Inactivation of Cryptosporidium parvum oocysts and faecal indicator bacteria in cattle slurry by addition of ammonia.

AIMS: To determine inactivation of Cryptosporidium parvum oocysts and reduction of Escherichia coli and enterococci in cattle slurry added aqueous ammonia. METHODS AND RESULTS: Escherichia coli, enterococci and nonviable C. parvum oocysts (DAPI+PI+) were enumerated every second day for 2 weeks in cattle slurry amended with 60 mmol l-1 aq. ammonia and compared with untreated slurry at three temperatures. Regardless of temperature, the proportion of nonviable C. parvum oocysts increased significantly faster over time in slurry with added ammonia than raw slurry (P = 0·021) corresponding to 62·0% higher inactivation (P = 0·001) at day 14. Additionally, 91·8% fewer E. coli and 27·3% fewer enterococci were observed in slurry added ammonia at day 14 compared to raw slurry. CONCLUSION: The addition of aqueous ammonia to raw slurry significantly reduced the viability of C. parvum oocysts and numbers of bacterial indicators. Hence, ammonia is usable at lower pathogen concentrations in slurry before application to agricultural land. SIGNIFICANCE AND IMPACT OF THE STUDY: Livestock waste is a valuable source of plant nutrients and organic matter, but may contain high concentrations of pathogens like E. coli and Cryptosporidium sp. that can be spread in the environment, and cause disease outbreaks. However, die-off rates of pathogens in organic waste can increase following increasing ammonia concentrations.

Phase II multicentre trial of oral quisinostat, a histone deacetylase inhibitor, in patients with previously treated stage IB-IVA mycosis fungoides/Sézary syndrome.

BACKGROUND: Quisinostat is a hydroxamate, second-generation, orally available pan-histone deacetylase inhibitor. OBJECTIVES: To evaluate the efficacy and safety of oral quisinostat in patients with previously treated cutaneous T-cell lymphoma (CTCL). METHODS: Patients received quisinostat 8 mg or 12 mg on days 1, 3 and 5 of each week in 21-day treatment cycles. Primary efficacy end point was cutaneous response rate (RR) based on the modified Severity Weighted Assessment Tool (mSWAT). Secondary end points included global RR, duration of response (DOR) in skin, progression-free survival (PFS), pruritus relief, safety and pharmacodynamic markers. RESULTS: Eight of 26 (25 evaluable) patients achieved ≥ 50% reduction in mSWAT score at least once, with confirmed cutaneous response in six (RR 24%). There was a low global RR of 8%. DOR in skin ranged from 2·8 to 6·9 months. Median PFS was 5·1 months. Pruritus relief was more frequent in cutaneous responders (67%) than nonresponders (32%). Serial tumour biopsies revealed an increase in acetylated tubulin, indicating a target effect of histone deacetylase 6. Twenty-one of 26 (81%) patients were withdrawn from the study before or at clinical cut-off; five (19%) continued to receive treatment with quisinostat. The most common drug-related adverse events were nausea, diarrhoea, asthenia, hypertension, thrombocytopenia and vomiting. Grade 3 drug-related adverse events included hypertension, lethargy, pruritus, chills, hyperkalaemia and pyrexia. CONCLUSIONS: Quisinostat 12 mg three times weekly is active in the treatment of patients with relapsed or refractory CTCL, with an acceptable safety profile. Combination therapy with other drugs active in CTCL may be appropriate.

Children with cerebral palsy do not achieve healthy physical activity levels.

AIM: This study compared daily activity energy expenditure (AEE) in children with cerebral palsy with a control group and investigated whether the children achieved healthy levels of physical activity. METHODS: We enrolled eight children with bilateral cerebral palsy, from eight to 10 years of age, and a group of controls matched for age and gender. For three days, physical activity was simultaneously measured by accelerometers and self-reports using a diary. The daily AEE results were compared between groups and methods. The number of children that achieved healthy physical activity levels in each group was explored. RESULTS: Children with cerebral palsy had significantly lower daily AEE, as measured by accelerometers, than the controls, and they did not achieve the healthy moderate to heavy physical activity level defined in the Nordic Nutrition Recommendations. Self-reports using the diaries resulted in an overestimation of physical activity compared with the ankle accelerometer measurements in both groups. CONCLUSION: Our investigation of physical activity in children with cerebral palsy and controls using accelerometers and a diary found low levels of daily AEE and physical activity, and these results were most prominent in the group with cerebral palsy. The diaries overestimated physical activity in both groups.

Animal board invited review: Specialising and intensifying cattle production for better efficiency and less global warming: contrasting results for milk and meat co-production at different scales.

Cattle are the world's largest consumers of plant biomass. Digestion of this biomass by ruminants generates high methane emissions that affect global warming. In the last decades, the specialisation of cattle breeds and livestock systems towards either milk or meat has increased the milk production of dairy cows and the carcass weight of slaughtered cattle. At the animal level and farm level, improved animal performance decreases feed use and greenhouse gas emissions per kg of milk or carcass weight, mainly through a dilution of maintenance requirements per unit of product. However, increasing milk production per dairy cow reduces meat production from the dairy sector, as there are fewer dairy cows. More beef cows are then required if one wants to maintain the same meat production level at country scale. Meat produced from the dairy herd has a better feed efficiency (less feed required per kg of carcass weight) and emits less methane than the meat produced by the cow-calf systems, because the intake of lactating cows is largely for milk production and marginally for meat, whereas the intake of beef cows is entirely for meat. Consequently, the benefits of breed specialisation assessed at the animal level and farm level may not hold when milk and meat productions are considered together. Any change in the milk-to-meat production ratio at the country level affects the numbers of beef cows required to produce meat. At the world scale, a broad diversity in feed efficiencies of cattle products is observed. Where both productions of milk per dairy cow and meat per head of cattle are low, the relationship between milk and meat efficiencies is positive. Improved management practices (feed, reproduction, health) increase the feed efficiency of both products. Where milk and meat productivities are high, a trade-off between feed efficiencies of milk and meat can be observed in relation to the share of meat produced in either the dairy sector or the beef sector. As a result, in developing countries, increasing productivities of both dairy and beef cattle herds will increase milk and meat efficiencies, reduce land use and decrease methane emissions. In other regions of the world, increasing meat production from young animals produced by dairy cows is probably a better option to reduce feed use for an unchanged milk-to-meat production ratio.

A 6-month randomized, double-blind, placebo-controlled trial of weekly exenatide in adolescents with obesity.

BACKGROUND: Pharmacological treatment options for adolescents with obesity are very limited. Glucagon-like-peptide-1 (GLP-1) receptor agonist could be a treatment option for adolescent obesity. OBJECTIVE: To investigate the effect of exenatide extended release on body mass index (BMI)-SDS as primary outcome, and glucose metabolism, cardiometabolic risk factors, liver steatosis, and other BMI metrics as secondary outcomes, and its safety and tolerability in adolescents with obesity. METHODS: Six-month, randomized, double-blinded, parallel, placebo-controlled clinical trial in patients (n = 44, 10-18 years, females n = 22) with BMI-SDS > 2.0 or age-adapted-BMI > 30 kg/m2 according to WHO were included. Patients received lifestyle intervention and were randomized to exenatide extended release 2 mg (n = 22) or placebo (n = 22) subcutaneous injections given once weekly. Oral glucose tolerance tests (OGTT) were conducted at the beginning and end of the intervention. RESULTS: Exenatide reduced (P < .05) BMI-SDS (-0.09; -0.18, 0.00), % BMI 95th percentile (-2.9%; -5.4, -0.3), weight (-3 kg; -5.8, -0.1), waist circumference (-3.2 cm; -5.8, -0.7), subcutaneous adipose tissue (-552 cm3 ; -989, -114), 2-hour-glucose during OGTT (-15.3 mg/dL; -27.5, -3.1), total cholesterol (11.6 mg/dL; -21.7, -1.5), and BMI (-0.83 kg/m2 ; -1.68, 0.01) without significant change in liver fat content (-1.36; -3.12, 0.4; P = .06) in comparison to placebo. Safety and tolerability profiles were comparable to placebo with the exception of mild adverse events being more frequent in exenatide-treated patients. CONCLUSIONS: Treatment of adolescents with severe obesity with extended-release exenatide is generally well tolerated and leads to a modest reduction in BMI metrics and improvement in glucose tolerance and cholesterol. The study indicates that the treatment provides additional beneficial effects beyond BMI reduction for the patient group.

Increased lipolysis in LCHAD deficiency.

An increasing number of fatty acid oxidation defects are being detected owing to diagnostic improvements and a greater awareness among clinicians. The metabolic block leads to energy disruption, fatty infiltration, and toxic effects on organ functions exerted by beta-oxidation metabolites. This investigation was undertaken to assess the influence of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency on lipolysis and energy turnover. We addressed the question whether the lipolysis and glucose production rates would be altered in the fasting state in a child with this disease. Lipolysis, glucose production and resting energy expenditure (REE) were studied in a 17-month-old girl with LCHAD deficiency and her healthy twin sister. Lipolysis and glucose production were determined after a 4-6 h fast by constant-rate infusion of [1,1,2,3,3-(2)H(5)]glycerol and [6,6-(2)H(2)]glucose and analysis by gas chromatography-mass spectrometry. REE was estimated by indirect calorimetry. The affected girl showed 50% higher lipolysis than did her sister, whereas the glucose production rates were similar. Plasma levels of dicarboxylic acids of 6-12 carbon atoms chain length, 3-hydroxy fatty acids of 6-18 carbon atoms chain length, total free fatty acids, and acylcarnitines were increased in the patient, as was REE. Since glucose production rates and plasma glucose levels were similar in the two girls, the increased lipolysis observed in the patient probably represents a compensatory mechanism for energy generation. This is achieved at the price of an augmented risk for fatty acid infiltration and toxic effects of beta-oxidation intermediates. This highlights the importance of avoiding fasting in these patients.

Microbiological assessments of compost toilets: in situ measurements and laboratory studies on the survival of fecal microbial indicators using sentinel chambers.

Compost toilet systems were assessed for their ability to reduce microbial indicators and pathogens. Bacterial pathogens were not detected in any samples indicating a low survival rate in composting feces and/or an initial low occurrence. Indicator bacteria showed large variations with no clear trend of lower bacterial numbers after longer storage. In controlled composting experiments, thermophilic conditions were only reached when amendments were made (grass and a sugar solution). Even then it was impossible to ensure a homogenous temperature in the composting fecal material and therefore difficult to achieve a uniform reduction and killing of indicator organisms. Presumptive thermotolerant coliforms, Salmonella typhimurium Phage 28 B and eggs of Ascaridia galli, proved useful as indicators. However, regrowth was detected for enterococci and total numbers of bacteria grown at 36 degrees C. These indicator parameters may therefore overestimate the level of other (pathogenic) bacteria present in the material and can not be recommended for use as reliable indicator organisms in composting toilet systems. The addition of indicator bacteria to fecal material contained in semi-permeable capsules proved to be a useful technique to ensure that microorganisms were contained in a small test volume.

Mutations and allelic loss of p53 in primary tumor DNA from potentially cured patients with colorectal carcinoma.

PURPOSE: To compare p53 alterations in survivors and nonsurvivors after surgery for colorectal cancer. PATIENTS AND METHODS: Twenty-nine potentially cured patients with colorectal carcinoma, without recurrent disease for more than 6 years after their primary surgery, were selected to match a group of 41 colorectal cancer patients with early metastatic spread to the liver. All patients were screened for mutations in the p53 gene, exons 5 to 9, by denaturing gradient gel electrophoresis and subsequent sequencing. RESULTS: The frequency of p53 mutations was significantly different in cured patients (60%) compared with patients with early relapse (41%, P <.05). A significant difference was found in the distribution of mutations, indicating that potentially cured patients had a different proportion of mutations in conserved regions of p53 (P =.02). This difference was explained by a significantly different frequency of mutations in exon 8 (40% v 15%, P =.03), which is part of the conserved region V. All mutations in region V were codon 273 mutations in cured patients, whereas three of four mutations were located in codon 273 in patients with metastatic disease. Allelic loss of p53 (loss of heterozygosity [LOH]) was demonstrated in 26% of the cured patients and in 39% of patients with metastatic disease (P =.36). The combination of mutation and LOH of p53 was the same (17%) in both groups. CONCLUSION: A large number of p53 mutations in colorectal cancer do not promote disease progression. Some mutations, particularly within conserved regions, may even counteract negative functional effects of other p53 structural alterations. A complete loss of p53 function was not related to survival or progression after curative operation of colorectal carcinoma.

Growth hormone-dependent insulin-like growth factor binding protein is a major determinant of bone mineral density in healthy men.

To establish the major determinants of bone mass, we assessed relationships between bone mineral density (BMD) and height, weight, body mass index (BMI), muscle strength, physical capacity (VO2max), body composition, serum concentrations of insulin-like growth factor I (IGF-I), growth hormone (GH), the GH-dependent IGF binding protein (IGFBP-3), testosterone, sex hormone binding globulin (SHBG), osteocalcin, and parathyroid hormone (PTH) in 38 healthy men between 25 and 59 years of age. Values of BMD at all sites (total body, lumbar spine, and hip) were strongly correlated with IGFBP-3 (r = 0.51-0.64, p < 0.001 at all sites), and total-body BMD was also significantly correlated with IGF-I (r = 0.43, p = 0.01). BMD measurements of the total body and of the different sites of the hip were negatively correlated with age and positively with weight, BMI, muscle strength, VO2max, and fat-free weight. IGF-I and IGFBP-3 were both positively related to muscle strength and VO2max. In a stepwise forward multiple-regression analysis, the best model was obtained for the femoral neck, where IGFBP-3, GH, PTH, age, IGF-I, and BMI explained 77% of the variation in BMD. The partial regression coefficients of IGFBP-3, PTH, and BMI were all positive, whereas age, GH, and IGF-I were negatively correlated with BMD. In summary, IGFBP-3 correlated better with BMD than any other study parameter. The findings indicate that GH is of importance for bone mass and suggest that IGFBP-3 not only reflects the integrated GH secretion but also has a direct role in the endocrine regulation of bone metabolism.

Determination of body composition--a comparison of dual-energy x-ray absorptiometry and hydrodensitometry.

Determination of body composition by dual-energy x-ray absorptiometry (DEXA) was evaluated in healthy men, by using underwater weighing (UWW), skinfold thickness measurement, and bioimpedance analysis. There were strong correlations between percent body fat obtained by all techniques, but DEXA gave significantly lower values (P < 0.001). The influence of differences in bone mineral density (BMD) on fat content determined by UWW was also studied. The individual differences between UWW and DEXA fat estimates were calculated and there was a negative correlation with BMD (r = -0.50, P < 0.05). There was also a negative correlation between body fat by UWW and BMD (r = -0.71, P < 0.01) in the subjects with lowest fat by DEXA, indicating that high or low BMD gave false values by UWW. In conclusion, DEXA and UWW provide complementary information and a combination of these techniques seems to offer new opportunities in evaluations of body composition.

Evaluation of modified multicompartment models to calculate body composition in healthy males.

The purpose of this study was to develop flexible and accurate multicompartment equations to calculate body composition and compare the results with methods using common two-compartment equations. Twenty-two healthy male volunteers 22-59 y of age were studied. Body volume was measured by underwater weighing (UWW) or with a skinfold caliper, bone mineral by dual-energy X-ray absorptiometry (DXA), and body water by bioelectrical impedance analysis (BIA). The percentage of water and bone mineral in fat-free mass (FFM) had a significant effect on the difference in percentage fat obtained by the two-compartment model compared with a four-compartment model. FFM density was negatively (r = -0.76, P < 0.001) and percentage water in FFM was positively correlated with age (r = 0.75, P < 0.001). The three-compartment model based on field-adapted methods (skinfold thickness + BIA) to calculate percentage body fat correlated significantly with the more complex four-compartment model (UWW + BIA + DXA; r = 0.95, P < 0.001). The advantages of three- and four-compartment equations are that they compensate for differences in body content of bone mineral and water.

Serum anti-p53 in relation to mutations across the entire translated p53 gene in colorectal carcinomas.

Previous reports have claimed that antibodies to mutated p53 protein indicate poor outcome in malignant disease. The mechanism behind this highly specific process is unclear, although it has been claimed that certain DNA alterations are prone to induction of immune response, since wild-type p53 is almost never immunogenic. The aim of the present analysis was to evaluate whether the presence of anti-p53 was statistically significantly related to any certain DNA alterations in the entirely expressed p53 gene in primary tumors of colorectal cancer. P53 serum antibodies were determined by an enzyme linked immunosorbant assay (ELISA). P53 antibodies were detected in serum of 24 of 88 patients (27%). Twenty-two of 24 (92%) sero-positive patients had mutations in their p53 gene while only 22 of 64 (34%) sero-negative patients had p53 mutations (p<0.01). Mutations were mainly missense with a trend to significantly higher frequency of deletions in sero-negative patients compared to sero-positive subjects (8/25, 32% and 2/22, 9% respectively, p<0.08). Mutations in sero-positive patients were mainly located in exon 5 and 7 and within conserved regions (17 of 22 mutations). In sero-negative patients missense mutations were usually located in exon 5, 7 and 8 being also most frequently located within conserved regions. Most of the p53 deletions in sero-negative patients were however located outside conserved regions (seven of eight deletions). There was no statistical difference between sero-positive and negative patients concerning the spectrum of mutations along the expressed gene. Our study demonstrates that p53 antibodies are usually related to p53 gene mutations but a mutational event is not sufficient to elicit self-immunization. Cellular protein binding to p53 or individual differences of major histocompatibility complex based presentation of p53 protein sequences by immune cells is therefore the most likely explanation between sero-negative and sero-positive patients.

Dissemination of vancomycin-resistant enterococci harboring vanA through disposal of waste derived from industrial production of vancomycin.

We demonstrated the occurrence of vancomycin-resistant enterococci (VRE) in waste derived from the industrial production of vancomycin and their dissemination through disposal of such waste into a sewage treatment plant. Bacteriological counts on a medium selective for enterococci (Slanetz-Bartley agar) revealed the presence of high numbers of presumptive VRE (approximately 10(6) CFU/ml) in the waste originating from the fermentation biomass used for vancomycin production. The waste was also found to contain active residues of vancomycin (64-1,024 microg/ml) by bioassays using a vancomycin-susceptible enterococcal strain. Randomly amplified polymorphic DNA (RAPD) analysis of 65 presumptive VRE isolates from the waste allowed distinction of four genotypes, two of which (A and D) belonged to the genus Enterococcus, most likely E. faecium, and harbored the vanA gene conferring high-level vancomycin resistance. The same VRE strains found in the waste occurred also in the biological tanks and the final effluent of the sewage treatment plant receiving the waste, as demonstrated by the detection of undistinguishable pulsed-field gel electrophoresis (PFGE) patterns in VRE isolated from these sources. These results indicate the need to assess the possible dissemination of VRE and other antibiotic-resistant bacteria through disposal of waste derived from antibiotic production.

The influence of physical activity on BMR.

In addition to factors such as fat free mass, hormonal status, genetics and energy balance, previous physical activity has been shown to influence energy turnover during resting (RMR = resting metabolic rate) or basal conditions (BMR = basal metabolic rate). This article presents data on BMR from elite endurance athletes (4 female and 4 male), at least 39 h after their last training session, in comparison with sedentary nonathletic controls matched for sex and fat free mass (FFM). Comparisons with theoretical calculations of BMR were also made. The athletes were shown to have a significantly higher BMR than was expected from calculations based on body mass (16%, P < 0.05) or body composition (12%, P < 0.05). There were no corresponding differences found in the nonathletic control group. The athletes had a 13% higher (P < 0.001) BMR than controls if related to FFM and 16% (P = 0.001) if related to both FFM and fat mass (FM). The athletes were also found to have 10% lower R-values (P < 0.01) indicating higher fat oxidation. The conformity of these findings with the present literature and the possible mechanisms behind them as well as its influence on theoretical calculations of energy turnover (ET) based on activity factors expressed as multiples of RMR are further discussed.

Evaluation of modified multicompartment models to calculate body composition in healthy males.

Body volume was measured by underwater weighing (UWW) or with a skinfold caliper; bone mineral by dual energy X-ray absorptiometry (DXA); and body water by bioelectrical impedance analysis (BIA) in 22 healthy males. The percentage of water and bone mineral in fat-free mass had a significant effect on the calculated amount of fat using a two-compartment model. A three-compartment model based on field-adapted methods (skinfold thickness + BIA) to calculate body fat, correlated significantly with a more complex four-compartment model (UWW + BIA + DXA) (r = 0.95, p < 0.001). The advantages of three- and four-compartment equations are that they reduce the number of assumptions.

Escherichia coli contamination of lettuce grown in soils amended with animal slurry.

A pilot study was conducted to assess the transfer of Escherichia coli from animal slurry fertilizer to lettuce, with E. coli serving as an indicator of fecal contamination and as an indicator for potential bacterial enteric pathogens. Animal slurry was applied as fertilizer to three Danish agricultural fields prior to the planting of lettuce seedlings. At harvest, leaves (25 g) of 10 lettuce heads were pooled into one sample unit (n = 147). Soil samples (100 g) were collected from one field before slurry application and four times during the growth period (n = 75). E. coli was enumerated in slurry, soil, and lettuce on 3M Petrifilm Select E. coli Count Plates containing 16 mg/liter streptomycin, 16 mg/liter ampicillin, or no antimicrobial agent. Selected E. coli isolates (n = 83) originating from the slurry, soil, and lettuce were genotyped by pulsed-field gel electrophoresis (PFGE) to determine the similarity of isolates. The slurry applied to the fields contained 3.0 to 4.5 log CFU/g E. coli. E. coli was found in 36 to 54% of the lettuce samples, streptomycin-resistant E. coli was found in 10.0 to 18.0% of the lettuce samples, and ampicillin-resistant E. coli in 0 to 2.0% of the lettuce samples (the detection limit was 1 log CFU/g). The concentration of E. coli exceeded 2 log CFU/g in 19.0% of the lettuce samples. No E. coli was detected in the soil before the slurry was applied, but after, E. coli was present until the last sampling day (harvest), when 10 of 15 soil samples contained E. coli. A relatively higher frequency of E. coli in lettuce compared with the soil samples at harvest suggests environmental sources of fecal contamination, e.g., wildlife. The higher frequency was supported by the finding of 21 different PFGE types among the E. coli isolates, with only a few common PFGE types between slurry, soil, and lettuce. The frequent finding of fecal-contaminated lettuce indicates that human pathogens such as Salmonella and Campylobacter can be present and represent food safety hazards.

Isotope dilution ESI-LC-MS/MS for quantification of free and total Nε-(1-Carboxymethyl)-L-Lysine and free Nε-(1-Carboxyethyl)-L-Lysine: comparison of total Nε-(1-Carboxymethyl)-L-Lysine levels measured with new method to ELISA assay in gruel samples.

ESI-LC-MS/MS method with isotope dilution and SPE based on cation-exchange was developed for determination of free and total Nε-(1-Carboxymethyl)-L-Lysine (CML) and free Nε-(1-Carboxyethyl)-L-Lysine (CEL). The use of nonafluoropentanoic acid in mobile phase was omitted, SPE recoveries of 82±3% and 91±10% (n=6) for CML and CEL respectively and, calibration curves (R(2)>0.9985) were attained. The method was applied to gruel samples and LoQ for the method was 5 ng/ml, RSD <10% and accuracy was 115%. Total CML levels in the gruel samples varied from 103-408 mg/kg protein. Free CML levels which were 1000 times lower than total CML were three times higher than free CEL levels. CML in a gruel sample was 127±7, 84±9 and 253±28 mg/kg using the current ESI-LC-MS/MS, ELISA and GC-MS respectively. The described method has advantages over ELISA with respect to reproducibility and specificity and over GC-MS with respect to reproducibility.

Physical capacity, respiratory quotient and energy expenditure during exercise in male patients with schizophrenia compared with healthy controls.

BACKGROUND: Despite massive research on weight gain and metabolic complications in schizophrenia there are few studies on energy expenditure and no current data on physical capacity. AIM: To determine oxygen uptake capacity, respiratory quotient (RQ) and energy expenditure during a submaximal exercise test in patients with schizophrenia and healthy controls. METHOD: Ten male patients and 10 controls were included. RQ and energy expenditure were investigated with indirect calorimetry during a cycle ergometer test. The submaximal work level was defined by heart rate and perceived exhaustion. Physical capacity was determined from predicted maximal oxygen uptake capacity (VO(2-max)). RESULTS: The patients exhibited significantly higher RQ on submaximal workloads and lower physical capacity. A significant lower calculated VO(2-max) remained after correction for body weight and fat free mass (FFM). Energy expenditure did not differ on fixed workloads. CONCLUSION: RQ was rapidly increasing in the patients during exercise indicating a faster transition to carbohydrate oxidation and anaerobic metabolism that also implies a performance closer to maximal oxygen uptake even at submaximal loads. This may restrict the capacity for everyday activity and exercise and thus contribute to the risk for weight gain. Physical capacity was consequently significantly lower in the patients.

Escherichia coli contamination and health aspects of soil and tomatoes (Solanum lycopersicum L.) subsurface drip irrigated with on-site treated domestic wastewater.

Faecal contamination of soil and tomatoes irrigated by sprinkler as well as surface and subsurface drip irrigation with treated domestic wastewater were compared in 2007 and 2008 at experimental sites in Crete and Italy. Wastewater was treated by Membrane Bio Reactor (MBR) technology, gravel filtration or UV-treatment before used for irrigation. Irrigation water, soil and tomato samples were collected during two cropping seasons and enumerated for the faecal indicator bacterium Escherichia coli and helminth eggs. The study found elevated levels of E. coli in irrigation water (mean: Italy 1753 cell forming unit (cfu) per 100 ml and Crete 488 cfu per 100 ml) and low concentrations of E. coli in soil (mean: Italy 95 cfu g(-1) and Crete 33 cfu g(-1)). Only two out of 84 tomato samples in Crete contained E. coli (mean: 2700 cfu g(-1)) while tomatoes from Italy were free of E. coli. No helminth eggs were found in the irrigation water or on the tomatoes from Crete. Two tomato samples out of 36 from Italy were contaminated by helminth eggs (mean: 0.18 eggs g(-1)) and had been irrigated with treated wastewater and tap water, respectively. Pulsed Field Gel Electrophoresis DNA fingerprints of E. coli collected during 2008 showed no identical pattern between water and soil isolates which indicates contribution from other environmental sources with E. coli, e.g. wildlife. A quantitative microbial risk assessment (QMRA) model with Monte Carlo simulations adopted by the World Health Organization (WHO) found the use of tap water and treated wastewater to be associated with risks that exceed permissible limits as proposed by the WHO (1.0 × 10(-3) disease risk per person per year) for the accidental ingestion of irrigated soil by farmers (Crete: 0.67 pppy and Italy: 1.0 pppy). The QMRA found that the consumption of tomatoes in Italy was deemed to be safe while permissible limits were exceeded in Crete (1.0 pppy). Overall the quality of tomatoes was safe for human consumption since the disease risk found on Crete was based on only two contaminated tomato samples. It is a fundamental limitation of the WHO QMRA model that it is not based on actual pathogen numbers, but rather on numbers of E. coli converted to estimated pathogen numbers, since it is widely accepted that there is poor correlation between E. coli and viral and parasite pathogens. Our findings also stress the importance of the external environment, typically wildlife, as sources of faecal contamination.