Rapid Gas Chromatographic Technique for Presumptive Detection of Clostridium botulinum in Contaminated Food

A simple gas-liquid chromatography end-product assay is reported for butyric and other short-chain fatty acids as presumptive indicators of Clostridium botulinum contamination in food.     

Assessing Climate Variability Impact on Thermotolerant Coliform Bacteria in Surface Water

This study investigated the impacts of climate variability on thermotolerant coliform bacteria (TCB) transport in the Upper Pearl River watershed (UPRW) in Mississippi. The Soil and Water Assessment Tool (SWAT) was applied using daily observed stream flows and TCB concentration data. The SWAT model was successfully calibrated and validated using both manual and automatic methods from February 2011 to June 2012 (NSE and R² up to 0.79). The Long Ashton Research Station Weather Generator (LARS-WG), a stochastic weather generator, with the global climate model, CCSM3, which was developed by the U.S. National Center for Atmospheric Research (NCAR) was used for future climate variability simulations. The Special Report on Emissions Scenarios (SRES) A1B of the Intergovernmental Panel on Climate Change (IPCC) was simulated for the mid (2046–2065) and late (2080–2099) 21st century. The SWAT model simulated TCB concentrations in surface water and demonstrated reasonable performances (R² up to 0.59 and NSE up to 0.58). During mid-century climate, average monthly TCB levels increase to 175%, while late-century average monthly TCB levels increase to 297% from the watershed. Although late-century climate variability impacts were determined more critical than mid-century climate impacts, appropriate watershed management practices are required to adapt to maintain and improve water quality.     

Internal Standards for Gas Chromatographic Analysis of Metabolic End Products from Anaerobic Bacteria

2-Methylpentanoic acid and benzoic acid are suggested for use as routine internal standards for gas chromatographic analysis of microbial end products.     

Numerical Taxonomy Analysis of Bacteria Isolated from the Completed 'Most Probable Numbers' Test for Coliform Bacilli

Lactose-fermenting bacteria were isolated from oyster, water and sediment samples tested to determine the Most Probable Number (MPN) of coliforms present. The completed test series was used to enumerate the total number of coliforms. The strains from the completed tests of the MPN analysis were compared with named reference cultures, for which 100 phenotypic characters were recorded and the taxonomic data analysed by methods of numerical taxonomy. The isolates were recovered in 13 phenetic groups, defined at or above the 80% similarity level. Nine phena were identified as Acinetobacter calcoaceticus, Chromobacterium violaceum, Citrobacter freundii, Erwinia herbicola, Escherichia coli, Hafnia alvei, Klebsiella pneumoniae, Serratia liquefaciens , and Ser. marcescens. Of these, Ac. calcoaceticus and C. violaceum were isolated exclusively from oyster tissue samples and strains of the other taxa from both water and oyster samples. Some strains of Esch. coli, Er. herbicola, H. alvei and bacteria that were not Enterobacteriaceae were found to lose the ability to ferment lactose, following storage on laboratory media. Identifications from diagnostic keys, rapid identification systems and IMViC patterns were not always in agreement.     

Coliform Bacteria as Indicators of Diarrheal Risk in Household Drinking Water: Systematic Review and Meta-Analysis

Background

Current guidelines recommend the use of Escherichia coli (EC) or thermotolerant (“fecal”) coliforms (FC) as indicators of fecal contamination in drinking water. Despite their broad use as measures of water quality, there remains limited evidence for an association between EC or FC and diarrheal illness: a previous review found no evidence for a link between diarrhea and these indicators in household drinking water.

Objectives

We conducted a systematic review and meta-analysis to update the results of the previous review with newly available evidence, to explore differences between EC and FC indicators, and to assess the quality of available evidence.

Methods

We searched major databases using broad terms for household water quality and diarrhea. We extracted study characteristics and relative risks (RR) from relevant studies. We pooled RRs using random effects models with inverse variance weighting, and used standard methods to evaluate heterogeneity and publication bias.

Results

We identified 20 relevant studies; 14 studies provided extractable results for meta-analysis. When combining all studies, we found no association between EC or FC and diarrhea (RR 1.26 [95% CI: 0.98, 1.63]). When analyzing EC and FC separately, we found evidence for an association between diarrhea and EC (RR: 1.54 [95% CI: 1.37, 1.74]) but not FC (RR: 1.07 [95% CI: 0.79, 1.45]). Across all studies, we identified several elements of study design and reporting (e.g., timing of outcome and exposure measurement, accounting for correlated outcomes) that could be improved upon in future studies that evaluate the association between drinking water contamination and health.

Conclusions

Our findings, based on a review of the published literature, suggest that these two coliform groups have different associations with diarrhea in household drinking water. Our results support the use of EC as a fecal indicator in household drinking water.     

Direct Test for Altered Gas Exchange Rates in Water stressed Soybean Nodules

Water stress usually lowers the nitrogenase activity of soybeanroot nodules. This loss in activity might result from an increasedbarrier to nodular gas exchange, from a general reduction inbiochemical function, or both. To test for the possibility ofan increased barrier to gas diffusion, we measured the apparentlag time for initiation of acetylene reduction by intact soybeanplants, both before and after water stress. Mild nodular waterloss (i.e. 10% of fresh weight or less) seldom altered the apparentlag time, whereas severe water stress (20–40% f. wt loss)frequently produced a small increase in apparent lag time. Severewater stress also produced a large decrease (24%) in the externaldiameter of the nodules and a loss of the white lenticel traces.Water stress usually caused a decrease in the apparent K_m foracetylene. The data do not suggest a large change in the diffusiveresistance to acetylene of nodules subjected to water stress.Thus, the observed decrease in nitrogenase activity may resultprimarily from biochemical, rather than physical, changes. However,because of the relatively greater importance of gas-phase diffusionfor oxygen entry, we cannot exclude the possibility of a largechange in a small gas pathway that affects oxygen influx morethan acetylene influx. Diffusion, Glycine max, nitrogen fixation, water stress     

Liquid Chromatographic Procedure for Fermentation Product Analysis in the Identification of Anaerobic Bacteria

High-performance liquid chromatography with a cation-exchange resin-packed column was used to determine fermentation products of several known and unknown Clostridium species. The column was operated at 30°C, and isocratic elution was done with 0.013 N H₂SO₄. Sample preparation for high-performance liquid chromatographic analysis required only membrane filtration. Glucose and formate were readily determined. Quantitative results were easily obtained. Chromatograms of eight unknown strains could be matched with chromatograms of at least one of the type culture strain chromatograms. In some cases, additional testing was necessary before identification could be made. The same conclusions were reached by parallel testing with gas chromatography to determine fermentation products. High-performance liquid chromatography is simple to apply and, under some conditions, is faster than gas chromatography for fermentation product analysis.     

Presence and Sources of Fecal Coliform Bacteria in Epilithic Periphyton Communities of Lake Superior

Epilithic periphyton communities were sampled at three sites on the Minnesota shoreline of Lake Superior from June 2004 to August 2005 to determine if fecal coliforms and Escherichia coli were present throughout the ice-free season. Fecal coliform densities increased up to 4 orders of magnitude in early summer, reached peaks of up to 1.4 × 10⁵ CFU cm⁻² by late July, and decreased during autumn. Horizontal, fluorophore-enhanced repetitive-PCR DNA fingerprint analyses indicated that the source for 2% to 44% of the E. coli bacteria isolated from these periphyton communities could be identified when compared with a library of E. coli fingerprints from animal hosts and sewage. Waterfowl were the major source (68 to 99%) of periphyton E. coli strains that could be identified. Several periphyton E. coli isolates were genotypically identical (≥92% similarity), repeatedly isolated over time, and unidentified when compared to the source library, suggesting that these strains were naturalized members of periphyton communities. If the unidentified E. coli strains from periphyton were added to the known source library, then 57% to 81% of E. coli strains from overlying waters could be identified, with waterfowl (15 to 67%), periphyton (6 to 28%), and sewage effluent (8 to 28%) being the major potential sources. Inoculated E. coli rapidly colonized natural periphyton in laboratory microcosms and persisted for several weeks, and some cells were released to the overlying water. Our results indicate that E. coli from periphyton released into waterways confounds the use of this bacterium as a reliable indicator of recent fecal pollution.