Effectiveness of a steam-vacuum sanitizer for reducing Escherichia coli O157:H7 inoculated to beef carcass surface tissue

A steam-vacuum sanitizer reduced aerobic plate counts associated with bovine faecal contamination from 5.5 log₁₀ cfu cm⁻² to 3.0 ± 0.21 log₁₀ cfu cm⁻² on beef carcass short plates. The same beef carcass short plates inoculated wiht 7.6 ± 0.09 log₁₀ cfu cm⁻² Escherichia coli O157: H7 in faeces, yielded an average residual level of E. coli O157: H7 of 2.1 ± 0.21 log₁₀ cfu cm⁻² after steam-vacuum treatments. This study demonstrates the effectiveness of a steam-vacuum sanitizer for removing E. coli O157: H7 from beef carcasses.     

Routine metabolism of juvenile spot, Leiostomus xanthums (Lacépède), as a function of temperature, salinity and weight

Routine oxygen consumption rates of juvenile spot, Leiostomus xanthums , were measured over a range of temperatures, salinities and fish weights. As predicted, Q O₂ increased with temperature and decreased with body weight. However, Q O₂ decreased with decreasing salinity and did not show the expected minimum at isosmotic concentrations. The data are best described by the relationship: log₁₀ Q O₂ (mg O₂ g⁻¹ h⁻¹) = 0.129 log_lo salinity (%0) + 1.604 log₁₀ temperature (°C)-0.1401og₁₀(g)-2.767.     

A desk study of the relationship between temperature and hatching time for the eggs of five species of salmonid fishes

SUMMARY. Information on temperature (T°C) and time from fertilization to 50% hatch ( D days) for five species of salmonid fishes has been used to assess several mathematical models relating D and T . No single equation gave the best fit to all five data sets. The power law with temperature correction (α), log₁₀₁ D = log₁₀ a + b log₁₀ ( T - α) and the quadratic, log₁₀ D = log₁₀ a + bT + b ₁ T ² (where a, b, b ₁, and α are constants), each accounted for over 97 % of the variance of D and were good fits to the observed data points for all five species. There was little difference between the predictions obtained from these two equations within the range of observed temperatures. Therefore, the simpler power-law model is preferred. However, there were substantial within-species differences between values of D predicted from extrapolations of the two models from 2 or 3°C down to 0°C. When more data for low temperatures become available it will be possible to make a more objective choice of model.     

Note: Thermophilic campylobacters in dairy slurries on Lancashire farms: seasonal effects of storage and land application

Slurry storage tanks on Lancashire farms were sampled in May, June, October and November. There was a seasonal pattern in the number of thermophilic campylobacters. The average log₁₀ MPN per gram fresh weight (log₁₀ MPN gfw⁻¹) in stored samples in May and June was 0·78 ( S.D. 0·71) compared to 2·07 ( S.D. 0·70) in November and December. Campylobacters were readily detected in samples of mature slurry and composted bedding that farmers were about to put to land. The survival of campylobacters in slurry sprayed on land was studied in two seasons. In June, on farm A, stored slurry was mechanically aerated for 48 h and very low numbers of campylobacters (0·9 log₁₀ MPN gfw⁻¹) remained in the slurry before it was sprayed onto land. They became undetectable within 24 h once sprayed on land. In contrast, campylobacters in matured but unaerated slurry that was sprayed onto land on farm B in February were still detectable in samples taken 5 d after application to land, dropping from 2·11 log₁₀ MPN gfw⁻¹–1·37, a decline of only 0·74 log₁₀ MPN gfw⁻¹ in the first 5 d after application.     

The Effect of Temperature, Pressure and Chlorine Concentration of Spray Washing Water on Numbers of Bacteria on Lamb Carcases

Combined analysis of three experiments showed that when lamb carcases with initial bacterial numbers of between logi₁₀3.29 and 4.22/cm² were spray washed, statistically significant reductions in bacterial numbers of log₁₀O.5 were obtained when the spray wash water temperature was > 57°C, and reductions of log₁₀1.0 were obtained when the temperature was ≥ 80°C. Reductions at all temperatures were enhanced by log₁₀0.66 when the water contained 30 µg/ml chlorine, but increasing the concentration to 450 µg/ml reduced bacterial numbers only by a further log₁₀0–29. At highly contaminated sites increasing the duration of spraying from 30 to 120 s significantly increased the reductions obtained when water containing added chlorine was used. Reductions in bacterial numbers after spray washing with pressures of 3.5, 5.6. 7.7 kg/cm² were not significantly different.     

The effect of spray washing on the development of bacterial numbers and storage life of lamb carcases

Total bacterial numbers at the most highly contaminated sites on lamb carcases (in the crutch region and adjacent to the abdominal incision) were significantly reduced to log₁₀ 3.3/cm² when spray washed with unchlorinated water at 80°C and to log₁₀ 2.8 when water at 80°C containing 450 μg/ml chlorine was used, whereas numbers on carcases which were cloth cleaned or spray washed with water at 10°C remained at approximately log₁₀ 4.0. During refrigerated storage, however, carcases treated by all methods developed similar numbers of bacteria and had the same storage life, evidently because spray washing did not affect numbers of bacteria on the diaphragm. Although initial numbers of bacteria at this site were low (log₁₀ 2.9), their numbers, and also the amount of slime, increased more rapidly there than at other sites. In addition, spray washing did not significantly affect numbers of Pseudomonas spp or Brochothrix thermosphacta , which accounted for <1% of the microflora after slaughter at each site but whose numbers were between log₁₀ 6.1 and 7.5/cm² when carcases were rejected as spoiled.     

Population variability among three small mammal species in the semiarid Neotropics: the role of density-dependent and density-independent factors

We addressed the role of density-dependent (direct and delayed) and density-independent (precipitation) factors in shaping the dynamics of fluctuating populations of three small mammal species. Using a stepwise regression procedure, we tested the effects of nonlagged population density (log₁₀ N_t-1), lagged population density (log₁₀ N_t-2), and annual precipitation on the per capita rate of population change of Phyllotis darwini, Akodon olivaceus , and Thylamys elegans in two habitat types of a semiarid region of Chile. The most irruptive species ( P. darwini ) showed direct and delayed density-dependent effects in equatorial subpopulation, and only direct density-dependence in polar subpopulation. The per capita rates of population change of A. olivaceus showed direct density-dependent and precipitation effects in both habitats types, while T. elegans showed direct density-dependence and precipitation effects in the equatorial subpopulation but only a marginal effect of direct density-dependence in the polar subpopulation. The presence of delayed density-dependent strongly suggests the importance of biological interactions in shaping the dramatic irruptions exhibited by P. darwini.     

The use of agarose wedge-gel electrophoresis for resolving both small and large naturally occurring plasmids

A wedge-shaped, horizontal agarose gel gave better electrophoretic resolution of both large and small plasmids than conventional linear gels. The wedge-gel increased mobility and separation of the larger plasmids whilst retarding the mobility of the smaller plasmids and tightening the bands. The linearity of the relationship between log₁₀ molecular size and log₁₀ relative mobility, for a range of plasmids from 2.1 to 221 kb, was increased. Therefore, estimations of plasmid sizes are more accurate using a wedge-gel than with conventional gels.     

Survival of Escherichia coli O157 : H7 in yoghurt during preparation and storage at 4°C

Cow's milk was inoculated with ca 10³ and 10⁷ cfu ml⁻¹ Escherichia coli O157 : H7. After fermentation at 42°C for 0–5 h, the yoghurt was stored at 4°C. Two kinds of yoghurt were used : traditional yoghurt (TY), made with Streptococcus thermophilus and Lactobacillus bulgaricus starter cultures, and 'bifido' yoghurt (BY), made with the two starter cultures plus Bifidobacterium bifidum . After 7 d E. coli O157 : H7 decreased from 3·52 to 2·72 log₁₀ cfu ml⁻¹ and from 7·08 to 5·32 log₁₀ cfu ml⁻¹ in TY, and from 3·49 to 2·73 log₁₀ cfu ml⁻¹ and from 7·38 to 5·41 log₁₀ cfu ml⁻¹ in BY. The pH values of yoghurt dropped from 6·6 to 4·5 and 4·4 in TY (for low and high pathogen inocula, respectively), and from 6·6 to 4·6 and 4·5 in BY (for low and high pathogen inocula, respectively).     

The rapid estimation of microbial contamination of raw meat by measurement of adenosine triphosphate (ATP)

Bacteria were separated from raw meat homogenate by a simple three-stage process. Centrifugation (10 s at 2000 g) removed coarse particles; stirring with the cation exchange resin Bio-Rex 70 removed smaller particles and filtration through 0.22 μm membranes removed soluble materials. By this process 70—80% of the microbial populations of meat homogenates were consistently isolated on the filters. A linear relationship was found between log₁₀ microbial ATP and log₁₀ colony count of meat over the range 10⁵—10⁹ cfu/g. The value of ATP/cfu for meat samples was within the range previously reported for pure cultures. These data indicated that ATP extracted from the filters originated from bacteria in the meat samples. Several samples can be analysed simultaneously in an elapsed time of 20—25 min. The variability associated with estimates of both colony counts and ATP levels has been determined.     

Stability of genetic structure and effective population size inferred from temporal changes of microsatellite DNA polymorphisms in the land snail Helix aspersa (Gastropoda: Helicidae)

Temporal evolution of genetic variability may have far-reaching consequences for a diverse array of evolutionary processes. Within the polders of the Bay of Mont-Saint-Michel (France), populations of the land snail Helix aspersa are characterized by a metapopulation structure with occasional extinction processes resulting from farming practices. A temporal survey of genetic structure in H . aspersa was carried out using variability at four microsatellite loci, in ten populations sampled two years apart. Levels of within-population genetic variation, as measured by allelic richness, H _e or F _is , did not change over time and similar levels of population differentiation were demonstrated for both sampling years. The extent of genetic differentiation between temporal samples of the same population established (i) a stable structure for six populations, and (ii) substantial genetic changes for four populations. Using classical F -statistics and a maximum likelihood method, estimates of the effective population size ( N _e) illustrated a mixture of stable populations with high N _e, and unstable populations characterized by very small N _e estimates (of 5–11 individuals). Owing to human disturbances, intermittent gene flow and genetic drift are likely to be the predominant evolutionary processes shaping the observed genetic structure. However, the practice of multiple matings and sperm storage is likely to provide a reservoir of variability, minimizing the eroding genetic effects of population size reduction and increasing the effective population size.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 89–102.     

Bacteriological evaluation of groups of beef carcasses before the wash at six Alberta abattoirs

K.W.F. JERICHO, J.A. BRADLEY AND G.C. KOZUB. 1994. A method has been developed for the bacteriological evaluation of groups of beef carcasses which can be used to measure the degree of control over hygiene during hide removal and carcass dressing in abattoirs. This method, which enumerates aerobic mesophilic bacteria automatically using a hydrophobic grid membrane filter, was applied at six abattoirs. Two hundred excision samples (5 × 5 × 0.5 cm) were taken at 10 sites on the external surface of a group of 20 carcasses (five carcasses were sampled on each of four consecutive daily visits) for group-carcass evaluation at each abattoir. For each abattoir, the mean log₁₀ Most Probable Number of Growth Units (MPNGU) and between-carcass variance component were obtained for each site and the average over sites. Using the average within-abattoir variance of this study and previously published studies involving 76 additional carcasses (Jericho et al. 1993), it was determined that 20 carcasses are more than adequate to estimate the mean log₁₀ MPNGU per cm² within 0.5 units at a site. The distribution of the log₁₀ MPNGU per cm² over the 10 sites was compared for the abattoirs, and sites were found to cluster into 2–4 homogenous groups. The means over sites of log₁₀ MPNGU per cm² for the abattoirs ranged from 1.52 to 2.64 and were unrelated to line speed     

Reduction of Brochothrix thermosphacta on beef surfaces following immobilization of nisin in calcium alginate gels

Lean and adipose beef carcass tissues inoculated with Brochothrix thermosphacta (BT) (approx. 4.50 log₁₀ cfu cm⁻²) were left untreated (U) or treated with 100 μg ml⁻¹ nisin (N), calcium alginate (A) or 100 μg ml⁻¹ nisin immobilized in a calcium alginate gel (AN). Tissue samples were refrigerated after treatments and bacterial populations and nisin activity were determined at 0, 1, 2 and 7 d. U, A and N treatments of lean and adipose tissues did not suppress bacterial growth (>6 log₁₀ cfu cm⁻² by day 7) while treatments of lean and adipose tissues with AN suppressed bacteria (>2.42 log₁₀ cfu cm⁻² by day 7). Bacteriocin titres from both tissues were higher in AN vs N samples after the 7 d incubation. This study demonstrates that immobilization of nisin in a gel may be a more effective delivery system of a bacteriocin to the carcass surface than direct application.     

A note on estimation of food spoilage yeasts by measurement of adenosine triphosphate (ATP) after growth at various temperatures

The growth at 4, 10 and 15C of six psychrotrophic yeasts isolated from foods was compared using total viable counts and ATP measurement. A linear relationship ( r > 0.97) was obtained between log₁₀ (number of viable yeasts) and log₁₀ (ATP content) of cultures grown at these temperatures. This relationship was not temperature-dependent. The results are discussed and their significance for the rapid estimation of yeasts in foods is considered.     

The environmental determinants of the insular butterfly faunas of the British Isles

A multiple regression analysis was performed upon selected environmental variables for a series of islands in the British Isles, to establish their effects upon the size of the butterfly fauna, measured as he number of species regularly breeding, S_B .
So that the data be normally distributed, the regression analyses were performed upon log₁₀ transformed data only, with the data for outliers, mainland Britain and Ireland, the two largest islands, excluded.
Most highly correlated with the number of butterfly species breeding upon an island is the number breeding within a 25 km radius of the nearest point of the mainland, r ²=0.5941, followed by the correlations with the latitude of the mid-point of the island, r ²=0.5541, the number of plant species comprising the island Hora, r ²=0.5225, and the distance separating the island from the mainland, r ²=0.4514.
A partial correlation analysis confirms the importance of the parameters distance separating the island from the mainland, D ₁, and the size of the faunal source S _F , and rejects the importance of the size of the flora and the latitude of the island. This is further confirmed by the results of a step-wise regression analysis, the two variables D ₁ and S_F accounting for 66% of the variation of the butterfly fauna.
If an alternative measure of isolation, D ₂, which allows for the geographical clumping of islands, is combined with the variable S_F , then 69% of the variation of the butterfly fauna is accounted for.     

Use of impedance measurements to estimate numbers of antibiotic resistant Salmonella strains

Impedance detection times were compared with traditional plating methods for enumerating antibiotic-resistant strains of Salmonella stanley, Salm. thompson and Salm. infantis grown in laboratory medium and pork slurry. The correleation of log₁₀ counts of salmonellas with detection times was highly significant ( r = -0·96 for broth and r = -0·94 for slurries. The confidence limits (± og₁₀ 1·0 for broth and ± log₁₀ 1·65 for slurry) indicated that detection times could reliably be used as a rapid means of enumerating salmonellas when large numbers of counts of known strains are required for growth studies. Use of antibiotic-resistant strains also permitted their selective detection by impedance from the natural spoilage flora of pork slurry when the same antibiotics were incorporated in the detection medium.     

Comparison of conductimetric and traditional plating techniques for detecting yeasts in fruit juices

Frozen fruit juice concentrates containing an average microbial population of log₁₀ 1.54 cfu ml^-1 were examined by traditional plating techniques and direct and indirect conductimetry. The initial populations in diluted (1:4) concentrates increased to an average of log₁₀ 3.82 cfu ml^-1 during incubation at 25°C for 24 h. Incubation before plating and subjecting to conductimetric tests also facilitated the resuscitation of cells that may have been freeze-injured. Yeasts were recovered in equal numbers on acidified (pH 3.5) potato dextrose agar and dichloran rose bengal chloramphenicol agar (pH 5.6). Yeasts and bacteria were recovered on orange serum agar. Detection times determined by indirect conductimetry correlated fairly well ( r = -0.73) with populations (cfu ml^-1) detected on traditional plating media. Populations in diluted concentrates which were not incubated before examination were detected conductimetrically in an average of 48.9 h, whereas detection times for diluted concentrates incubated for 24 h at 25°C before testing were reduced to an average of 14.1 h. Examination by conventional (direct) conductimetry required an additional 10–20 h to reach changes in conductance of 5 μS h^-1.     

Rapid estimation of Escherichia coli in live marine bivalve shellfish using automated conductance measurement

Conductance measurement for quantitative estimations of Escherichia coli in live bivalve shellfish was evaluated as an alternative to the conventional most probable number (MPN) method used in France. The sensitivity of the two techniques was comparable. A single regression line ( r =-0·968, P < 10^-6) between log₁₀ MPN and detection time (DT) was used to estimate E. coli concentrations for all shellfish examined. Estimation accuracy was ± 0·92 log unit. Repeatability was better for DT than the log₁₀ of MPN estimations (average coefficients of variation 2·7 and 9·3%, respectively). The conductance signal was attributable to E. coli in 96% of cases, and only 0·7% of E. coli cultures failed to exhibit a signal within 20 h. The conductance method reduces analysis handling time and is much easier to use than the MPN method. Moreover, results can be obtained within 5–9 h compared to 3 d for the MPN method.     

Measurement of virulence of aeromonads using a suckling mouse model of infection

Abstract Virulent and avirulent strains of Aeromonas spp. were identified and virulence quantified using an animal model. Virulence was measured by determining a 50% lethal dose (LD₅₀) 43 h after oral administration of live bacteria. The LD₅₀ of virulent Aeromonas isolates ranged from log₁₀ 7.53 (mean) organisms to log₁₀ 8.88 (mean). Some isolates were avirulent in this model. Detection of cytotoxic activity in culture supernatants correlated with virulence (Fisher exact test, P = 0.0029). There was no correlation between LD₅₀ and the source of the isolate, β-haemolysis or lipopolysaccharide (LPS) banding profile on SDS-PAGE. In this animal model, virulence was multifactorial in that: (i) bacterial multiplication in the gut was associated with fatal infection; (ii) the increase in bacterial numbers in the gut of mice administered a lethal dose of bacteria was accompanied by accumulation of fluid; and (iii) there was evidence of extraintestinal spread of infection. Protection of suckling mice by rabbit antiserum to Aeromonas cell envelopes was observed.     

Comparison of gas use efficiency and treatment uniformity in a forest ecosystem exposed to elevated [CO2] using pure and prediluted free-air CO2 enrichment technology

A direct comparison of treatment uniformity and CO₂ use of pure and prediluted free-air CO₂ enrichment (FACE) systems was conducted in a forest ecosystem. A vertical release pure CO₂ fumigation system was superimposed on an existing prediluted CO₂ fumigation system and operated on alternate days. The FACE system using prediluted CO₂ fumigation technology exhibited less temporal and spatial variability than the pure CO₂ fumigation system. The pure CO₂ fumigation system tended to over-fumigate the upwind portions of the plot and used 25% more CO₂ than the prediluted CO₂ fumigation system. The increased CO₂ use by the pure CO₂ system was exacerbated at low wind speeds. It is not clear if this phenomenon will also be observed in plots with smaller diameters and low-stature vegetation.