Growth of Dehalococcoides strains with chlorophenols as electron acceptors

Dehalococcoides strains reductively dechlorinate a wide variety of halogenated compounds including chlorinated benzenes, biphenyls, naphthalenes, dioxins, and ethenes. Recent genome sequencing of the two Dehalococcoides strains CBDB1 and 195 revealed the presence of 32 and 18 reductive dehalogenase homologous genes, respectively, and therefore suggested an even higher dechlorinating potential than previously anticipated. Here, we demonstrate reductive dehalogenation of chlorophenol congeners by Dehalococcoides strains CBDB1 and 195. Strain CBDB1 completely converted 2,3-dichlorophenol, all six trichlorophenols, all three tetrachlorophenols, and pentachlorophenol to lower chlorinated phenols. Observed dechlorination rates in batch cultures with cell numbers of 10(7) mL(-1) amounted up to 35 microM day(-1). Chlorophenols were preferentially dechlorinated in the ortho position, but also doubly flanked and singly flanked meta- or para-chlorine substituents were removed. We used a newly designed computer-assisted direct cell counting protocol and quantitative PCR to demonstrate that strain CBDB1 uses chlorophenols as electron acceptors for respiratory growth. The growth yield of strain CBDB1 with 2,3-dichlorophenol was 7.6 x 10(13) cells per mol of Cl- released, and the growth rate was 0.41 day(-1). For strain 195, fast ortho dechlorination of 2,3-dichlorophenol, 2,3,4-trichlorophenol, and 2,3,6-trichlorophenol was detected, with only the ortho chlorine removed. Because chlorinated phenolic compounds are widely distributed as natural components in anaerobic environments, our results reveal one mode in which the Dehalococcoides species could have survived through earth history.     

Stressful life events and smoking behaviors in Chinese adolescents: a longitudinal analysis.

The associations between stressful life events and smoking have been established among adolescents in the United States. However, whether these relationships are similar in adolescents from other non-Western cultures is unknown. Understanding these relationships in adolescents may help to provide opportunities to reduce the smoking rates in those cultures by providing positive coping methods that do not include smoking. In this longitudinal study, the associations between nine stressful life events scales and smoking behaviors were examined in a sample of Chinese adolescents. Six of these scales, positive school-related, negative school-related, positive family-related, positive peer-related, negative peer-related, and negative health-related had significantly different means among females and males. Among males, positive school-related stress was a protective factor for smoking susceptibility. Among females, positive school-related stress was a protective factor and negative school-related stress was a risk factor for lifetime smoking, and negative family-related stress was a risk factor for smoking susceptibility. Findings indicate that smoking among male adolescents in China may not be the result of stress; however, in females stress may contribute to the decision to smoke. Future directions are discussed.     

Quantitative determination of fluorotelomer sulfonates in groundwater by LC MS/MS

Aqueous film-forming foams (AFFF) are complex mixtures containing fluorocarbon- and hydrocarbon-based surfactants that are used to fight hydrocarbon-fueled fires. The military is the largest consumer of AFFF in the United States, and fire-training activities conducted at military bases have led to groundwater contamination by unspent fuels and AFFF chemicals. A direct-injection, liquid-chromatography tandem mass spectrometry (LC MS/MS) method was developed to quantify a suite of fluorotelomer sulfonate surfactants in groundwater collected from military bases where fire-training activities were conducted. The 4:2, 6:2, and 8:2 fluorotelomer sulfonates were detected and quantified in groundwater from two of the three military bases. The total fluorotelomer sulfonate concentrations observed at Wurtsmith AFB, MI, and Tyndall AFB, FL, ranged respectively from below quantitation (< or = 0.60) to 182 microg/L and from 1100 to 14,600 microg/L. Analyses of a fluorotelomer-based AFFF concentrate by negative ion fast atom bombardment/mass spectrometry and LC MS/MS analyses indicate that the AFFF concentrate contains only a small amount of fluorotelomer sulfonates and that fluoroalkylthioamido sulfonates are the main anionic fluorosurfactant in the mixtures. More research is needed to determine the fate of fluoroalkylthioamido sulfonates in the environment.     

Sampling strategies for assessing lameness,injuries, and body condition score on dairy farms

Our objective was to evaluate how sampling strategies (i.e., how many cows to sample and which animals to include) used in 4 dairy cattle welfare assessment programs affect the classification of dairy farms relative to thresholds of acceptability for animal-based measures. We predicted that classification performance would improve when more cows were sampled and when selecting from all lactating cows versus when some pens were excluded. On 38 freestall farms, we assessed all 12,375 cows for lameness, injuries on the tarsal (hock) and carpal joints, and body condition score and calculated the farm-level prevalence for each measure. Based on approaches used in the industry, we evaluated 6 sampling strategies generated using formulas with precision (d) of 15, 10, or 5% applied to either a single high-producing pen or all lactating cows; an additional sample was included with d = 10% applied to the entire herd, selecting lactating cows in proportion to their representation in the herd. For each sampling strategy, cow records were selected randomly (in 10,000 replicates) to calculate prevalence. The strategy of assessing all cows in the high-producing pen was also compared. Farms were classified as meeting (below) or failing to meet (above) thresholds of ≤15% moderate lameness; ≤20% moderate carpal or hock injuries; <10, <5, and ≤1% severe lameness; or injuries on the carpus or hock; and <5, <3, <1, or 0% thin cows. For each measure and threshold, we calculated pooled percent agreement, kappa, sensitivity, specificity, and positive and negative predictive value for each sampling strategy using true prevalence as the gold standard for herd classification. Across measures and thresholds, classification performance increased with the number of cows sampled [i.e., when narrower precision values (d = 5 vs. 10 vs. 15%) were used in the sample size calculation]. Because narrower precision values can dramatically increase sample size, assessment programs may need to consider both feasibility and the degree of misclassification they will accept. Applying the formula directly to lactating cows performed better than applying it to the entire herd and then selecting lactating cows in proportion to their representation in the herd. Farm classifications were similar whether cows in the hospital pen were included or excluded from the sample. Selecting all cows from the high-producing pen resulted in classifications similar to when including all lactating cows, suggesting that assessing cows from the high-producing pen may serve as an acceptable proxy for all lactating cows on the farm.     

Introduction of sulphydryl groups into pea vicilin: Formation of intra-and inter-polypeptide disulphide bonds

Site directed mutagenesis was used to introduce two cysteine residues into the hydrophilic regions of the pea vichlin polypeptide previously expressed in Saccharomyces cerevisiae. The mutated polypeptide was expresed and the resultant protein was characterised by sodium dodecyl sulphatepolyacrylamide gel electrophoresis. The presence of both intra- and inter-polypeptide sulphydryl bonds was indicated by bands of different mobility from those of the native polypeptide. The association of mutated vicili polypeptides into disulphide-bonded aggregates within the yeast was not a random process, trimers being the major aggregate produced. When the mutated vicilin was extracted from yeast and purified, random aggregates of the vicilin polypeptide were formed.     

Volatile compounds in Bacillus‐fermented soybeans

Thua nao, a traditional, proteolytic, fermented soybean condiment of northern Thailand, was prepared from cooked whole soybeans by natural flora fermentation. The microbial flora during the fermentation was dominated by Bacillus species. The formation of volatile compounds during the fermentation was studied. In addition, the volatile compounds of two samples of commercial dried thua nao and two samples of commercial Japanese natto were analysed. Fermentation led to a large increase in the concentration of total volatile compounds, from 35 µg kg^?1 wet weight in cooked soybeans to 3500 µg kg^?1 wet weight in 72 h fermented material. The major volatile compounds in fermented beans were 3‐hydroxybutanone (acetoin), 2‐methlybutanoic acid, pyrazines, dimethyl disulphide and 2‐pentylfuran. Sun drying of 72 h fermented material resulted in the loss of 65% of total volatiles, including important aroma compounds. The commercial dried thua nao samples had low concentrations of total volatile compounds (380 µg kg^?1 wet weight). It is suggested that improved drying/preservation methods are needed to retain aroma compounds in the traditional products. The natto samples were devoid of aldehydes, aliphatic acids and esters, and sulphur compounds, whereas the thua nao samples contained a diversity of these compounds. Previous investigators have reported these compounds in natto and it is not possible to suggest the existence of systematic differences between the volatile compounds in traditional thua nao prepared with an undefined, mixed microbial flora and those in natto fermented with Bacillus subtilis. © 2001 Society of Chemical Industry     

Effect of Heat Processing on IgE Reactivity and Cross‐Reactivity of Tropomyosin and Other Allergens of Asia‐Pacific Mollusc Species: Identification of Novel Sydney Rock Oyster Tropomyosin Sac g 1

1 Scope

Shellfish allergy is an increasing global health priority, frequently affecting adults. Molluscs are an important shellfish group causing food allergy but knowledge of their allergens and cross‐reactivity is limited. Optimal diagnosis of mollusc allergy enabling accurate advice on food avoidance is difficult. Allergens of four frequently ingested Asia‐Pacific molluscs are characterized: Sydney rock oyster (Saccostrea glomerata), blue mussel (Mytilus edulis), saucer scallop (Amusium balloti), and southern calamari (Sepioteuthis australis), examining cross‐reactivity between species and with blue swimmer crab tropomyosin, Por p 1.

2 Methods and results

IgE ELISA showed that cooking increased IgE reactivity of mollusc extracts and basophil activation confirmed biologically relevant IgE reactivity. Immunoblotting demonstrated strong IgE reactivity of several proteins including one corresponding to heat‐stable tropomyosin in all species (37–40 kDa). IgE‐reactive Sydney rock oyster proteins were identified by mass spectrometry, and the novel major oyster tropomyosin allergen was cloned, sequenced, and designated Sac g 1 by the IUIS. Oyster extracts showed highest IgE cross‐reactivity with other molluscs, while mussel cross‐reactivity was weakest. Inhibition immunoblotting demonstrated high cross‐reactivity between tropomyosins of mollusc and crustacean species.

3 Conclusion

These findings inform novel approaches for reliable diagnosis and improved management of mollusc allergy.     

Outdoor grazing of dairy cows on pasture versus indoor feeding on total mixed ration: Effects on gross composition and mineral content of milk during lactation

The influence of feeding system and lactation period on the gross composition, macroelements (Ca, P, Mg, and Na), and trace elements (Zn, Fe, Cu, Mo, Mn, Se, and Co) of bovine milk was investigated. The feeding systems included outdoor grazing on perennial ryegrass pasture (GRO), outdoor grazing on perennial ryegrass and white clover pasture (GRC), and indoors offered total mixed ration (TMR). Sixty spring-calving Holstein Friesian dairy cows were assigned to 3 herds, each consisting of 20 cows, and balanced with respect to parity, calving date, and pre-experimental milk yield and milk solids yield. The herds were allocated to 1 of the 3 feeding systems from February to November. Milk samples were collected on 10 occasions over the period June 17 to November 26, at 2 or 3 weekly intervals, when cows were on average 119 to 281 d in lactation (DIL). The total lactation period was arbitrarily sub-divided into 2 lactation periods based on DIL, namely mid lactation, June 17 to September 9 when cows were 119 to 203 DIL; and late lactation, September 22 to November 26 when cows were 216 to 281 DIL. With the exception of Mg, Na, Fe, Mo, and Co, all other variables were affected by feeding system. The GRO milk had the highest mean concentrations of total solids, total protein, casein, Ca, and P. The TMR milk had the highest concentrations of lactose, Cu, and Se, and lowest level of total protein. The GRC milk had levels of lactose, Zn, and Cu similar to those of GRO milk, and concentrations of TS, Ca, and P similar to those of TMR milk. Lactation period affected all variables, apart from the concentrations of Fe, Cu, Mn, and Se. On average, the proportion (%) of total Ca, P, Zn, Mn, or Se that sedimented with the casein on high-speed ultracentrifugation at 100,000 × g was ≥60%, whereas that of Na, Mg, or Mo was ≤45% total. The results demonstrate how the gross composition and elemental composition of milk can be affected by different feeding systems.