Biomagnification of alpha- and gamma-hexabromocyclododecane isomers in a Lake Ontario food web

The extent of bioaccumulation of hexabromocyclododecane (HBCD) isomers (alpha, beta, and gamma) was determined in the Lake Ontario pelagic food web using liquid chromatography tandem mass spectrometry (LC/MS/MS). Concentrations of the alpha-isomer were consistently higher than that of the gamma-isomer. The beta-isomer was below method detection limits in all samples. Whole body concentrations (ng/g, wet wt) of alpha- and gamma-HBCD were highest in the top predator lake trout samples ranging from 0.4 to 3.8 ng/g for the alpha-isomer and 0.1 to 0.8 ng/g for the gamma-isomer. For the prey fish species, the trends in alpha- and gamma-HBCD levels were slimy sculpin > smelt > alewife. Mean concentrations of total (sigma) HBCD (sum of alpha- and gamma-isomers) in the macrozooplankter Mysis relicta (0.14 +/- 0.02 ng/g wet wt) and in the benthic invertebrate Diporeia hoyi (0.16 +/- 0.02 ng/g, wet wt) were similar and approximately twice as high as in plankton (0.06 +/- 0.02 ng/g, wet wt). A strong positive linear relationship was found between sigmaHBCD concentrations (wet wt) and trophic level based on delta15N suggesting that HBCD biomagnifies in the Lake Ontario food web. The trophic magnification factor (TMF = 6.3) derived from the slope of the sigmaHBCD - trophic level relationship was slightly higher than TMFs for p,p'-DDE (6.1) and sigmaPCBs (5.7) found previously. Biomagnification factors (BMF, calculated as the ratio of lipid corrected concentration in predator/lipid corrected concentration in prey) were variable between feeding relationships and ranged from 0.4 to 10.8 for the alpha-isomer and from 0.2 to 10 for gamma-isomers.     

Characterization of O157:H7 and other Escherichia coli isolates recovered from cattle hides, feces, and carcasses

In a previous study, the seasonal prevalence was reported for stx+ Escherichia coli O157:H7 in feces and on hides and carcasses of cattle at processing. Overall, 1,697 O157:H7 isolates have now been characterized for the incidence of (i) eae(O157), hlyA, stx1, and stx2 in the recovered isolates and (ii) presumptive rough and presumptive nonmotile isolates. Seven O157:H7 isolates (0.4%) lacked stx genes, although they carried eae and hlyA. All but one of the isolates carried both eae and hlyA. Approximately two-thirds of the isolates (64% when one isolate per sample was considered) carried both stx1 and stx2. E. coli O157:H7 cells that harbored both stx1 and stx2 were more often recovered from hides in the fall (79% of the fall hide isolates) and winter (84% of the winter hide isolates) than in the spring (53%) and summer (59%). Isolates recovered from preevisceration carcasses showed a similar but not statistically significant trend. Twenty-three of the 25 O157:H7 isolates carrying stx1 but not stx2 were recovered during summer. Fifteen presumptive rough and 117 presumptive nonmotile stx+ O157:H7 isolates were recovered. Ten (67%) of the presumptive rough isolates were recovered during summer. Ninety-five of the presumptive nonmotile isolates (81%) were recovered during fall. Forty-eight percent of the false-positive isolates (175 of 363) tentatively identified as O157:H7 were O157+ H7- and lacked eae(O157), hlyA, and stx. These data suggest that in beef processing samples (i) there are minor seasonal variations in the prevalence of stx genes among E. coli O157:H7 isolates, (ii) presumptive rough and presumptive nonmotile stx+ O157:H7 isolates are present, (iii) E. coli O157:H7 isolates lacking stx genes may be rare, and (iv) O157+ H7- isolates lacking stx genes can result in many false-positive results.     

Isolation of an acetyl-CoA synthetase gene (ZbACS2) from Zygosaccharomyces bailii

A gene homologous to Saccharomyces cerevisiae ACS genes, coding for acetyl-CoA synthetase, has been cloned from the yeast Zygosaccharomyces bailii ISA 1307, by using reverse genetic approaches. A probe obtained by PCR amplification from Z. bailii DNA, using primers derived from two conserved regions of yeast ACS proteins, RIGAIHSVVF (ScAcs1p; 210-219) and RVDDVVNVSG (ScAcs1p; 574-583), was used for screening a Z. bailii genomic library. Nine clones with partially overlapping inserts were isolated. The sequenced DNA fragment contains a complete ORF of 2027 bp (ZbACS2) and the deduced polypeptide shares significant homologies with the products of ACS2 genes from S. cerevisiae and Kluyveromyces lactis (81% and 82% identity and 84% and 89% similarity, respectively). Phylogenetic analysis shows that the sequence of Zbacs2 is more closely related to the sequences from Acs2 than to those from Acs1 proteins. Moreover, this analysis revealed that the gene duplication producing Acs1 and Acs2 proteins has occurred in the common ancestor of S. cerevisiae, K. lactis, Candida albicans, C. glabrata and Debaryomyces hansenii lineages. Additionally, the cloned gene allowed growth of S. cerevisiae Scacs2 null mutant, in medium containing glucose as the only carbon and energy source, indicating that it encodes a functional acetyl-CoA synthetase. Also, S. cerevisiae cells expressing ZbACS2 have a shorter lag time, in medium containing glucose (2%, w/v) plus acetic acid (0.1-0.35%, v/v). No differences in cell response to acetic acid stress were detected both by specific growth and death rates. The mode of regulation of ZbACS2 appears to be different from ScACS2 and KlACS2, being subject to repression by a glucose pulse in acetic acid-grown cells.     

Wet deposition of polychlorinated biphenyls in the eastern Mediterranean

The concentrations of polychlorinated biphenyls (PCBs) were measured in rain samples collected from a semiurban and a marine background site of the eastern Mediterranean. The concentration of sigmaPCB (sum of 54 PCB congeners) in the city of Heraklion (2.9 +/- 1.9 ng L(-1)) was not significantly higher than the corresponding concentration measured at the background sampling station of Finokalia (1.9 +/- 0.9 ng L(-1)). In both sites, the sum of tri- and tetrachlorinated congeners accounted for more than 55% of sigmaPCB in rainwater. For all samples, the percentage of particle-bound PCBs ranged between 6.6% and 63.8%, providing an average value of 31 +/- 18%. The washout ratios of particulate PCBs (WP) were constant for individual congeners regardless the degree of chlorination. Average WP values ranged between 1.9 x 10(5) and 5.2 x 10(5) while a value of 2.7(+/- 1.3) x 10(5) was deduced for sigmaPCB. The corresponding washout ratios for gaseous PCBs were substantially lower and ranged between 7 x 10(3) (PCB 99) and 1.3 x 10(5) (PCB 180). Washout ratios of gaseous PCBs were also calculated based on Henry's law, and they were found to be 30-920 times lower than those obtained from field measurements. On the basis of our data, the wet deposition flux of sigmaPCB in the eastern Mediterranean should approach 820 ng m(-2) yr(-1). This flux is similar with the values recently reported for several background sites of the United States and Europe, but it is 1 order of magnitude lower than the flux of PCBs measured in the western Mediterranean 16 yr ago.     

Inactivation of pathogenic bacteria by cucumber volatiles (E,Z)-2,6-nonadienal and (E)-2-nonenal

The effects of (E,Z)-2,6-nonadienal (NDE) and (E)-2-nonenal (NE) on Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium were investigated. A suspension of each organism of 6 to 9 log CFU/ml was incubated for 1 h at 37 degrees C in brain heart infusion solution that contained 0 to 500 or 1,000 ppm of NDE or NE. Depending on concentration, exposure to either NDE or NE caused a reduction in CFU of each organism. Treatment with 250 and 500 ppm NDE completely eliminated viable B. cereus and Salmonella Typhimurium cells, respectively. L. monocytogenes was the most resistant to NDE, showing only about a 2-log reduction from exposure to 500 ppm for 1 h. Conversely, this concentration of NDE caused a 5.8-log reduction in E. coli O157:H7 cells. NE was also effective in inactivating organisms listed above. A higher concentration of NE, 1,000 ppm, was required to kill E. coli O157:H7, L. monocytogenes, or Salmonella Typhimurium compared with NDE. In conclusion, both NDE and NE demonstrated an apparent bactericidal activity against these pathogens.     

Hexavalent uranium diffusion into soils from concentrated acidic and alkaline solutions

Uranium contamination of soils and sediments often originates from acidic or alkaline waste sources, with diffusion being a major transport mechanism. Measurements of U(VI) diffusion from initially pH 2 and pH 11 solutions into a slightly alkaline Altamont soil and a neutral Oak Ridge soil were obtained through monitoring uptake from boundary reservoirs and from U concentration profiles within soil columns. The soils provided pH buffering, resulting in diffusion at nearly constant pH. Micro X-ray absorption near edge structure spectra confirmed that U remained in U(VI) forms in all soils. Time trends of U(VI) depletion from reservoirs and U(VI) concentration profiles within soil columns yielded Kdvalues consistent with those determined in batch tests at similar concentrations (approximately 1 mM) and much lower than values for sorption at much lower concentrations (nM to microM). These results show that U(VI) transport at high concentrations can be relatively fast at non-neutral pH, with negligible surface diffusion, because of weak sorption.     

High levels of the mitochondrial large ribosomal subunit protein 40 prevent loss of mitochondrial DNA in null mmf1 Saccharomyces cerevisiae cells

Members of the YERO57c/YJGFc/UK114 protein family have been identified in bacteria and eukaryotes. The budding yeast Saccharomyces cerevisiae contains two different proteins of this family, Hmf1p and Mmf1p. We have previously shown that Mmf1p is a mitochondrial protein functionally related to its human homologue and able to influence the maintenance of mitochondrial DNA. Deletion of Mmf1 results in loss of the mitochondrial genome. Using a multicopy suppression approach, we have identified a protein of the mitochondrial large ribosomal subunit, MRPL40, which stabilizes mtDNA in Deltammf1 cells. Overexpression of MRPL40 did not prevent loss of mtDNA in a mutant strain lacking the mitochondrial protein Abf2p. Thus, MRPL40 does not have a general effect on mtDNA stability, but it may be specific for the mmf1-null strain. We also show that the Deltamrpl40 cells present a similar phenotype to the mmf1-null strain, having reduced mtDNA stability and growth rate. Furthermore, we observed that rho(+)Deltamrpl40 haploid cells can be obtained when tetrads are directly dissected on medium containing a non-fermentable carbon source. Thus, replication and segregation of the mtDNA can occur in the absence of MRPL40. We also show that another mitochondrial ribosomal protein, MRPL38, is able to overcome the Deltammf1-associated defect. Together, our results suggest a link between Mmf1p and the two mitochondrial ribosomal proteins.     

Impact of intervention strategies on Listeria contamination patterns in crawfish processing plants: a longitudinal study

Two ready-to-eat crawfish processing plants were monitored for 2 years to study the impact of Listeria control strategies, including employee training and targeted sanitation procedures, on Listeria contamination. Environmental, raw material, and finished product samples were collected weekly during the main processing months (April to June) and tested for Listeria spp. and Listeria monocytogenes. Before implementation of control strategies (year 1), the two processing plants showed Listeria spp. prevalences of 29.5% (n = 78) in raw, whole crawfish, 5.2% (n = 155) in the processing plant environment, and 0% (n = 78) in finished products. In year 2, after plant-specific Listeria control strategies were implemented, Listeria spp. prevalence increased in raw crawfish (57.5%, n = 101), in the processing plant environment (10.8%, n = 204), and in the finished product (1.0%, n = 102). Statistical analysis showed a significant increase in Listeria spp. prevalence (P < 0.0001) and a borderline nonsignificant increase in L. monocytogenes prevalence (P = 0.097) on raw material in year 2. Borderline nonsignificant increases were also observed for Listeria spp. prevalence in environmental samples (P = 0.082). Our data showed that Listeria spp. prevalence in raw crawfish can vary significantly among seasons. However, the increased contamination prevalence for raw materials only resulted in a limited Listeria prevalence increase for the processing plant environment with extremely low levels of finished product contamination. Heat treatment of raw materials combined with Listeria control strategies to prevent cross-contamination thus appears to be effective in achieving low levels of finished product contamination, even with Listeria spp. prevalences for raw crawfish of more than 50%.     

Systematic clinical examinations for identification of latent udder health types in Danish dairy herds

A cross-sectional study was conducted to explore the applicability of systematic clinical examinations of udders as an additional tool for the evaluation of udder health status on dairy farms. During 2000, each of the 16 dairy farms was visited 5 times; 20 cows per farm were chosen at random at each visit for clinical udder examination immediately after milking. The clinical examination included both pathological and morphological variables. One examination per cow was included in the analysis (n = 707 cows). Principal component analysis (PCA) was performed in 3 steps. First, 19 variables characterizing udder and teats were analyzed (PCA 1). Second, the variables parity and stage of lactation were included (PCA 2). Finally, somatic cell count (SCC) and milk yield (PCA 3) were included. The PCA resulted in 4 components that explained 30% of the variation of the data: 1) small udder, 2) distressed udder, 3) mastitis udder, and 4) soiled udder. Variables with high positive correlation to the "small udder" were small udder shape, short teats, and first parity. Impaired teat surface, hard udder texture, and a long udder shape were related to the "distressed udder." The "mastitis udder" was characterized by the clinical variables asymmetry between front quarters, asymmetry between hind quarters, knotty tissue, and acute clinical mastitis. Reduced milk yield and high SCC were related to the "mastitis udder," whereas low SCC was related to the "small udder." The "soiled udder" was related to early lactation. Including this information in the assessment of udder health may be of substantial value for data analysis in farms with suspected under-reporting of clinical mastitis.