Immersion of debrided diabetic foot ulcer tissue in electrochemically generated pH neutral hypochlorous acid significantly reduces the microbial bioburden: whole-genome sequencing of Staphylococcus aureus, the most prevalent species recovered.

BACKGROUND: Diabetic foot ulcer infections (DFUIs) are the leading cause of lower-limb amputations, mediated predominantly by Staphylococcus aureus. pH-neutral electrochemically generated hypochlorous acid (anolyte) is a non-toxic, microbiocidal agent with significant potential for wound disinfection. AIMS: To investigate both the effectiveness of anolyte for microbial bioburden reduction in debrided ulcer tissues and the population of resident S. aureus. METHODS: Fifty-one debrided tissues from 30 people with type II diabetes were aliquoted by wet weight and immersed in 1- or 10-mL volumes of anolyte (200 parts per million) or saline for 3 min. Microbial loads recovered were determined in colony forming units/g (cfu/g) of tissue following aerobic, anaerobic and staphylococcal-selective culture. Bacterial species were identified and 50 S. aureus isolates from 30 tissues underwent whole-genome sequencing (WGS). FINDINGS: The ulcers were predominantly superficial, lacking signs of infection (39/51, 76.5%). Of the 42/51 saline-treated tissues yielding ≥105 cfu/g, a microbial threshold reported to impede wound-healing, only 4/42 (9.5%) were clinically diagnosed DFUIs. Microbial loads from anolyte-treated tissues were significantly lower than saline-treated tissues using 1 mL (1065-fold, 2.0 log) and 10 mL (8216-fold, 2.1 log) immersion volumes (P<0.0005). S. aureus was the predominant species recovered (44/51, 86.3%) and 50 isolates underwent WGS. All were meticillin susceptible and comprised 12 sequence types (STs), predominantly ST1, ST5 and ST15. Whole-genome multi-locus sequence typing identified three clusters of closely related isolates from 10 patients indicating inter-patient transmission. CONCLUSIONS: Short immersions of debrided ulcer tissue in anolyte significantly reduced microbial bioburden: a potential novel DFUI treatment.

NSAIDs and Renal Impairment: Deprescribing Chronic NSAID use in General Practice.

Aim NSAIDs are high-risk medicines that can commonly cause adverse renal effects. Recent evidence suggests a rise in the number of patients with acute and chronic renal disease. The aim of this audit is to determine our de-prescribing rate of chronic NSAID use in an Irish general practice. Methods We reviewed NSAID-containing drug prescriptions that were issued over a three month period in 2018. A description analysis was performed to ascertain for the frequency and type of NSAIDs prescribed. An educational session was delivered to clinicians to encourage de-prescribing of NSAIDs if deemed clinically appropriate. Results Fifty-one NSAID-containing prescriptions were identified. Thirty-six (71%) patients, who were prescribed a regular NSAID, were aged between 71-85 years. Meloxicam was used the most (31%), whilst the preferred NSAIDs (naproxen and ibuprofen) were used least (18%). A 37% improvement in de-prescribing of chronic NSAIDs was achieved upon re-auditing. Conclusion NSAIDs are commonly implicated in inappropriate prescribing. Clinicians are encouraged to practice de-prescribing at every opportunity. Recent evidence suggests that pharmacy-led educational interventions can further assist de-prescribing of inappropriate medicines. Thus, a close collaboration between physicians and pharmacists is encouraged to further maximise quality of prescribing and patient care.

Hypercalcaemia due to Hypervitaminosis D in a Self-Supplementing Multiple Sclerosis Patient: A Case Report

There is an increasing literature with regard to vitamin D supplementation in Multiple Sclerosis (MS). We report the case of a 45 year-old male with MS who presented with symptomatic hypercalcaemia secondary to self-supplementation of vitamin D3 purchased online. Treatment was with IV hydration, glucocorticoids, calcitonin and bisphosphonates. This case highlights a lack of consensus guidance regarding safe vitamin D supplementation dosages and the importance of a thorough history with regard to non-prescribed supplements, particularly those easily available online.

Sexual selection drives the evolution of male wing interference patterns.

The seemingly transparent wings of many insects have recently been found to display unexpected structural coloration. These structural colours (wing interference patterns: WIPs) may be involved in species recognition and mate choice, yet little is known about the evolutionary processes that shape them. Furthermore, to date investigations of WIPs have not fully considered how they are actually perceived by the viewers' colour vision. Here, we use multispectral digital imaging and a model of Drosophila vision to compare WIPs of male and female Drosophila simulans from replicate populations forced to evolve with or without sexual selection for 68 generations. We show that WIPs modelled in Drosophila vision evolve in response to sexual selection and provide evidence that WIPs correlate with male sexual attractiveness. These findings add a new element to the otherwise well-described Drosophila courtship display and confirm that wing colours evolve through sexual selection.

Children's experiences following a CBT intervention to reduce dental anxiety: one year on.

Objective: To investigate children's ongoing experiences of dental care and use of strategies to manage their dental anxiety following cognitive behavioural therapy (CBT). Design: A child self-completed postal questionnaire. Settings: Hospital, community and general dental practice. Subjects: Questionnaires were sent to 44 children, aged 10­17 years who had been referred to specialist services due to their dental anxiety. Intervention: Children had all previously received a guided CBT self-help intervention to reduce their dental anxiety and, on completion of treatment, had been discharged to their referring dentist. Questionnaires were sent out 12­18 months later to ascertain dental attendance patterns and application of any strategies learnt from the previous CBT intervention. Results: 22 responses (50%) were received from 16 girls and six boys. Eighty-two percent had subsequently accessed follow-up care with a general dental practitioner and over half of these had undergone a dental procedure, other than a check-up. Ninety-one percent reported feeling less worried about dental visits, than previously, and described a change in cognition, behaviours, and feelings that allowed them to manage their anxiety better. Conclusions: CBT has positive immediate and longitudinal effects in reducing children's dental anxiety. The challenge of adopting this evidence-based approach within primary care settings remains.

Lack of phototoxicity potential with delafloxacin in healthy male and female subjects: comparison to lomefloxacin.

AIMS: Delafloxacin is a fluoroquinolone antibiotic recently approved by the FDA for treatment of acute bacterial skin and skin structure infections (ABSSSI). Delafloxacin was assessed for phototoxicity potential compared with a known phototoxic fluoroquinolone. METHODS: A Phase 1, investigator-blind, placebo/active-controlled, randomized, parallel-group study was conducted in 52 healthy male and female volunteers who received 200 or 400 mg of oral delafloxacin, 400 mg oral lomefloxacin or placebo once daily for 6 days. This study evaluated the photosensitizing potential and possible wavelength dependency of delafloxacin by comparing the response of the skin to ultraviolet A (UVA), ultraviolet B (UVB) and visible radiation prior to and during administration of delafloxacin, lomefloxacin as a positive control, or placebo. Adverse events were monitored throughout the study. RESULTS: Forty-seven subjects completed six days of dosing, and no evidence of phototoxicity was seen with delafloxacin. Delafloxacin at 200 and 400 mg day-1 and placebo did not demonstrate differences in percent change from baseline in minimal erythema dose at all tested wavelengths (295-430 nm) by monochromator and solar simulator. Lomefloxacin, the positive control, had statistically significant differences (p < 0.05) at UVA wavelengths of 335 and 365 ± 30 nm 24 hours after radiation exposure (maximum response). The phototoxic index results were significantly higher for lomefloxacin at 335 nm and 365 nm compared to placebo and delafloxacin. CONCLUSIONS: 200 and 400 mg of delafloxacin administered for 6 days were well tolerated in healthy adult volunteers. Delafloxacin and placebo failed to demonstrate a phototoxic effect but lomefloxacin, the positive control, demonstrated moderate phototoxicity.

Antibacterials dispensed in the community comprise 85%-95% of total human antibacterial consumption.

WHAT IS KNOWN AND OBJECTIVE: Interventions intended to slow the emergence and spread of antibacterial resistance through enhanced antimicrobial stewardship will be more effective if informed by an accurate knowledge of current patterns of antibacterial consumption. For example, knowledge of the relative magnitude of community antibacterial consumption in relation to hospital antibacterial consumption within each nation or region should help guide decisions about the relative importance of community and hospital antimicrobial stewardship programmes. It is commonly stated that community antibacterial consumption comprises approximately 80% of total national antibacterial consumption. We aimed to determine this proportion across a large range of nations. METHODS: We measured community and hospital antibacterial consumption in New Zealand during 2015, from both reimbursement and purchase data, and compared the New Zealand data with those reported from a large range of other nations during similar time periods. RESULTS AND DISCUSSION: Community antibacterial consumption comprised approximately 85%-95% of total antibacterial consumption in all nations for which data were available, and in New Zealand comprised a higher proportion than in any other nation. The proportion of total antibacterial consumption comprised by community consumption was significantly higher in countries with relatively high levels of total antibacterial consumption than in countries with relatively low levels of total antibacterial consumption. WHAT IS NEW AND CONCLUSION: The high proportion of total antibacterial consumption comprised by community antibacterial consumption suggests devoting particular attention to improved community antimicrobial stewardship. These results suggest that improving antimicrobial stewardship in the community may provide greater overall benefits in combating antibacterial resistance than improving antimicrobial stewardship in hospitals.

Culturally capable and culturally safe: Caseload care for Indigenous women by Indigenous midwifery students.

BACKGROUND: Evidence is emerging of the benefits to students of providing continuity of midwifery care as a learning strategy in midwifery education, however little is known about the value of this strategy for midwifery students. AIM: To explore Indigenous students' perceptions of providing continuity of midwifery care to Indigenous women whilst undertaking a Bachelor of Midwifery. METHODS: Indigenous Bachelor of Midwifery students' experiences of providing continuity of midwifery care to Indigenous childbearing women were explored within an Indigenous research approach using a narrative inquiry framework. Participants were three Indigenous midwifery students who provided continuity of care to Indigenous women. FINDINGS: Three interconnected themes; facilitating connection, being connected, and journeying with the woman. These themes contribute to the overarching finding that the experience of providing continuity of care for Indigenous women creates a sense of personal affirmation, purpose and a validation of cultural identity in Indigenous students. DISCUSSION AND CONCLUSIONS: Midwifery philosophy aligns strongly with the Indigenous health philosophy and this provides a learning platform for Indigenous student midwives. Privileging Indigenous culture within midwifery education programs assists students develop a sense of purpose and affirms them in their emerging professional role and within their community. The findings from this study illustrate the demand for, and pertinence of, continuity of care midwifery experiences with Indigenous women as fundamental to increasing the Indigenous midwifery workforce in Australia. Australian universities should provide this experience for Indigenous student midwives.

Structural Brain Changes following Long-Term 6° Head-Down Tilt Bed Rest as an Analog for Spaceflight.

BACKGROUND AND PURPOSE: Following long-term spaceflight, a subset of the National Aeronautics and Space Administration astronauts present with visual impairment and increased intracranial pressure, known as visual impairment and intracranial pressure syndrome. We investigated structural brain changes following long-term head-down tilt bed rest as a spaceflight analog. MATERIALS AND METHODS: Volumetric analysis was performed on structural pre- and post-bed rest brain MR images. RESULTS: Comparing post-bed rest to pre-bed rest images, we found the following: 1) no significant group differences in GM, WM, CSF, or ventricular volumes; 2) shift of the center of mass of the brain upward and posterior rotation of the brain relative to the skull; 3) a significant correlation between posterior brain rotation and changes in ventricular volume; and 4) significant increases in brain tissue density in regions at the vertex, including the frontoparietal lobes, with contraction of adjacent extra-axial CSF spaces, and significant decreases in tissue density in areas along the base of the brain, including the orbitofrontal cortex. CONCLUSIONS: We observed widespread morphologic changes with brain tissue redistribution in response to gravity changes; possible associated functional changes are unknown. The observation that ventricular change is correlated to posterior brain rotation suggests an alteration in CSF homeostasis. Ultimately, to elucidate any structural changes that may play a role in visual impairment and intracranial pressure syndrome, volumetric analysis of pre- and postflight structural scans of astronauts is needed.

The CLRX.1/NOD24 (NLRP2P) pseudogene codes a functional negative regulator of NF-κB, pyrin-only protein 4.

Pseudogenes are duplicated yet defunct copies of functional parent genes. However, some pseudogenes have gained or retained function. In this study, we consider a functional role for the NLRP2-related, higher primate-specific, processed pseudogene NLRP2P, which is closely related to Pyrin-only protein 2 (POP2/PYDC2), a regulator of nuclear factor-κB (NF-κB) and the inflammasome. The NLRP2P open-reading frame on chromosome X has features consistent with a processed pseudogene (retrotransposon), yet encodes a 45-amino-acid, Pyrin-domain-related protein. The open-reading frame of NLRP2P shares 80% identity with POP2 and is under purifying selection across Old World primates. Although widely expressed, NLRP2P messenger RNA is upregulated by lipopolysaccharide in human monocytic cells. Functionally, NLRP2P impairs NF-κB p65 transactivation by reducing activating phosphorylation of RelA/p65. Reminiscent of POP2, NLRP2P reduces production of the NF-κB-dependent cytokines tumor necrosis factor alpha and interleukin (IL)-6 following toll-like receptor stimulation. In contrast to POP2, NLRP2P fails to inhibit the ASC-dependent NLRP3 inflammasome. In addition, beyond regulating cytokine production, NLRP2P has a potential role in cell cycle regulation and cell death. Collectively, our findings suggest that NLRP2P is a resurrected processed pseudogene that regulates NF-κB RelA/p65 activity and thus represents the newest member of the POP family, POP4.

Intake estimates of naturally occurring phytosterols using deterministic and probabilistic methods in a representative Irish population.

Intakes of naturally occurring phytosterols were estimated in the Irish population using phytosterol concentration data obtained from the published literature. Both deterministic and probabilistic approaches were used. Concentration data were applied to dietary intake data as assessed in the North/South Ireland Food Consumption Survey (n = 1,379). With the probabilistic approach, a uniform distribution was used in place of fixed values to reflect variability and uncertainty in phytosterol concentrations. Foods consumed were also categorized into 19 food groups, and the phytosterol intake per food group was calculated. Using the deterministic approach, mean phytosterol intake for the total population was 254 mg/day (28.1 mg/1,000 kJ). Using the probabilistic approach, slightly higher mean phytosterol intakes of 287 mg/day were found. In terms of food groups, 'bread and bread-rolls', 'vegetables and vegetable dishes' and 'spreading fat and oils' were the top three contributing food groups to phytosterol intakes. This study has shown how both deterministic and probabilistic approaches may be used to determine phytosterol intakes in a population.

Habitual fish consumption does not prevent a decrease in LCPUFA status in pregnant women (the Seychelles Child Development Nutrition Study).

Information on the status of long-chain polyunsaturated fatty acids (LCPUFAs) in pregnancy and breast milk in very high fish-eating populations is limited. The aim of this study was to examine dietary intake and changes in fatty acid status in a population of pregnant women in the Republic of Seychelles. Serum docosahexaenoic acid (DHA) decreased significantly between 28-week gestation and delivery (n=196). DHA status did not correlate significantly with length of gestation and was not associated with self-reported fish intake, which was high at 527 g/week. In breast milk, the ratio of DHA to arachidonic acid (AA) was consistent with those observed in other high fish-eating populations. Overall the data suggest that high exposure to LCPUFAs from habitual fish consumption does not prevent the documented decrease in LCPUFA status in pregnancy that occurs as a result of foetal accretion in the third trimester of pregnancy.

Clinical biochemistry guidelines in Wales: their impact on laboratory services.

BACKGROUND: Evidence of significant variations in clinical biochemistry practice in Wales led to the formation of the All Wales Clinical Biochemistry Audit Group (AWCBAG) in 1993, with the aim of auditing laboratory services to ensure that they are optimally used. As part of this process, clinical guidelines are produced and circulated to all clinical biochemistry departments in Wales. The current aim of the AWCBAG is to assess the extent and impact of adoption of these guidelines across Wales. METHODS: Three surveys were dispatched at intervals over a decade to all clinical biochemistry departments in Wales to investigate practice in: (1) urine albumin testing to screen for diabetic nephropathy; (2) biochemical investigation of menopausal status and the monitoring of hormone replacement therapy; (3) screening for Cushing's syndrome. RESULTS: The results show that laboratories across Wales are generally following guideline criteria and are adapting their practice in-line with changing recommendations. CONCLUSION: The introduction of AWCBAG guidelines has been widely accepted by clinical biochemistry departments in Wales. These guidelines have led to a more efficient and effective use of laboratory services.

A randomised interventional trial of omega-3-polyunsaturated fatty acids on endothelial function and disease activity in systemic lupus erythematosus.

OBJECTIVE: To determine the clinical effect of dietary supplementation with low-dose omega-3-polyunsaturated fatty acids on disease activity and endothelial function in patients with systemic lupus erythematosus. METHODS: A 24-week randomised double-blind placebo-controlled parallel trial of the effect of 3 g of omega-3-polyunsaturated fatty acids on 60 patients with systemic lupus erythematosus was performed. Serial measurements of disease activity using the revised Systemic Lupus Activity Measure (SLAM-R) and British Isles Lupus Assessment Group index of disease activity for systemic lupus erythematosus (BILAG), endothelial function using flow-mediated dilation (FMD) of the brachial artery, oxidative stress using platelet 8-isoprostanes and analysis of platelet membrane fatty acids were taken at baseline, 12 and 24 weeks. RESULTS: In the fish oil group there was a significant improvement at 24 weeks in SLAM-R (from 9.4 (SD 3.0) to 6.3 (2.5), p<0.001); in BILAG (from 13.6 (6.0) to 6.7 (3.8), p<0.001); in FMD (from 3.0% (-0.5 to 8.2) to 8.9% (1.3 to 16.9), p<0.001) and in platelet 8-isoprostanes (from 177 pg/mg protein (23-387) to 90 pg/mg protein (32-182), p = 0.007). CONCLUSIONS: Low-dose dietary supplementation with omega-3 fish oils in systemic lupus erythematosus not only has a therapeutic effect on disease activity but also improves endothelial function and reduces oxidative stress and may therefore confer cardiovascular benefits.

Use of a food-consumption database with packaging information to estimate exposure to food-packaging migrants: expoxidized soybean oil and styrene monomer.

There is no set protocol for completing refined exposure assessments of food-packaging migrants in the European Union. One novel method that could be used to provide more realistic exposure assessments and also reduce uncertainty in the exposure estimation could be the use of food consumption surveys that also have packaging information. The aim of the current study was to estimate exposure to two food-packaging migrants (expoxidized soybean oil (ESBO) and styrene monomer) using a food-consumption database that collected packaging information. The Irish National Children's Food Survey (NCFS) was completed in 2003-04 and it collected information on the type and amount of food consumed by 594 Irish children aged 5-12 years, in addition to the type of packaging used for these foods. The Irish Food Packaging Database (IFPD) was completed in parallel to this food consumption survey and recorded exact information on the contact layer used for the packaging. In a database that combined information from the NCFS and the IFPD, the packaging materials that could contain the target migrants were identified. If a food was packaged in a material that could contain the migrant, it was assumed that the migrant was present in the food. For the exposure assessment of ESBO the 90th percentile migration values of ESBO in foods derived from the literature were used. This was similar to a method as used by the European Food Safety Authority (EFSA) in their exposure assessment of ESBO for adults. Two scenarios of styrene exposure were undertaken in this study. In the first scenario the 90th percentile migration value for styrene found in foods was used; in the second scenario the maximum level of styrene found in foods was used. These migration values were derived from the literature. The mean intake of ESBO for Irish children was 0.023 mg kg-1 body weight day-1, which is well below the tolerable daily intake (TDI) of 1 mg kg-1 body weight day-1 set by the Scientific Committee for Food (SCF) in 1999. The food group that contributed most to ESBO intake was tomato sauces packed in glass jars with polyvinyl chloride (PVC)-lined metal lids (46.8%). For styrene, the mean intake was 0.122 microg kg-1 body weight day-1 when using the 90th percentile migration values and 0.169 microg kg-1 body weight day-1 when using the maximum migration values. These estimated intakes are below the provisional maximum tolerable daily intake (PMTDI) of 40 microg kg-1 body weight day-1, which was established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 1984. Therefore, the estimated intakes of the two migrants are not of concern for Irish Children and uncertainty is reduced in the assessment due to the fact that information is available on the type of foods consumed the type of packaging used for these foods.

Estimation of exposure to food packaging materials. 3: Development of consumption factors and food-type distribution factors from data collected on Irish children.

There are many initiatives in Europe trying to refine the exposure assessment for food packaging migrants. In the USA, the US Food and Drug Administration (US FDA) uses food consumption and food-type distribution factors to aid the exposure assessment process and generate more realistic estimates of exposure. The present study developed food-consumption factors and food-type distribution factors for Irish children aged 5-12 years from data collected as part of a National Children's Food Survey (NCFS) completed in Ireland in 2003-04, combined with data from the 2003-05 Irish Food Packaging Database and from literature data on surface area-to-weight ratios for food packaging. Consumption factors are defined as the fraction of a person's diet likely to contact a specific food-contact material, while food-type distribution factors reflect the fraction of all food contacting each material that is aqueous, acidic, alcoholic or fatty. In this study, it was found that 'total plastics' had the highest consumption factor of 0.83, while 'total paper and board' had a factor of 0.13 and 'total metal and alloys' had a factor of 0.06. Although it is tentative to compare the US FDA factors with the factors derived in the current study, as the US FDA data are for the total US population, the consumption factor for 'total plastics' in the present study (0.83) was similar to that used by the US FDA (0.79). However, a large difference existed for the consumption factor for 'total metal and alloys' in the Irish data (0.06) and the US FDA data (0.2). In terms of the type of materials used for foods, glass was used mainly for acidic foods (0.67), while plastic was uniformly used for all food types. The food-contact area for plastic packaging for all foods consumed by children was 10.67 dm2/child day(-1), which is slightly lower than the proposed value for the average European consumer of 12.4 dm2/person day(-1). However, this should be expected, as children do not consume as much food as adults. When scenario exposure estimates were made, it was found that when using European Union assumptions, estimates were much larger than when using consumption factors and food-type distribution factors in the exposure assessment, even when conservative assumptions were employed for the consumption factor. This study highlights the potential use of consumption factors and food-type distribution factors in the refinement of the exposure assessment process and how these factors can be calculated using data collected as part of a national food survey.

Estimation of exposure to food packaging materials. 1: Development of a food-packaging database.

A food-packaging database was developed to provide qualitative information on the types of packaging materials used for foods. Packaging information was collected from a sample of 594 children aged 5-12 years as part of a national children's food survey carried out in Ireland during 2003-04. All the food packaging collected during the survey was forwarded to the coordinating centre for further analysis and entry into the Irish Food Packaging Database. The database was created in Microsoft Access and stored information on: the brand of the food, the packaging type, the unit weight, the contact layer, the European Union food type (i.e. aqueous, acidic, alcoholic or fatty) and other relevant parameters. Of the 5551 different brand foods consumed by children in the food survey, packaging information was collected on 3441 (62%). As some brand foods had different unit weights and packaging formats, there was duplication of some brand foods in the database to account for this fact. Therefore, there were 3672 packaging entries in the database. Of these, plastics were the most common packaging contact layer (n = 2874, 78.3%). Multimaterial multilayers with a plastic contact layer accounted for 459 (12.5%) entries. Polyethylene was the most frequently used contact layer (n = 941), with polypropylene a close second (n = 809). This database is unique in Europe for the quality and amount of food packaging information it contains and could be used to develop packaging use factors for a more refined exposure assessment to food packaging materials in the European Union.

Estimation of exposure to food-packaging materials. 2: patterns of intakes of packaged foods in Irish children aged 5-12 years.

In the European Union (EU), many assumptions are employed to calculate the intake of migrating chemicals from food packaging. However, very little is known about the actual intake of packaged food, the type of this food and the type of packaging used for this food. The objective of the current study was to examine intakes of packaged food in children aged 5-12 years to provide information on the types of food that are packaged and the type of packaging used. To do this, a food-consumption database, which also recorded information on packaging, was merged with a packaging database, which provided information on the contact layers of packaging. Foods were classified into EU Food categories according to European Council Directive EC 85/575/EEC (European Council 1985), which determined their food type (i.e. aqueous, acidic, alcoholic and fatty). The mean daily intake of all packaged food was 1195 g day-1 with an upper intake of 1959 g day-1 (97.5th percentile); the intake of food packaged in plastic was 993 g day-1 with an upper percentile of 1692 g day-1 (97.5th percentile). The mean daily intake of fat from all packaged food was 62 g day-1, with an upper intake of 100 g day-1 (97.5th percentile). When this was investigated further, it was found that the mean fat intake from packaged 'fatty' foods only was 32 g day-1, with an upper intake of 61 g day-1 (97.5th percentile). The food that contributed most to fat intake was milk. As many food chemical intake assessments are moving towards probabilistic methods, probabilities of a food being packaged and the probability of the type of packaging used were determined. The probability of food being packaged was 0.88. Some foods not 100% packaged included fruit, vegetables, liquid beverages non-alcoholic (includes water) and bread. Probabilities were also derived for the packaging types used for food. It can be concluded that not all the individual assumptions used in the EU exposure assessment are conservative, but in combination they are conservative.

Survival of salmonella transformed to express green fluorescent protein on Italian parsley as affected by processing and storage.

To study the effect of processing and storage parameters on the survival of Salmonella on fresh Italian parsley, parsley bunches were dipped for 3 or 15 min in suspensions that were preequilibrated to 5, 25, or 35 degrees C and inoculated with Salmonella transformed to express enhanced green fluorescent protein. Loosely attached and/or associated, strongly attached and/or associated, and internalized and/or entrapped Salmonella cells were enumerated over 0, 1, and 7 days of storage at 25 degrees C and over 0, 1, 7, 14, and 30 days of storage at 4 degrees C using surface-plating procedures. Leaf sections obtained from samples after 0, 1, and 7 days of storage were examined using confocal scanning laser microscopy. Temperature of the dip suspension had little effect on the attachment and survival of Salmonella cells on parsley. Regardless of the temperature or duration of dip, Salmonella was internalized. Immersion for longer times resulted in higher numbers of attached and internalized cells. Microscopic observations supported these results and revealed Salmonella cells near the stomata and within cracks in the cuticle. Storage temperature had the greatest impact on the survival of Salmonella cells on parsley. When stored at 25 degrees C, parsley had a shelf life of 7 days, and Salmonella populations significantly increased over the 7 days of storage. For parsley stored at 4 degrees C, numbers of Salmonella cells decreased over days 0, 1, and 7. After 7 days of storage, there were no viable internalized Salmonella cells detected. Storage temperature represents an important control point for the safety of fresh parsley.