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.     

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.     

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     

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.     

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.     

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.     

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.     

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.     

The variability of orchid population size

Orchid population size has long been believed to be highly variable in time and space. Here we present the first quantitative assessment of the variability of orchid population sizes and compare them with plant and animal population data from the literature. Three measures of temporal variation were used (standard deviation of log₁₀ (population size), Hurst exponent and variance in the intrinsic rate of increase). Eight unfertilized plots of Orchis morio in a factorial fertilizer experiment were assessed for variability in population size over a 26 year period. The Hurst exponent across the eight plots gave measures consistent with pink noise well into black noise (0.86 to 1.87). The variation in the intrinsic rate of increase suggested that the expected time to extinction in the plots differed by an order of magnitude. Standard deviation of log₁₀ (population size) across plots was greater than the corresponding temporal variation within plots. Standard deviation of log₁₀ (population size) reached an asymptote between 10 and 20 years. The range of temporal variation between plots in this and other orchid populations was as great as the mean values reported for birds, insects and mammals. These results have highlighted the importance of spatial variation: until we know the relative importance of spatial and temporal variation for more plant species we will be unable to assess whether orchid populations are more variable in size than other species.     

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.     

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.     

Cannibalism on eggs by dark chub, Zacco temmincki (Temminck & Schlegel) (Cyprinidae)

During spawning male dark chub, Zacco temmincki (Temminck & Schlegel), buried eggs by stirring up the river bed with their anal fins. No parental care was shown for eggs after spawning. Many satellites (both males and females) preyed upon eggs at the instant of spawning. The number of eggs produced is a function of body length [log₁₀ E =−0.273+3.519log₁₀ S , where E is the number of eggs and S is standard length (cm)]. From the size distribution of females that spawned during a reproductive season, the total number of spawning sites used and mean number of eggs per spawning site, at least 97.1% eggs were estimated to be consumed by cannibalism. This marked cannibalism resulted from a lack of parental care and the presence of many satellites produced by the polygynous mating system.     

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.     

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 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.     

Estimation of the growth rates of epiphytic bacteria and Lemna minor in a river

SUMMARY. Growth of Lemna minor fronds in the River Frome during summer was found to be logarithmic with time and the growth rate (log₁₀) was 0.066 day⁻¹. This is equivalent to a doubling time of 4.5 days. The life expectancy of the fronds was 34 days.
The net change in the density of bacteria epiphytic on the lower surface of Lemna fronds in the R. Frome was monitored using a direct microscopic technique. The observed increase in bacterial numbers has been partitioned into the components of attachment and growth, assuming that attachment occurred at a constant rate and that the bacterial population grew logarithmically. The line which fitted the data best gave an attachment rate of 5.7 × 10⁵ bacteria cm⁻² day⁻¹ and a growth rate (log₁₀) for the bacteria of 0.044 day⁻¹ which is equivalent to a doubling time of 164 h.
Estimates of the rate of detachment of bacteria from Lemna plants were obtained from a laboratory experiment which assumed negligible growth of bacteria in 1 h. The number of bacteria which detached per hour and the sizes of the bacterial populations on the plants before and after detachment were estimated using a plating technique. Different detachment rates were monitored. The detachment rate (analogous to growth rate) which is judged to be most similar to an in situ value was 0.0031 h⁻¹ (0.074 day⁻¹). This rate added to the specific growth rate given above resulted in a corrected growth rate of 0.118 day⁻¹ equivalent to a doubling time of 61 h.     

Clostridia spore formation during aerobic deterioration of maize and sorghum silages as influenced by Lactobacillus buchneri and Lactobacillus plantarum inoculants

Aims:  The effect of the inoculation of maize and sorghum silages with Lactobacillus plantarum (LP) and Lactobacillus buchneri (LB) on the clostridia spore formation during aerobic deterioration has been studied.
Methods and results:  The crops were ensiled in 30 l jars, without a lactic acid bacteria inoculant (C), and with an LP or LB inocula (theoretical rate of 1 × 10⁶). After 90 days of conservation, the silages were analysed for the chemical and microbiological characteristics and subjected to an aerobic stability test, during which pH, temperature, nitrate, yeast, mould and clostridia spores were measured. Compared to the C and LP silages, yeasts were reduced in the LB silages, resulting in an increased aerobic stability. Clostridia spores, determined by most probable number (MPN) procedure, increased to 6 log₁₀ MPN g⁻¹ in the C and LP maize silages, whereas they reached 3 log₁₀ MPN g⁻¹ in C and LP sorghum silages.
Conclusions:  Clostridia spore count only slightly increased in the LB maize silages after 342 h (2·59 log₁₀ MPN g⁻¹), whereas it did not show any increase in the LB sorghum silages for the whole period of air exposure.
Significance and impact of the study:  The data indicated that clostridia spore outgrowth can take place during silo feedout in aerobic-deteriorated silages and that LB inoculation reduces the risk of clostridia outgrowth after silage opening by increasing the aerobic stability.     

The influence of thermal parameters on the acclimation responses of pinfish Lagodon rhomboides exposed to static and decreasing low temperatures

Pinfish Lagodon rhomboides acclimation rates were determined by modelling changes in critical thermal minimum ( T _{crit min}, ° C) estimates at set intervals following a temperature decrease of 3–4° C. The results showed that pinfish gained a total of 3·7° C of cold tolerance over a range of acclimation temperatures ( T _acc, ° C) from (23–12° C), that cold tolerance increased with exposure time to the reduced temperature at all T _acc, but that the rate of cold tolerance accruement (mean 0·14° C day⁻¹) was independent of T _acc. A highly significant ( P < 0·001) multivariate predictive model was generated that described the acclimation rates and thermal tolerance of pinfish exposed to reduction in water temperature: log₁₀ T _{crit min}= 0·41597 − 0·01704 T _acc+ 0·04320 T _plunge− 0·08376[log₁₀ ( t + 1)], where T _plunge is plunge temperature (° C) and t is the time (days). A comparison of the present data, with acclimation rate data for other species, suggests that factors such as latitude or geographic range may play a more important role than ambient temperature in determining cold acclimation rates in fishes.     

THE INACTIVATION OF BACILLUS SUBTILIS SPORES IN PENICILLIN BY GAMMA RADIATION

SUMMARY: Sodium benzylpenicillin, contaminated with Bacillus subtilis spores by freeze-drying a suspension of spores in an aqueous penicillin solution ( c . 50% w/v), was exposed to gamma radiation and a 70-tube dilution method was used to determine the surviving spores after various doses. The correlation coefficient between log₁₀ percentage survival and dose was −0.9523. The regression of the former on the latter was calculated and the decimal reduction dose found to be 20.2 × 10⁴ rads. The regression and the decimal reduction dose were similar to those obtained when suspensions of spores in distilled water were irradiated.