli. Thus, the effects of undernutrition upon later learning may be due to the effects upon these other variables which can affect learning performance rather than upon learning capacity. Strategies for dealing with these problems depend upon the goals of the researcher. (86 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Quantification of antibiotic use on dairy farms in Pennsylvania

Antibiotic use data are critical for drawing conclusions about the epidemiological connections between antibiotic use in farms animals, antibiotic resistance, animal health, and human health. The goal of this study was to quantitatively and qualitatively characterize antibiotic use on dairy farms in Pennsylvania, the state with second largest number of dairy farms nationally. A survey was sent to 10% of the 6,580 dairy farms registered in Pennsylvania and completed by 235 producers (response rate of 36%). Data on antibiotic use in the previous month and in the previous 6 mo were collected based on farmer self-report, using either recall or treatment records. Two metrics were used to quantify antibiotic consumption: animal-defined daily doses (ADD) and days of therapy (DOT), a metric used in human medicine for purposes of antimicrobial stewardship. Across all farms, 24,444 ADD and 19,029 DOT were reported, representing treatment incidences of 4.2 ADD/1,000 animal-days and 3.3 DOT/1,000 animal-days. These rates were generally lower than those found in other states and countries. The main indication for antibiotic use was mastitis, and first-generation cephalosporins were the most commonly used class of antibiotic for all indications, followed by penicillins and third-generation cephalosporins. Trends in use were similar for ADD and DOT, but the numbers of recorded DOT and associated treatment incidences were generally lower than the number of ADD and associated treatment incidences. Rates of treatment were significantly associated with herd size. This study is the first to quantify antibiotic use on dairy farms in Pennsylvania and the first to use the DOT metric in a dairy setting.     

Physiological adaptation of Escherichia coli after transfer onto refrigerated ground meat and other solid matrices: A molecular approach

Bacteria on meat are subjected to specific living conditions that differ drastically from typical laboratory procedures in synthetic media. This study was undertaken to determine the behavior of bacteria when transferred from a rich-liquid medium to solid matrices, as is the case during microbial process validation. Escherichia coli cultured in Brain-Heart Infusion (BHI) broth to different growth phases were inoculated in ground beef (GB) and stored at 5°C for 12 days or spread onto BHI agar and cooked meat medium (CMM), and incubated at 37°C for several hours. We monitored cell densities and the expression of σ factors and genes under their control over time. The initial growth phase of the inoculum influenced growth resumption after transfer onto BHI agar and CMM. Whatever the solid matrix, bacteria adapted to their new environment and did not perceive stress immediately after inoculation. During this period, the σ(E) and σ(H) regulons were not activated and rpoD mRNA levels adjusted quickly. The rpoS and gadA mRNA levels did not increase after inoculation on solid surfaces and displayed normal growth-dependent modifications. After transfer onto GB, dnaK and groEL gene expression was affected more by the low temperature than by the composition of a meat environment.     

Survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on Inoculated Almonds and Pistachios Stored at -19, 4, and 24° C

The survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes was determined on almonds and pistachios held at typical storage temperatures. Almond kernels and inshell pistachios were inoculated with four- to six-strain cocktails of nalidixic acid-resistant Salmonella, E. coli O157:H7, or L. monocytogenes at 6 log CFU/g and then dried for 72 h. After drying, inoculated nuts were stored at -19, 4, or 24°C for up to 12 months. During the initial drying period after inoculation, levels of all pathogens declined by 1 to -log CFU/g on both almonds and pistachios. During storage, moisture content (4.8%) and water activity (0.4) of the almonds and pistachios were consistent at -19°C; increased slowly to 6% and 0.6, respectively, at 4°C; and fluctuated from 4 to 5% and 0.3 to 0.5 at 24°C, respectively. Every 1 or 2 months, levels of each pathogen were enumerated by plating; samples were enriched when levels fell below the limit of detection. No reduction in population level was observed at -19 or 4°C for either pathogen, with the exception of E. coli O157:H7-inoculated almonds stored at 4°C (decline of 0.09 log CFU/g/month). At 24°C, initial rates of decline were 0.20, 0.60, and 0.71 log CFU/g/month on almonds and 0.15, 0.35, and 0.86 log CFU/g/month on pistachios for Salmonella, E. coli O157:H7, and L. monocytogenes, respectively, but distinct tailing of the survival curves was noted for both E. coli O157:H7 and L. monocytogenes.     

Effects of breed and age at slaughter on degradation of muscle lipids during processing of dry‐cured hams

Hams from Landrace, Duroc and Hampshire pigs slaughtered at ages 6, 7.5 and 9 months were processed to generate Norwegian Parma‐style hams. Lipid contents and the compositions of fatty acid classes (ΣSFA, ΣMUFA, ΣPUFA) within neutral lipids, phospholipids and free fatty acids were determined. Small differences in lipid degradation and composition of the classes were revealed. However, significant sensory differences related to lipids were observed. Breed was more important than age. Dry‐cured Hampshire hams gave a more intense mature odour that may be associated with higher overall lipid degradation. Unexpectedly, these hams also demonstrated high juiciness and tenderness, which could be related to the melting characteristics of the fat. Dry‐cured Duroc hams showed a higher susceptibility towards rancidity, presumably associated with preferential oxidation of n‐6 fatty acids relative to C18:1 n‐9. Dry‐cured Landrace hams showed the lowest juiciness and tenderness, likely due to their lower fat content (marbling).     

Thermal properties of duck fatty liver (foie gras) products

Thermal properties of duck fatty liver (foie gras), foie gras emulsion and fatty liver fat, as well as regular duck fat, were determined as a function of temperature. Density of foie gras and foie gras fat and emulsion at 20°C was measured as 947, 836, and 928 kg/m³, respectively. Differential scanning calorimetry thermograms for foie gras fat resulted in melting points ranging from –20 to 40°C. Values for the specific heat at 65°C for the fatty liver, its emulsion and fat, and duck fat were 1.79, 2.38, 1.71, and 2.48 J/g°C, respectively. Thermal conductivity of foie gras (organ) and its emulsion at 40°C was determined as 0.330 and 0.428 W/m°C, respectively. Mathematical models based on composition and temperature were developed for all the thermal properties obtained in this work.     

Effective use of nisin to control Bacillus and Clostridium spoilage of a pasteurized mashed potato product

Heat-resistant spore-forming bacteria such as Bacillus and Clostridium can survive and grow in cooked potato products. This situation represents both a public health problem and an economic problem. The natural food preservative nisin is used in heat-treated foods to prevent the growth of such bacteria. A cocktail of Clostridium sporogenes and Clostridium tyrobutyricum spores was inoculated into cooked mashed potatoes, which were vacuum packed, pasteurized, and incubated at 8 and 25 degrees C. The shelf life of the mashed potatoes at 25 degrees C was extended by at least 58 days with the addition 6.25 microg of nisin per g. At 8 degrees C, in control samples not containing nisin, the natural contaminant Bacillus grew, but the inoculated Clostridium strains did not until the temperature was raised to 20 degrees C after 39 days. No bacterial growth occurred in nisin-containing samples. The shelf life of the mashed potatoes was extended by at least 30 days with 6.25 microg of nisin per g. In trials involving a cocktail of Bacillus cereus and Bacillus subtilis strains, 6.25 microg of nisin per g extended the shelf life of mashed potato samples that were not vacuum packed by at least 27 days at 8 degrees C. At 25 degrees C, 25 microg of nisin per g extended shelf life by a similar period. Shelf life extension was also observed at lower nisin levels. Microbiological analysis of the mashed potato ingredients showed that a high spore level was associated with the onion powder. It is emphasized that the preservative and the ingredients must be well mixed to ensure good nisin efficacy. Nisin remained at effective levels after pasteurization, and good retention was observed throughout the shelf life of the mashed potatoes.