Diversity and relatedness of Shiga toxin-producing Escherichia coli and Campylobacter jejuni between farms in a dairy catchment.

The aim of this study was to examine the population structure, transmission and spatial relationship between genotypes of Shiga toxin-producing Escherichia coli (STEC) and Campylobacter jejuni, on 20 dairy farms in a defined catchment. Pooled faecal samples (n = 72) obtained from 288 calves were analysed by real-time polymerase chain reaction (rtPCR) for E. coli serotypes O26, O103, O111, O145 and O157. The number of samples positive for E. coli O26 (30/72) was high compared to E. coli O103 (7/72), O145 (3/72), O157 (2/72) and O111 (0/72). Eighteen E. coli O26 and 53 C. jejuni isolates were recovered from samples by bacterial culture. E. coli O26 and C. jejuni isolates were genotyped using pulsed-field gel electrophoresis and multilocus sequence typing, respectively. All E. coli O26 isolates could be divided into four clusters and the results indicated that E. coli O26 isolates recovered from calves on the same farm were more similar than isolates recovered from different farms in the catchment. There were 11 different sequence types of C. jejuni isolated from the cattle and 22 from water. An analysis of the population structure of C. jejuni isolated from cattle provided evidence of clustering of genotypes within farms, and among groups of farms separated by road boundaries.

Effect of using a stand-off pad on Campylobacter jejuni strain diversity in a herd of dairy cows.

AIM: To determine the effect of stand-off pad (SOP) use on the prevalence and strain diversity of Campylobacter jejuni in a small herd of dairy cows. METHODS AND RESULTS: Faecal samples were collected from 21 cows on four sampling occasions (events), one in each season, over 1 year. The cows usually grazed on pasture but during winter they spent 18 h a day on a SOP. Campylobacter prevalence ranged from 48-52% on pasture but was 62% on the SOP. The diversity of 386 C. jejuni isolates was determined using Enterobacterial Repetitive Intergenic Consensus polymerase chain reaction (ERIC/PCR). There were 11 ERIC types identified for the herd over the course of the study. Of those 11, four to seven (per event) were present when the cows were grazing pasture but only two during SOP use. CONCLUSIONS: The use of the SOP was associated with an increase in prevalence and a reduction in diversity of C. jejuni. SIGNIFICANCE AND IMPACT OF THE STUDY: The reduction in ERIC types on the SOP indicated an increase in transfer of only some strains of C. jejuni among the cows. One of these strains persisted throughout the study. The zoonotic potential of this strain warrants further investigation.

Campylobacter jejuni inactivation in New Zealand soils.

AIM: The study was undertaken to determine the inactivation rate of Campylobacter jejuni in New Zealand soils. METHODS AND RESULTS: Farm dairy effluent (FDE) inoculated at c. 10(5) ml(-1) with C. jejuni was applied to intact soil cores at a rate of 2 l m(-2). Four soils were used: Hamilton (granular); Taupo (pumice); Horotiu and Waihou (allophanic). After FDE application cores were incubated at 10 degrees C for up to 32 days. For all four soils all the FDE remained within the cores and at least 99% of C. jejuni were retained in the top 5 cm. Campylobacter jejuni had declined to the limit of detection (two C. jejuni 100 g(-1)) by 25 days in Hamilton and Taupo soils and by 32 days in Waihou soil. In contrast, in Horotiu soil the decline was only three orders of magnitude after 32 days. Simulated heavy rainfall was applied 4 and 11 days after FDE application and only about 1% of the applied C. jejuni were recovered in leachates. CONCLUSIONS: This study demonstrated that at least 99% of applied C. jejuni were retained in the top 5 cm of four soils where they survived for at least 25 days at 10 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: Soil retention of C. jejuni is efficient at FDE application rates that prevent drainage losses. The low infectious dose of C. jejuni and its ability to survive up to 25 days have implications for stock management on dairy farms.

Effect of plant growth on dehydration rates and microbial populations in sewage biosolids.

The high water content of sewage biosolids makes them expensive to transport. Two experiments were done to see if it was practical to use transpiration by plants as a low cost method to dehydrate biosolids. (i) To assess biosolids as a growth-medium for plants, maize, beans, pumpkin, forage oats and an annual ryegrass were grown in pots of fresh biosolids. Plant growth varied between species and potassium deficiency was found to be a limiting factor for barley. Roots were slow to penetrate anoxic biosolids in the bottom of the pots. (ii) Dehydration rates were measured in 30 litre boxes of biosolids from two different sources. Boxes were planted with maize or beans, or kept fallow. Despite the high (73-83%) initial water content of the biosolids, plants were susceptible to wilting on warm days which suggested that a significant proportion of the water in biosolids is not freely available to plants. Shrinkage caused a major reduction in biosolids volume in both experiments. When change in volume of the biosolids residue was accounted for, there was a 56% average water loss in planted treatments and 34% water loss in the fallow treatment. This indicates that planting may have some potential as a technique to accelerate dehydration of biosolids. Water contents were not reduced sufficiently to influence biosolids microbial populations.

Faecal bacteria yields in artificial flood events: quantifying in-stream stores.

Stream sediments have been recognised as an in-channel store of faecal contamination that can be mobilised during floods or other sediment-disturbing events. We studied this store of faecal contamination by creating artificial floods during dry weather when, in the absence of overland flow from the catchment, the only source of faecal bacteria was stores within the channel. Artificial floods, created by releasing water from a supply reservoir, increased the E. coli concentration in the water column by two orders of magnitude, from a background level of 10(2) cfu per 100 mL to over 10(4) cfu per 100 mL. The bacterial peak concentrations and yields declined systematically through a triplicate flood series. The size of the total in-channel store, calculated as the sum of yields of an infinite series of artificial floods, was approximately 10(8) cfu m(-2) of streambed area. Direct measurements of sediment E. coli found few sites (only those associated with cattle crossings) with areal concentrations as high as 10(8) cfu m(-2), consistent with flood yields. Concentrations of E. coli in the biofilms on exposed rocks were orders of magnitude lower, indicating that exposed rocks were not a source of E. coli released by the artificial floods.

Faecal contamination over flood events in a pastoral agricultural stream in New Zealand.

Faecal bacterial dynamics during flood events were studied in the Topehaehae Stream near Morrinsville, New Zealand, in a catchment used for grazing dairy and beef cattle. During the rising limb of a natural flood event, E. coli bacterial concentration rose by more than 2 orders of magnitude and peaked at 41,000 cfu/100 mL. E. coli correlated closely with turbidity over the flood event, and both variables peaked close to the time of maximum flow acceleration rather than peak flow. An artificial flood on the same stream, created by releasing water from a supply reservoir during fine weather with no wash-in from the catchment, produced a broadly similar pattern of faecal contamination (peak E. coli = 12,500 cfu/100 mL). This and other evidence suggests that direct deposition of faecal matter by cattle in the stream channel may be of similar or greater importance than wash-in from land. The flood experiments have been useful for constructing a model of faecal bacterial yields, and they imply that exclusion of livestock from stream channels may appreciably improve water quality.

Description of Caldicellulosiruptor saccharolyticus gen. nov., sp. nov: an obligately anaerobic, extremely thermophilic, cellulolytic bacterium.

A new obligately anaerobic, extremely thermophilic, cellulolytic bacterium is described. The strain designated Tp8T 6331 is differentiated from thermophilic cellulolytic clostridia on the basis of physiological characteristics and phylogenetic position within the Bacillus/Clostridium subphylum of the Gram-positive bacteria. Strain Tp8T 6331 is assigned to a new genus Caldicellulosiruptor, as Caldicellulosiruptor saccharolyticus gen., nov., sp. nov.

Sunlight inactivation of enterococci and fecal coliforms in sewage effluent diluted in seawater.

Inactivation (loss of culturability) by sunlight of enterococci and fecal coliforms within sewage effluent diluted in seawater was investigated in field experiments. In most experiments, 500-ml flasks of pure silica were used to confine activated sludge effluent diluted to 2% (vol/vol) in seawater. Inactivation of bacteria in these flasks (diameter, 0.1 m) was faster than in either open chambers (depth, 0.25 m) or patches of dyed effluent (depth of order, 1 m), probably because of the longer light paths in the latter two types of experiment, which caused greater attenuation of sunlight. Inactivation of 90% of enterococci generally required 2.3 times the insolation required for 90% inactivation of fecal coliforms, because of both the presence of larger initial shoulders on survival curves and a lower final inactivation rate. Two parameters are required to model inactivation of enterococci, a shoulder constant as well as a rate coefficient. The depth dependence of inactivation rate for both fecal indicators matched the attenuation profile of UV-A radiation at about 360 nm. Inactivation by UV-B radiation (290 to 320 nm), which penetrates much less into seawater, is of minor importance compared with the UV-A and visible radiation in sunlight, contrary to expectations in consideration of published action spectra for bacterial inactivation.

Quality control of bacterial enumeration.

Standard bacterial suspensions can be used to assess test method performance, via control charts, and inhibition of recovery when analyzing water samples. Variability in standard suspensions prepared from different strains and species and the use of frozen environmental samples for quality control for spore and bacteriophage analyses are also discussed.

The degradation of lignocellulosics by extremely thermophilic microorganisms.

Five extremely thermophilic cellulose-degrading isolates obtained from New Zealand thermal springs were tested for their ability to degrade a number of natural lignocellulosic substrates. Degradation by three of the isolates was generally similar to that by the moderate thermophile, Clostridium thermocellum but occurred at a higher temperature. The New Zealand isolates were also found to grow on xylan as sole carbohydrate source, which probably extends their attack to hemicellulose.