Salmonella in animal slurry can be destroyed by aeration at low temperatures

Cattle and other animals infected by Salmonella can emit high numbers of these bacteria. To determine an effective means for reducing this bacterial group in animal slurry, samples were subjected to aeration in laboratory experiments and in farm-scale slurry tanks. A clear reduction in Salmonella levels was found in laboratory experiments at temperatures from 4 to 40 °C. Aeration in farm-scale slurry tanks increased the temperature above the ambient temperatures (often less than 0 °C) to maxima ranging between 19 and 40 °C. Farm-scale aeration resulted in similar reductions in Salmonella as those achieved in laboratory experiments. Thus, reductions, ranging from greater than 99% of the initial number to no detectable Salmonella , could be reached after 2–5 weeks using aeration processes with cattle slurries contaminated by Salm. infantis or pig slurry contaminated by Salm. typhimurium . These results suggest that farmers can control the spread of Salmonella from slurry to agricultural fields. The reduction mechanisms remain unknown, though the increase in pH (to 7·6–9·0) found in slurries after aeration might exert a decreasing effect on these bacteria.     

Heat-resistant Bacteria in Pasteurized Whole Egg

Samples of egg melange taken from an egg packing station contained an average of 7·3 x 10⁴ organisms/ml which survived laboratory pasteurization at 65°C for 3 min. Many of the organisms surviving pasteurization were found to be coryneform bacteria related to Microbacterium lacticum which could be differentiated into several groups. The remainder were a miscellaneous collection of unidentified cocci and coccobacilli and some Bacillus spp. The coryneform bacteria were shown to be the most heat-resistant isolates with negligible loss of viability after 60 min at 65°C, At least two of the representative strains were very heat-resistant, 0·01% surviving 20 and 38 min at 80°C in phosphate buffer at pH 7·1. Growth tests showed that none of the isolates grew at 5°C after 10 d incubation but those capable of growing most rapidly at 10° and 15°C were also the most heat-resistant. Such strains had a doubling time at 15°C of between 6 and 8 h in whole egg. Freezing the coryneform bacteria in liquid whole egg at –18°C had negligible effect on viability or heat-resistance at 65°C.     

Bactericidal effect of hydrogen peroxide on Salmonella typhimurium in liquid whole egg

The effect of hydrogen peroxide on Salmonella typhimurium in whole egg was evaluated. The bactericidal effects observed on the test organism at 5° and 20°C were found to be similar. There was a 99% kill in the presence of 0°5% and 1°0% H₂O₂. Addition of the test organism and H₂O₂ after pre-heating the egg material at 40°C for 15 min caused a rapid kill which was 10000-fold greater than that produced by H₂O₂ alone.     

Inactivation of Salmonella enterica serovar enteritidis in shell eggs by sequential application of heat and ozone

Aims:  To assess the contribution of ozone to lethality of Salmonella enterica serovar Enteritidis in experimentally inoculated whole shell eggs that are sequentially treated with heat and gaseous ozone in pilot-scale equipment.
Methods and Results:  Whole shell eggs were inoculated with small populations of Salmonella Enteritidis (8·5 × 10⁴–2·4 × 10⁵ CFU per egg) near the egg vitelline membrane. Eggs were subjected to immersion heating (57°C for 21 min), ozone treatment (vacuum at 67·5 kPa, followed by ozonation at a maximum concentration of approx. 140 g ozone m⁻³ and 184–198 kPa for 40 min) or a combination of both treatments. Survivors were detected after an enrichment process or enumerated using modified most probable number technique. Ozone, heat and combination treatments inactivated 0·11, 3·1 and 4·2 log Salmonella Enteritidis per egg, respectively.
Conclusions:  Sequential application of heat and gaseous ozone was significantly more effective than either heat or ozone alone. The demonstrated synergy between these treatment steps should produce safer shell eggs than the heat treatment alone.
Significance and Impact of the Study:  Shell eggs are the most common vehicle for human infection by Salmonella Enteritidis. Many cases of egg-related salmonellosis are reported annually despite efforts to reduce contamination, including thermal pasteurization of shell eggs and egg products. Treatment with ozone-based combination should produce shell eggs safer than those treated with heat alone.     

Effects of essential oil from mint (Mentha piperita) on Salmonella enteritidis and Listeria monocytogenes in model food systems at 4° and 10°C

The effect of mint ( Mentha piperita ) essential oil (0·5, 1·0, 1·5 and 2·0%, v/w) on Salmonella enteritidis and Listeria monocytogenes in a culture medium and three model foods; tzatziki (pH 4·5), taramosalata (pH 5·0) and pâté (pH 6·8), inoculated at 10⁷ cfu g^-1, at 4° and 10°C for ca 1 week was studied. In the culture medium supplemented with the essential oil, no growth was observed over 2 d at 30°C determined by a conductance method with a Malthus 2000 growth analyser. Salmonella enteritidis died in tzatziki in all treatments and declined in the other foods except for pâté at 10°C as judged with viable counts. Listeria monocytogenes populations showed a declining trend towards the end of the storage period but was increased in pâté. Mint essential oil antibacterial action depended mainly on its concentration, food pH, composition, storage temperature and the nature of the micro-organism.     

Immersion heat treatments for inactivation of Salmonella enteritidis with intact eggs

The effects of water-bath immersion heat treatments on the inactivation of Salmonellaenteritidis within intact shell eggs were evaluated. Six pooled strains of Salm. enteritidis ( ca 3×10⁸ cfu, inoculated near the centre of the yolk) were completelyinactivated within 50–57·5 min at a bath temperature of 58°C and within 65–75min at 57°C (an 8·4 to 8·5- D process per egg). Following the initial 24 to35-min come-up period, semilogarithmic survivor curves obtained at 58 and 57°C yieldedapparent decimal reduction times ( D -values) of 4·5 and 6·0 min, respectively.Haugh unit values increased during heating, while yolk index and albumen pH values wereunaffected. Albumen clarity and functionality were affected by the thermal treatments; therefore,extended whip times would be required for meringue preparation using immersion-heated eggwhites. Immersion-heated shell eggs could provide Salmonella -free ingredients for thepreparation of a variety of minimally-cooked foods of interest to consumers and foodserviceoperators.     

Effect of temperature and water activity on in vitro germination of Monilinia spp.

Aims:  This study evaluated the effect of temperature (0–38°C) and water activity ( a _w: 0·87–0·99) on the lag phase prior to germination and the percentage of germination over time for Monilinia laxa , Monilinia fructicola and Monilinia fructigena .
Methods and Results:  More than 80% of viable conidia germinated at 25°C and 0·99 a _w within 2 h for M. fructicola and M. fructigena and 4 h for M. laxa . There was no germination at 38°C, and all three Monilinia spp. germinated at 0°C. At the lowest a _w (0·87), none of the Monilinia spp. was able to germinate at any of the incubation temperatures studied. Whereas at 0·90 a _w, conidia were only able to germinate at 15, 25 and 30°C for the three species studied, except for M. fructicola at 15°C. In contrast, at 0·95, 0·97 and 0·99 a _w, germination occurred at all studied temperatures less 38°C. Generally, the lag phase was longer at low levels of a _w (0·90–095), and differences were more evident as temperatures were far from the optimum (0–5°C).
Conclusions:  Germination and lag phase period were markedly influenced by temperature and a _w, and in general when conditions of temperature and a _w were suboptimal, the lag phase was longer and the percentage of germination was lower.
Significance and Impact of the Study:  Knowledge of the germination requirements of this fungus is important in order to understand their behaviour in natural situations and to provide baseline data required for the construction of new prediction models. Our study might be used to develop a predictive model to understand and control the disease caused by Monilinia spp.     

Influence of acidity and initial substrate temperature on germination of Rhizopus oligosporus sporangiospores during tempe manufacture

J.C. DE REU, F.M. ROMBOUTS AND M.J.R. NOUT. 1995. During the soaking of soya beans according to an accelerated acidification method organic acids were formed, resulting in a pH decrease from 6·0 to 3·9. After 24 h of fermentation at 30°C, lactic acid was the major organic acid (2·1% w/v soak water), while acetic acid (0·3% w/v soak water) and citric acid (0·5% w/v soak water) were also found. During cooking with fresh water (ratio raw beans: water, 1: 6·5) the concentrations of lactate/lactic acid and acetate/acetic acid in the beans were reduced by 45% and 51%, respectively.
The effect of organic acids on the germination of Rhizopus olgosporus sporangiospores was studied in liquid media and on soya beans. Germination in aqueous suspensions was delayed by acetic acid: within 6 h no germination occurred at concentrations higher than 0·05% (w/v incubation medium), at pH 4·0. When soya beans were soaked in the presence of acetic acid, the inhibitory concentration depended on the pH after soaking. Lactic acid and citric acid enhanced germination in liquid medium, but not in tempe.
Inoculation of soya beans with R. oligosporus at various temperatures followed by incubation at 30°C resulted in both increased and decreased periods for the lag phase of fungal growth. A maximum difference of 3 h lag phase was found between initial bean temperatures of 25 and 37°C.
When pure cultures of homofermentative lactic acid bacteria were used in the initial soaking process, less lactic acid and acetic acid was formed during soaking than when the accelerated acidification method was used. This resulted in a reduction of the lag phase before growth of R. oligosporus by up to 4·7 h.     

The survival of Salmonella enterica serovar Typhimurium DT104 and total viable counts on beef surfaces at different relative humidities and temperatures

Aims:  The aim of this study was to investigate changes in Salmonella and total viable count (TVC) survival on beef carcass surfaces stored for 72 h under different combinations of relative humidity (i.e. RH 75% or 96%) and temperature (5°C or 10°C).
Methods and Results:  The influence of low water activity ( a _w) and temperature on the survival and growth of Salmonella enterica serovar Typhimurium DT104 and the aerobic mesophilic flora on meat pieces from different sites on beef carcasses was investigated, under controlled conditions (75% or 96% RH; 5 or 10°C) in an environmental cabinet. Salmonella counts declined during storage at low a _w (75% RH) conditions at 5°C or 10°C. Salmonella counts increased during storage at high a _w (96% RH) at 10°C only. At 5°C, TVCs increased during storage at high a _w, but not at low a _w. TVCs increased on all samples from carcasses stored at high or low a _w at 10°C, except those samples taken from areas of surface fat.
Conclusions:  This suggests that substrate composition dictates growth rates under low a _w conditions. The results are discussed in terms of the possible protective effects of substrate osmolyte accumulation in bacterial survival and/or growth.
Significance and Impact of the Study:  The data obtained in this study provides useful insights on the influence of a _w and temperature on pathogen survival on meat surfaces at chill temperature.     

METHODS OF ASSESSING THE SPORICIDAL EFFICIENCY OF AN ULTRA-HIGH-TEMPERATURE MILK STERILIZING PLANT. II. EXPERIMENTS WITH SUSPENSIONS OF SPORES IN MILK

SUMMARY: The sporicidal efficiency of an ultra-high-temperature (UHT) milk processing plant has been tested using spores of a strain of Bacillus subtilis in milk. With the inoculum and volume of milk adjusted to obtain a countable number of survivors by a conventional dilution counting method, a temperature of 130·5° with the minimum time setting of the plant was found necessary to give a destruction of 99·99999%. This temperature was lower than that found previously (135°) for spores suspended in water and evidence is produced to support the suggestion that UHT milk may be inhibitory to the germination and/or subsequent growth of heated spores.     

Effect of Maillard reaction products on the growth of selected food-poisoning micro-organisms

The activity of the Maillard reaction products (MRP) prepared by heating (15 h at 90°C) a solution of 1·71 mol/l glucose and 2·05 mol/l glycine at pH values 6·0 and 8·8, against food-poisoning micro-organisms, including Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium, Salmonella enteritidis and Aeromonas hydrophila , was investigated. High and low pH MRPs strongly inhibited A. hydrophila , whereas Staph. aureus and L. monocytogenes were slightly inhibited by the high pH MRPs only and Salmonella strains were resistant to both.     

Factors affecting the survival of Salmonella and Escherichia coli in anaerobically fermented pig waste

The survival of Salmonella typhimurium was investigated in acidogenic, anaerobically fermented pig wastes and in synthetic media, each containing volatile fatty acids (VFA). Salm. typhimurium survived at pH 6·8, but not at pH 4·0, when incubated at 37°C for 24 h in either fermented or synthetic medium containing VFA. The minimum inhibiting concentration of VFA for Salm. typhimurium after 48 h incubation at 30°C at pH 4·0 was 0·03 mol/l and for Escherichia coli it was 0·09 mol/l. Fermented pig wastes in a digester, maintained at pH 5·9, were inoculated with Salm. typhimurium and then incubated at 37°C for 24 h. The pH was adjusted to either 4·0 or 5·0 and after a further 48 h at 30°C, Salm. typhimurium survived at pH 5·0 but not at pH 4·0. It was concluded that pH is critical in determining the survival of this organism in acidogenic anaerobically fermented pig waste.     

THE PREVENTION OF MICROBIAL SPOILAGE IN WHOLE SHELL EGGS BY HEAT TREATMENT METHODS

SUMMARY: The prevention of microbial spoilage in whole shell eggs by heat treatment methods has been investigated. After shell eggs had been immersed for 3 min in water at 144·5 ± 0·5°F the incidence of microbial rotting was reduced by over 90%. Consistent reductions in spoilage losses were obtained with farm steeped eggs, eggs produced under commercial conditions, and eggs held in cold storage for 6 months. Heat treatment efficiently controlled green, sour and black rots in stored eggs, but although reductions were noted in the number of mould rots, it cannot be guaranteed that this type of spoilage can be controlled with similar certainty.
Internal quality in heat treated eggs was satisfactory. During protracted holding in cold storage, heat treatment appeared to result in a better internal egg quality and a more stable keeping quality, than when untreated eggs were held under similar conditions.     

Efficiency of different enrichment and isolation procedures for the detection of Salmonella serotypes in edible offal

Rapid detection systems for Salmonella in foodstuffs are currently being developed. However, existing standards still call for application of traditional methods employing pre-enrichment followed by selective enrichment and isolation. The efficacy of various methods was tested using 264 chicken and lamb organ meats. Pre-enrichment was carried out in Tryptone Soy Broth (TSB) and enrichment in Tetrathionate Brilliant Green Broth (TTB) at 37°C, Selenite Broth with Brilliant Green and Sulphapyridine at 37°C and 43°C, and Rappaport-Vassiliadis Broth (RV 10) at 42°C. The isolation media were Brilliant Green Agar (BGA), Deoxycholate Citrate Agar, Hektoen Enteric Agar (HEA) and Salmonella-Shigella Agar.
Enrichment in RV/42°C followed by isolation on BGA as recommended by ISO standard no. 6579 and enrichment in TTB/37°C followed by isolation in HEA, no longer recommended by that standard, produced the best results. Low percentages of positive samples and difficulties in detecting Salmonella are the result of interference by competing organisms (Enterobacteriaceae) and the number of salmonellas present after enrichment.
A total of 528 samples (TSB, eggs, lamb liver and chicken liver) were inoculated with Salm. enteritidis, Salm. kapemba and Salm. virchow , and the preceding experiment was repeated. All the TSB and egg samples tested positive, but the percentage of positive samples from the lamb and chicken liver was only 81–92%. Recovery of the salmonellas did not depend upon the method employed or the serotype inoculated but instead on interference by competing flora and the numbers of Salmonella present in the samples.     

Autotriploid tench Tinca tinca (L.) larvae obtained by fertilization of eggs previously subjected to postovulatory ageing in vitro and in vivo

Eggs of diploid tench Tinca tinca were half-stripped out and stored for 0 (control batch), 1, 3 and 5 h at mean ± s . d . 17·0 ± 0·4 and 21·9 ± 0·5° C or for 0, 1, 2, 3, 4 and 5 h at 24·0 ± 0·0° C in vitro prior to fertilization. The eggs remaining in vivo in the fish kept at 17·0 ± 0·4 and 21·9 ± 0·5° C were collected and fertilized in the same time intervals. Fertilization rate and larval yield mostly decreased after 3–5 h storage of eggs both in vitro and in vivo and only the diploid larvae were found in all control batches. Triploid larval yields increased to a maximum 5·26% after 5 h in vitro storage at 24·0° C and 1·07 and 1·60% after 3 h in vitro storage at 21·9 and 17·0° C, respectively. Triploid larval yield during in vivo storage at 21·9° C reached a maximum 0·91% after 5 h. As the spontaneous autotriploid larvae arose solely from fertilized eggs previously subjected to postovulatory egg ageing by means of prolongated storage, the autotriploidy was probably caused by failure of extrusion of the second polar body.     

The effect of prior heat shock on the thermoresistance of Salmonella thompson in foods

The resistance of Salmonella thompson to heating at 54° or 60°C in tryptone soya broth, liquid whole egg, 10% or 40% reconstituted dried milk or minced beef was increased if cells were held at 48°C for 30 min before heating at the higher temperatures. Induction of thermotolerance by mild heat shock is thus not confined to cells grown and heated in broth systems. The heat shock phenomenon may therefore have implications for the safety of foods given marginal heat treatment.     

The influence of temperature on the life cycle and occurrence of three species of lake-dwelling leeches (Annelida: Hirudinea)

The influence of constant temperatures on the growth, maturation and reproduction of field-collected and F, generation Helobdella stagnalis, Glossiphonia complanata and Erpobdella octoculata was investigated. Leech material from a lowland, productive lake and from an upland, unproductive lake was cultured at each of the temperatures: l·5,5·5,8, 10·5, 13,16, 19 and 23°C. Field-collected and F₁ generation H. stagnalis and E. octoculata matured and produced eggs and young over the range 10·5–23°C, whereas the range for egg production was 8–23°C for field-collected G. complanata, F, generation leeches failing to mature at any temperature due to an additional limiting factor being involved.
Optimal temperatures for cocoon/egg production were observed for E. octoculata, but egg production was similar at all temperatures for G. complanata and H. stagnalis. In contrast, the viability of eggs was greater at certain temperatures for the last two species but was similar at all temperatures for E. octoculata. Differences in egg production between field-collected and F, generation leeches are discussed. Rising temperatures accelerated rates of egg hatching, and of growth and maturation of leeches. The observed differences in survival of leeches at the various temperatures are discussed. Ecotypic adaptation to temperature within the small geographical area of Britain may not occur. It is concluded that though temperature may be a contributory factor determining distribution and abundance in British lakes, other causative factors, such as the occurrence and abundance of food organisms, are more likely to be implicated.     

Cry (preservation of spermatozoa of the whitefish Coregonus muksun Pallas

The cryopreservation methods for whitefish, Coregonus muksun , spermatozoa were studied using six different extenders with -40°C and -196°C as storing temperatures. In 1980, the spermatozoa of individual fishes were frozen in dry ice, in vials containing three different media based on glycerol as cryoprotectant. The pure glycerol media resulted in a 20% average rate of fertilization when samples were stored at — 40°C. The thawing procedure did not affect fertilization. In 1981, the sperm of individual fishes was frozen in pellets and stored in liquid nitrogen. Cryopreserved semen thawed in 0·119M NaHCO₃ produced 29·6–87·7% eyed eggs. The differences were caused mainly by the type of extender. The highest rate of fertility was obtained using an extender consisting of 0·3 M glucose with 20% glycerol (87·7% eyed eggs; 98·6% eyed eggs when compared to the corresponding control). The same values, using the modified extender of Stoss (Stoss & Refstie, in press), were 38·7–82·4% and, on the average, 90% eyed eggs, respectively.     

Modelling the growth of Weissella cibaria as a function of fermentation conditions

Aims:  To investigate the effect of pH, water activity ( a _w) and temperature on the growth of Weissella cibaria DBPZ1006, a lactic acid bacterium isolated from sourdoughs.
Methods and Results:  The kinetics of growth of W. cibaria DBPZ1006 was investigated during batch fermentations as a function of pH (4·0–8·0), a _w (0·935–0·994) and temperature (10–45°C) in a rich medium. The growth curve parameters (lag time, growth rate and asymptote) were estimated using the dynamic model of Baranyi and Roberts (1994. A dynamic approach to predicting bacterial growth in food. Int J Food Microbiol 23, 277–294). The effect of pH, a _w and temperature on maximum specific growth rate (μ_max) were estimated by fitting a cardinal model. μ_max under optimal conditions (pH = 6·6, a _w = 0·994, T  = 36·3°C) was estimated to be 0·93 h⁻¹. Minimum and maximum estimated pH and temperature for growth were 3·6 and 8·15, and 9·0°C and 47·8°C, respectively, while minimum a _w was 0·918 (equivalent to 12·2% w/v NaCl).
Conclusions:  Weissella cibaria DBPZ1006 is a fast-growing heterofermentative strain, which could be used in a mixed starter culture for making bread.
Significance and Impact of the Study:  This is the first study reporting the modelling of the growth of W. cibaria , a species that is increasingly being used as a starter in sourdough and vegetable fermentations.     

Temperature-dependent development rates of cod Gadus morhua eggs

Temperature development relationships were determined for batches of Irish Sea cod Gadus morhua eggs incubated in flow-through incubators. Hatching began 16·4 days after fertilization (DAF) at 6° C, 10·3 DAF at 8° C, 9·4 DAF at 10° C and 7·4 DAF at 12° C. Egg mortality increased at the higher temperatures, but survival was >80%. Results were compared with published data at four comparable stage end points: the end of blastula, the end of gastrula, the point of growth of the embryo completely surrounding the yolk and the point when 50% of the eggs were hatched. All the studies showed a curvilinear relationship between age at stage and temperature. There was a 12 day inter-study difference in time to 50% hatch at 2° C and 4 day difference at 10° C. There were no consistent trends that differentiated eastern v. western, or northern v. southern populations. A single model for cod egg incubation time from fertilization to 50% hatch was derived based on data from six cod populations, but it is recommended that individual stock relationships should be used where possible.