Sensitivity of Listeria monocytogenes to irradiation on poultry meat and in phosphate-buffered saline

The sensitivity of four strains of Listeria monocytogenes to irradiation on poultry meat and in phosphate-buffered saline was investigated. The D₁₀ values on poultry meat were 0.417–0.553 kGy depending on strain and plating medium used. Lower values were obtained in phosphate-buffered saline. Generally tryptone soya yeast extract and McBride agars gave a better recovery (higher D₁₀ value) than listeria selective agar. The D₁₀ values for L. monocytogenes were similar to those reported for Salmonella spp. irradiated under similar conditions. Therefore irradiation doses suggested to eliminate salmonellas from poultry carcasses would also be sufficient to remove L. monocytogenes.     

Inactivation of Strains of Salmonella senftenberg by Gamma Irradiation

S ummary : Strains of Salmonella senftenberg isolated from Norwegian herring meal and strain 775W were exposed to gamma radiation from a ⁶⁰Co source. When they were irradiated in phosphate buffered saline solution, the average D₁₀ values for all the strains was 19·3 krad. On irradiating strains 56 and 775W in herring meal the D₁₀ values were 192·1 and 188·5 krad, respectively, thus indicating that the suspending medium had a great effect on the radiation resistance of the organisms. Radiation doses of the order of 0·8–1·3 Mrad are recommended for the decontamination of herring meal.     

Sensitivity of bacteria to irradiation on poultry meat under various atmospheres

The sensitivity of seven bacterial species inoculated on to sterile poultry meat and irradiated under various atmospheres (air, CO₂, vacuum and nitrogen) was investigated. The irradiation sensitivities of Streptococcus faecalis and Staphylococcus aureus were unaffected by atmosphere. The other micro-organisms ( Pseudomonas putida, Salmonella typhimurium, Escherichia coli, Moraxella phenylpyruvica and a Lactobacillus sp.) were more sensitive (their D₁₀ values decreased) when irradiated in atmospheres other than air. In general, a vacuum or CO₂ atmosphere during irradiation was found to have the most lethal effect. Plating media were found to have a significant effect on the recovery of irradiated E. coli and Salm. typhirium.     

Beta-galactosidase from Aureobasidium pullulans

A red-pigmented, radiation-resistant, Gram-negative, rod-shaped bacterium isolated from irradiated pork is described. The D₁₀ values in buffer solution and on pork mince are 3·45 and 5·05kGy respectively. The strain has been identified as a Deinobacter species.     

Radiation inactivation of some food-borne pathogens in fish as influenced by fat levels

The influence of low (0·39–1·1%), medium (4·25%) and high (7·1–32·5%) fat levels in fish on radiation inactivation of four food-borne pathogens was investigated. Cells of Listeria monocytogenes 036, Yersinia enterocolitica F5692, Bacillus cereus and Salmonella typhimurium at logarithmic phase were inoculated in 10% fish homogenates and subjected to gamma irradiation at ice temperature (0–1 °C) with doses ranging from 0·05 to 0·8 kGy. The radiation survival curves of L. monocytogenes and B. cereus were characterized by shoulders, while a tailing effect was depicted by cells of Y. enterocolitica and B. cereus . The D₁₀ values in kGy calculated on the exponential part of the curve ranged from 0·2 to 0·3, 0·15 to 0·25, 0·1 to 0·15 and 0·09 to 0·1 for L. monocytogenes 036, B. cereus, Salm. typhimurium and Y. enterocolitica F5692, respectively. This order (D₁₀) of radiation resistance of each organism was not affected by the fat content of the fish. Inoculated pack studies carried out separately with each pathogen in fatty (Indian sardine, 7·1%) and lean (Golden anchovy, 0·39%) fish showed no difference in their survival after exposure to 1 kGy and 3 kGy doses, which corroborated the above observation. The practical significance of these results in the application of the technology is discussed.     

Prediction, from temperature, of eyeing, hatching and 'swim-up' times for salmonid embryos

SUMMARY. 1. The literature contains a number of curves relating time (days) required for median hatch (=D₂) to water temperature (T,°C) for the eggs of several salmonid fishes. There are relatively few data on the relationships between time to median eyed (= D₁), days) or time to median swim-up (= D₃, days) and temperature.
2. From published data, over most of the range 0-9.5°C, approximate relationships are D,=0.5D₂ for Atlantic salmon (Salmo salar L.) and D3 = 1.7D₂ for eight species of Salmo and Oncorhynchus.
3. Field and hatchery tests suggest that these are useful empirical models for approximate prediction of D₁ and D₃ from D₂ for most salmonids.     

CELL PROLIFERATION IN THE CASTRATE MOUSE SEMINAL VESICLE IN RESPONSE TO TESTOSTERONE PROPIONATE

The cell proliferation kinetics following induced DNA synthesis in the mouse seminal vesicle were measured after treatment with testosterone propionate. Fraction labelled mitosis curves at 24, 48 and 72 hr after injection gave t ₂ values of 1·5, 2·0 and 1·8 hr respectively, and t _s values of 10·5, 8·0 and 8·0 hr. T _c measured 48 hr after stimulation was 17·5 hr. Growth fraction rose from 0·14 at 24 hr to 0·64 at 48 hr, and fell to 0·32 by 72 hr. A simple model is proposed in which the rise and fall of mitotic index and labelled index is determined by the 'cell distribution ratio'.     

OBSERVATIONS ON THE MICROFLORA OF MINCED CHICKEN MEAT IRRADIATED WITH 4 MeV CATHODE RAYS

SUMMARY: Experiments are described in which minced chicken meat, packed anaerobically, was irradiated at room temperature and in the frozen state with a wide range of doses of 4 MeV cathode rays. Sterility was achieved in 14 out of 15 samples which had received 2 × 10⁶ rads or more. Doses of 0·5 and 1·0 × 10⁶ rads allowed survival of a few bacteria/g, usually spore formers. Bacterial counts indicated an approximately logarithmic decrease in numbers at lower doses, while freezing reduced the bactericidal effect.
The storage life at 5° was prolonged only slightly by doses of 5 × 10⁴ and 10 × 10⁴ rads, and highly variable results were obtained with 17·5 × 10⁴ rads. A dose of 25 × 10⁴ rads, however, increased the storage life very considerably. The types of bacteria present initially, and after irradiation with low doses and storage at 5°, were studied. After storage for 12 days or more various types of nonsporing Gram-positive rods were predominant in almost all samples, both control and irradiated. Streptococci were also important where irradiation with 17·5 × 10⁴ and 25 × 10⁴ rads was followed by long storage.     

A rapid method for the determination of bacterial fatty acid composition

Heat treatment of spores of non-proteolytic strains of Clostridium botulinum at 75–90°C, and enumeration of survivors on a nutrient medium containing lysozyme gave biphasic survival curves. A majority of spores were inactivated rapidly by heating, and the apparent heat-resistance of these spores was similar to that observed by enumeration on medium without lysozyme. A minority of spores showed much greater heat-resistance, due to the fact that the spore coat was permeable to lysozyme, which diffused into the spore from the medium and replaced the heat-inactivated germination system. The proportion of heated spores permeable to lysozyme was between 0.2 and 1.4% for spores of strains 17B (type B) and Beluga (type E), but was about 20% for spores of strain Foster B96 (type E). After treatment of heated spores with alkaline thioglycolate, all were permeable to lysozyme. D-values for heated spores that were permeable to lysozyme (naturally and after treatment with thioglycolate) were: for strain 17B, D₈₅°C, 100 min; D₉₀°C, 18.7 min; D₉₅°C, 4.4 min; for strain Beluga, D₈₅°C, 46 min; D₉₀°C, 11.8 min; D₉₅°C, 2.8 min. The z-values for these spores of strains 17B and Beluga were 7.6°C and 8.3°C.     

STUDIES ON SOLUBLE PROTEINS RELEASED FROM THE SYNAPTIC VESICLES OF RAT BRAIN CORTEX

Abstract— The soluble proteins released from the synaptic vesicles of rat cerebral cortex were studied. One fraction (D₄) of these proteins was released in parallel with release of acetylcholine when synaptic vesicles were incubated at 37°C for 10 min in isotonic medium. Another fraction (Dj) was liberated from synaptic vesicles when their membranes were ruptured by mild treatment under hyposmotic conditions and freeze-thawing after release of D₁ fraction. Fractions D₁ and D₂ contained 12 and 9 per cent, respectively, of the total protein in the synaptic vesicles. Some properties of these fractions were investigated by zone electrophoresis and ultracentrifugation, and by measuring their binding capacities for [¹⁴C]acetylcholine and various enzyme activities related to acetylcholine metabolism.     

The effect of temperature fluctuations on oxygen consumption and ammonia excretion of underyearling Lake Inari Arctic charr

Underyearling Lake Inari Arctic charr Salvelinus alpinus were acclimated to 11·0) C for 3 weeks, and then one group was maintained at 11·0) C and others were exposed to 14·4) Cconst, 17·7) C_const or a diel fluctuating temperature of 14·3° C ± 1° C (14·3° C_fluc). Routine rates of oxygen consumption and ammonia excretion were measured over 10 days before the temperature change and over 31 days following the change. Measurements were made on fish that were feeding and growing. The temperature increase produced an immediate increase in oxygen consumption. There was then a decline over the next few days, suggesting that thermal acclimation was rapid. For groups exposed to constant temperature there was an increase in oxygen consumption ( M _accl, mg kg⁻¹ h⁻¹) with increasing temperature ( T ), the relationship being approximated by an exponential model: M _accl= 46·53e0·^{086 T} . At 14·3° C_fluc oxygen consumption declined during the 3–4 days following the temperature shift, but remained higher than at 14·4° C_const. This indicates that small temperature fluctuations have some additional influences that increase metabolic rate. Ammonia excretion rates showed diel variations. Excretion was lower at 11° C_const than at other temperatures, and increases in temperature had a significant effect on ammonia excretion rate. Fluctuating (14·3° C_fluc) temperature did not influence ammonia excretion relative to constant temperature (14·4° C_const).     

Effect of rising temperature on the heat resistance of Listeria monocytogenes in meat emulsion

The heat resistance of a strain of L. monocytogenes was determined both in broth and in meat emulsion. The D -values for meat emulsion were approximately two to three times higher than those for broth and also the z -value increased significantly. The micro-organism proved to be more resistant when the cells were heated up slowly (0·5°C/min) to constant temperatures of 60, 63 and 66°C in meat emulsion. The D ₆₀, D ₆₃ and D ₆₆ were, respectively 12·95, 5·4 and 2·3 min. Results may have implications in the survival of Listeria monocytogenes in particular food preparations.     

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 Effect of Water Activity on the Growth and Respiration of Pseudomonas fluorescens

The growth rate of Pseudomonas fluorescens was greater and continued at lower water activity ( a _w) values when glycerol controlled the a _w of glucose minimal medium than when the a _w was controlled by NaCl and sucrose. Growth was not observed below 0·945, 0·970 and 0·964 a _w when glycerol, sucrose and NaCl respectively controlled the a _w. The catabolism of glucose, Na lactate and DL-arginine as measured by respirometry was completely inhibited at a _w values greater than the minimum for growth when the a _w was controlled with NaCl. When the a _w was controlled with glycerol, catabolism of the three substrates continued at a _w values significantly below the a _w for growth on glucose. Catabolism of glucose in the presence of sucrose occurred at a level below the minimum growth a _w but catabolism of the other two substrates ceased at a _w values greater than the minimum growth a _w. Arrhenius plots between 10° and 34°C of the growth rate in glucose minimal medium at 0·98 a _w showed that the order of inhibition was sucrose > NaCl > glycerol. The order of inhibition differed when Arrhenius plots of catabolism of glucose was examined between 10° and 34 °C, namely NaCl > sucrose > glycerol. The mechanism of action of solutes controlling a _w is discussed.     

Biochemical characterization of pectate lyases produced by fluorescent pseudomonads associated with spoilage of fresh fruits and vegetables

An improved method for purification of pectate lyases (PLI and PLII) from culture fluids of Pseudomonas fluorescens CY091 and Ps. viridiflava PJ-08-6 by using a phosphocellulose cation exchanger was described. Analysis of purified PLI and PLII by sodium dodecyl sulphate-polyacrylamide and isoelectric focusing gel electrophoresis revealed that both enzymes had been purified to near homogeneity. Optimal Ca²⁺ concentration required for PLI and PLII activity was determined to be 0·5 mmol l⁻¹. The Ca²⁺ requirement could not be replaced by other metal cations such as Mg²⁺, Cu²⁺, Zn²⁺, Fe³⁺ and Co²⁺. Optimal pH for activity was determined to be between 8·5 and 9·0. The K _m values for sodium polygalacturonate were 1·28 and 1·11 mg ml⁻¹ for PLI and PLII, respectively. Both PLI and PLII were stable at low temperatures (25°C or below) for at least 1 month. However, at 37°C, the activity decreased 50% in 36 h. Optimal temperatures for activity were estimated to be 46° and 52°C for PLI and PLII, respectively. Thermal stability of both enzymes at elevated temperatures (48°C or higher) increased when CaCl₂ or a positively charged molecule such as polylysine was present, but decreased when polygalacturonate or a negatively charged molecule such as heparin was present. PLI and PLII exhibit differential degrees of sensitivity to group-specific inhibitors, including iodoacetic acid and diethylpyrocarbonate. This result suggests that both sulphydryl and imidazole groups are important for the catalytic function of PLI and PLII.     

Influence of culture conditions on exopolysaccharide production by Lactobacillus rhamnosus strain C83

The influence of culture conditions on growth and exopolysaccharide (EPS) production by Lactobacillus rhamnosus strain C83 was investigated using fermentor batch cultures. A temperature shift (from 37 to 25 °C) at the beginning of the exponential growth phase (0–5 h) enhanced EPS production. Furthermore, an optimal environmental pH of 6·2–7·2 improved the performance of strain C83 and partially anaerobic culture conditions (pO₂ from 0 to 10%) triggered EPS over-production by the cells. The sugar composition of this polymer was independent of culture conditions, such as carbon source, medium composition, temperature, pH, pO₂ and growth phases. In all cases, galactose and glucose were the principal components.     

Growth, diet and metabolism of common wolf–fish in the North Sea, a fast–growing population

The von Bertalanffy growth parameters for common wolf–fish Anarhichas lupus in the North Sea were: male: L ∞=111·2 cm, t ₀=–0·43 and K =0·12; and female: L ∞=115·1 cm, t ₀=–0·39 and K =0·11, making this the fastest growing stock reported. Resting metabolic rates (RMR±S.E.) and maximum metabolic rates (MMR±S.E.) for six adult common wolf–fish (mean weight, 1·39 kg) at 5° C were 12·18±1·6 mg O₂ kg^–1 h^–1 and 70·65±7·63 mg O₂ kg^–1 h^–1 respectively, and at 10° C were 25·43±1·31 mg O₂ kg^–1 h^–1 and 113·84±16·26 mg O₂ kg^–1 h^–1. Absolute metabolic scope was 53% greater at 10° C than at 5° C. The diet was dominated by Decapoda (39% overall by relative occurrence), Bivalvia (20%) and Gastropoda (12%). Sea urchins, typically of low energy value, occupied only 7% of the diet. The fast growth probably resulted from summer temperatures approximating to the optimum for food processing and growth, but may have been influenced by diet, and reduced competition following high fishing intensity.     

Effects of temperature, body size and feeding on rates of metabolism in young‐of‐the‐year haddock

The mean rate of oxygen consumption (routine respiration rate, R _R, mg O₂ fish⁻¹ h⁻¹), measured for individual or small groups of haddock Melanogrammus aeglefinus (3–12 cm standard length, L _S) maintained for 5 days within flow‐through respiratory chambers at four different temperatures, increased with increasing dry mass ( M _D). The relationship between R _R and M _D was allometric ( R _R = α  M ^b ) with b values of 0·631, 0·606, 0·655 and 0·650 at 5·0, 8·0, 12·0 and 15·0° C, respectively. The effect of temperature ( T ) and M _D on mean R _R was described by     indicating a Q ₁₀ of 2·27 between 5 and 15° C. Juvenile haddock routine metabolic scope, calculated as the ratio of the mean of highest and lowest deciles of R _R measured in each chamber, significantly decreased with temperature such that the routine scope at 15° C was half that at 5° C. The cost of feeding ( R _SDA) was c . 3% of consumed food energy, a value half that found for larger gadoid juveniles and adults.     

The Resistance of Spores of Clostridium botulinum Type E to Heat and Radiation

Spores of Clostridium botulinum type E have been screened for resistance to heating in aqueous suspension at 60, 70 and 80°. D ¹⁷⁶ ranged from <0·33 min to 1·25 min. z values of eight strains were between 9·0 and 10·7, and for one strain was 7·4.
Radiation survivor curves have been plotted for nine strains, and a further nine strains screened. The "shoulder" extends to about 0·4 megarad, whereafter the curve becomes exponential within a D value from 0·065 to 0·16 megarad.     

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).