Reports of honey bee population decline has spurred many national efforts to understand the extent of the problem and to identify causative or associated factors. However, our collective understanding of the factors has been hampered by a lack of joined up trans-national effort. Moreover, the impacts of beekeeper knowledge and beekeeping management practices have often been overlooked, despite honey bees being a managed pollinator. Here, we established a standardised active monitoring network for 5 798 apiaries over two consecutive years to quantify honey bee colony mortality across 17 European countries. Our data demonstrate that overwinter losses ranged between 2% and 32%, and that high summer losses were likely to follow high winter losses. Multivariate Poisson regression models revealed that hobbyist beekeepers with small apiaries and little experience in beekeeping had double the winter mortality rate when compared to professional beekeepers. Furthermore, honey bees kept by professional beekeepers never showed signs of disease, unlike apiaries from hobbyist beekeepers that had symptoms of bacterial infection and heavy Varroa infestation. Our data highlight beekeeper background and apicultural practices as major drivers of honey bee colony losses. The benefits of conducting trans-national monitoring schemes and improving beekeeper training are discussed.

Little has been published about the occurrence, speciesidentification, andpathogenicpotentialofcoagulasenegative staphylococci (CoNS) present in drinking water. In this study, ten species were identified among 57 isolates of staphylococci from 756 samples of chlorinated drinking water taken from public distribution networks in the Slovak Republic. S. warneri (37%), S. haemolyticus (23%), and S. saprophyticus ssp. saprophyticus (14%) were identified most frequently. Isolates did not produce coagulase, DNase, or hyaluronidase; production of gelatinase and lecithinase was observed in 28 and 22 isolates, respectively. Genetically encoded ability for production of enterotoxin SED was revealed in two isolates. Among ten antibiotics tested, resistance to ampicillin (66.7%), penicillin (64.9%), and erythromycin (57.9%) were observed most frequently. Resistance to gentamicin, vancomycin, or clindamycin was not confirmed. Production of β-lactamase was observed in 64.9% of isolates. Fourty-two isolates were resistant to two or more antibiotics tested, and eight isolates showed multiresistance. The presence of mecA gene was confirmed in 8 isolates, while PBP2a was revealed in 7. Two isolates of S. epidermidis were identified as methicillin-resistant (MRSE). The results demonstrate that CoNS in chlorinated drinking water may possess virulence factors and show resistance to various antibiotics. Therefore, their pathogenic potential should not be ignored.

Epidemiological studies have emphasized the importance of Enterococcus spp. in regard to positive and negative role of these bacteria. Presented work investigated a relationship among some factors (cytolysin, gelatinase, esp and biofilm formation) on phenotypic and genotypic level in relation to antibiotic resistance (gentamicin -GEN, streptomycin-STR, and vancomycin-VAN) in 82 enterococcal isolates (18 isolates of Enteroccus faecium, 26 of E. faecalis, 35 of E. casseliflavus and 3 of E. gallinarum) taken from cloacal swabs of poultry and from the water intended for livestock breeding. For phenotypic determination, commercial cultivation methods were used. Genes for appropriate factors were detected by polymerase chain reaction. Among tested isolates, 53 (65%) carried gelE gene and consequently 43 (52%) isolates produced gelatinase. The presence of cyl operon was detected in 53 (65%) and production of haemolysin on horse blood agar was determined in 37 (45%). In 14 (17%) isolates, esp gene was detected, but 28 (34%) isolates including those without esp gene detection were considered as strong biofilm producers. In 11 isolates (13%), all three genes (esp/gelE/cylA) were present contrary to 18 (22%) strains with negative PCR reaction. Resistance to GEN and STR was detected in 1 and 3 isolates respectively. All enterococci tested were susceptible to VAN. No relationship between virulence factors and resistance to antimicrobials was found. The data of this study proved relatively wide distribution of virulence factors among enterococci of animal and environmental origin and suggested that such isolates can be a reservoir of virulence trait.

An occurrence of resistance to tetracycline (TET) and erythromycin (ERY) was ascertained in 82 isolates of Enterococcus spp. of animal and environmental origin. Using E test, 33 isolates were resistant to TET and three isolates to ERY. Using polymerase chain reaction (PCR; single and multiplex), the TET determinants tet(M) and tet(L) were detected in 35 and 13 isolates, respectively. Twelve isolates carried both tet(M) and tet(L) genes. Eight isolates possessed ermB gene associated with ERY resistance. Multiplex PCR was shown to be a suitable method for simultaneous determination of all three resistance determinants that occurred most frequently in bacteria isolated from poultry. This study also demonstrates that gastrointestinal tract of broilers may be a reservoir of enterococci with acquired resistance to both TET and ERY that can be transferred to humans via food chain.

The purpose of this study was to evaluate the discriminatory power of ddl-PCR (D-alanin-D-alanin ligase PCR) and ITS-PCR (Internal Transcribed Spacer PCR) for accurate identification of enterococcal species in comparison with phenotypic assays. Results confirm previous published data that ddl-PCR simple approach allows rapid identification of two the most frequently isolated spp. E. faecalis and E. faecium. When identification points towards other enterococci then that mentioned above, the ITS-PCR seems to be suitable complementary assay. Correct identification of enterococci to species level is important because of different spp. susceptibility to some clinically important antibiotics. For example, it is necessary to distinquish acquired vancomycin resistance from inherent one that is less epidemiologically important. Additionally, both methods can be valuable in epidemiological studies following enterococcus resistance gene transfer within human population.

The role of susceptibility or resistance to antimicrobial agents in enterococci is important for the food industry. Eighty two isolates (18 Enterococcus faecium, 26 E. faecalis, 35 E. casseliflavus and 3 E. gallinarum) were taken from cloacal swabs of poultry and from the water intended for livestock breeding during the years 2004-2006. The isolates were evaluated for susceptibility to antimicrobial agents: ampicillin, gentamicin, streptomycin, erythromycin, tetracycline, and vancomycin determined by the disk diffusion test and E-test. All isolates were found to be susceptible to ampicillin and vancomycin. Resistance rates to gentamicin, streptomycin, tetracycline and erythromycin were 1%, 4%, 40% and 4%, respectively. One isolate of E. faecium was resistant to gentamicin and erythromycin. Two and six isolates of this species were resistant to streptomycin and tetracycline, respectively. One isolate of E. faecalis was resistant to streptomycin and erythromycin. High level resistance to tetracycline was observed in E. faecalis (46%) and E. casseliflavus (37%) isolates. One isolate of E. casseliflavus was resistant to erythromycin. E. gallinarum isolates showed a low level of resistance to gentamicin, streptomycin, tetracycline and erythromycin (0-2%). This research suggests that enterococci of animal and environmental origin may play a potential role as a reservoir of resistance to antimicrobial agents.

A long-term sufficient intake of fruits and vegetables reduces significantly the risk of gastric and colorectal carcinoma. It is anticipated that natural antioxidants are involved in this effect in addition to other substances. The aim of this study was to determine levels of vitamins A, C and E, as well as beta-carotene, selenium, zinc and copper in blood of 249 patients with precancerous lesions (atrophic gastritis, gastric hyperplastic polyp, gastric, colonic and rectal adenoma, chronic ulcerative colitis) and in 96 individuals with gastric, colonic or rectal carcinoma and to compare these levels with the values of a control group of 130 healthy individuals. We have found that the frequency of average values of analyzed micronutrients in precancerous groups was decreasing in the order vit C > vit E/vit A > Se > beta-car. The average levels of vitamins and beta-carotene were significantly reduced in all carcinoma groups, while selenium level showed a decrease only in the gastric carcinoma group. Copper level was elevated in the ulcerative colitis group and in all groups with carcinoma. The results indicate a frequent insufficient saturation of organism by natural antioxidants in groups with precancerous lesions and carcinomas of stomach and colorectum. Therefore, it is necessary to increase the general consumption of fruits and vegetables in Slovakia as a part of primary prevention of malignant diseases in these organs. Chemoprevention may be recommended in individuals with precancerous lesions.

BACKGROUND:In comparison to food habits of non-vegetarians alternative nutrition bears several risks, but it also has beneficial effects. Considering risks represent a significant age aspect. Nutritional risk and protective factors were evaluated in a adolescent group.METHODS AND RESULTS:Food habits of 53 lactoovovegetarians (mean duration of vegetarianism 3.2 y) assessed by analysis of dietary questionnaires were compared to those of 55 nonvegetarians and evaluated with respect to basic biochemical indicators (iron, hemoglobin, calcium, total protein, cholesterol, fatty acids, vitamin A, beta-carotene, vitamin C, vitamin 3). Vegetarians consume less fat, and they have higher intake of plant fat, which is reflected din more favourable lipid levels, i.e. lower risk parameters of atherosclerosis (cholesterol, LDL-cholesterol, atherogenic index, saturated fatty acids) and higher values of factors with antisclerotic effect (32.9% ratio of HDL-cholesterol vs. 30.6% in non-vegetarians, polyunsaturated fatty acids, vitamin E/cholesterol ratio, vitamin C). Total plasma protein levels are balanced in both groups (intake of full value mixture of amino acids from grains, legumes (soya), and mild in vegetarians, similar intake of total proteins with a small difference in plant protein consumption). Iron and calcium absorption is inhibited in vegetarians as a consequence of presence of plant food--physic acid, oxaloacetic acid (20.7% hyposiderinemia vs. 0% in non-vegetarians, high iron intake in both groups, iron from plant sources represented 82% in vegetarian boys and 95% in vegetarian girls, hypocalcemia in 28% of vegetarians vs. 7.3% of non-vegetarians). Vegetarians have significantly higher levels of all antioxidant vitamins. This is result of higher consumption of vegetables, fruit, plant fat and sprouts. Higher intake and higher levels of antioxidants are reflected in significantly lower lipoperoxidation values.CONCLUSIONS:Lactoovovegetarian nutrition represents risk for adolescents if the values of iron and calcium are considered, however it has positive aspects on lipid parameters, protein values, and antioxidant parameters.

Risk nutritional factors of alternative alimentation detected in childhood were evaluated in a group of adult lactoovovegetarians (n = 47). The levels of iron, calcium, zinc, total proteins, gluthatione, plasmatic profile of fatty acids and the lipoperoxidation product in correlation with the values of antioxidative vitamins were studied. The results are compared with mean sample upon mixed nutrition (omnivores n = 42). In both groups were the mean values of iron, calcium and zinc in physiological range, but significantly lower in vegetarians. In the alternative nutrition group was in 21 percent of probands hyposiderinemia detected (vs 5 percent in the omnivores group), in 19 percent of probands hypocalcemia (vs 9 percent) and in 6 percent hypozincemia (vs 0 percent). Full-bodied mixture of milk proteins, egg proteins and vegetable sources in lactoovovegetarians ensured sufficient protein saturation and caused significantly higher level of blood gluthatione (intake also in food). Increased value of fatty acid peroxidation index was not due to increased lipoperoxidation in lactoovovegetarians--significantly lower levels of conjugated fatty acid dienes were determined. This was ensured by sufficient protection by means of essential antioxidants--the levels of vitamins E, C, beta-carotene are in vegetarians significantly higher. These are overtreshold values representing reduced risk of free-radical diseases. Lactoovovegetarians had significantly higher content of linoleic and alpha-linoleic acids in plasma. Values of polyunsaturated fatty acids with C20 and C22 and 3-6 double bonds were similar to values in omnivores. In probands on alternative nutrition with iron deficit was significantly lower activity of delta 6 desaturase determined. (Tab. 3, Ref. 45.)

The aim of the present study was to assess serum levels of copper and zinc levels and erythrocytes Cu,Zn-SOD activity and to determine probable changes in gastric and colorectal precancerous diseases, benign breast diseases, gastric, colorectal and breast cancer. The study included 165 subjects with cancer, 348 subjects with precancerous (atrophic gastritis, gastric adenoma, colon adenoma, rectal adenoma) and/or benign diseases (weak dysplasia, severe dysplasia, fibroadenoma, cystic disease) and 161 randomly selected healthy controls. Our results suggest that while in gastric and colorectal cancer there were mostly increased copper levels, in breast cancer they were not changed. Zinc levels were weakly decreased in atrophic gastritis, gastric adenoma and breast cancer. There was a strong positive correlation between zinc levels and SOD activity in fibroadenoma and a weak positive correlation in colorectal adenoma and colorectal cancer without any correlation between SOD activity and copper in these groups. In gastric precancerous disease there was a positive correlation between SOD and copper. The results of this study suggest that serum trace element levels and activity of related enzymes might be different in various neoplastic processes. This variation in neoplastic processes might be influenced by other factors that have to be considered in complex relationships between the whole body and neoplastic cells.

BACKGROUND:Low protein quality and quantity is reported to be a possible risk of alternative nutrition. Pulses contain 18-41% of methionine in relation to reference protein, moreover, its content in cereals is by one half lower. Therefore vegetarians and vegans may have an insufficient intake of sulphur-containing amino acids that may subsequently affect glutathione values (precursors of its synthesis).METHODS AND RESULTS:In groups of adults on an alternative diet--lactoovovegetarians (n = 47) and vegans (n = 44) aged 19-62 years with average duration on a vegetarian or vegan diet of 7.6 and 4.9 years, respectively, glutathione levels (GSH) were measured in erythrocytes (spectrophotometrically), as well as the activity of GSH-dependent enzymes. As nutritional control (n = 42) served an average sample of omnivores selected from a group of 489 examined, apparently healthy subjects of the same age range living in the same region. One to low protein intake (56% of RDA) exclusively of plant origin significantly lower levels of total proteins were observed in vegans with a 16% frequency of hypoproteinaemia (vs 0% in omnivores). In comparison to omnivores a significantly lower glutathione level was found (4.28 +/- 0.12 vs 4.84 +/- 0.14 mumol/g Hb, P < 0.01). Lactoovovegetarians because of their protein intake in adequate amounts with a 27% proportion of animal proteins (dairy products, eggs) consume a balanced mixture of amino acids, which is reflected in total protein levels similar to omnivores and significantly higher values of glutathione -5.26 +/- 0.12 mumol/g Hb, P < 0.05 (intake of glutathione in diet, higher consumption of fruit and vegetable in comparison to omnivores). A sufficient supply of glutathione as the substrate for enzymatic reactions of hydrogen peroxide or lipid hydroperoxide catabolism, as well as for detoxication of xenobiotics, was reflected in lactoovovegetarians in a significantly higher activity of glutathione-peroxidase and glutathione-S-transferase in erythrocytes.CONCLUSIONS:Low protein intake exclusively of plant origin, significantly lower protein levels with 16% frequency of hypoproteinaemia, significantly lower glutathione values in blood in comparison to omnivores and lactoovovegetarians confirm the risk of a vegan diet also in adult age.

Blood serum selenium levels were measured in 891 healthy children and adolescents (aged 11–18 yr, 450 girls and 441 boys) residing in both rural and urban areas from eight regions of Slovakia. Subjects were divided into four age groups (11–12 y, 13–14 y, 15–16 y, and 17–18 y). Serum selenium concentration was determined by the electrothermal atomic absorption spectrometric method. The mean (±SD) serum selenium concentrations were 0.750 ±0.255 µmol/L in girls and 0.773 ±0.235 µmol/L in boys. A large proportion of the individuals (25.7% in girls, 18.1% in boys) exhibited serum selenium levels under 0.57 µmol/L (45 µg/L). An increasing trend of the serum selenium values with age has been observed in both boys ( p < 0.01) and girls ( p < 0.05). Boys had higher serum selenium levels in the all age groups but the differences were not statistically significant.