The role of development and adrenal steroids in the regulation of the mineralocorticoid receptor messenger RNA.

The ontogeny, adrenal-feedback regulation and regional distribution of the mineralocorticoid receptor (MR) mRNA were examined in the rat brain and kidney. In the kidney, MR mRNA levels in the adult were only 25-30% of the neonatal concentration. Adrenalectomy caused a 35% increase in total brain MR mRNA and a 94% increase in kidney MR mRNA levels. Examination of the regional distribution of the MR mRNA within the brain revealed that the hippocampus had the highest levels, and the mRNA abundance increased after adrenalectomy. The administration of dexamethasone to intact animals resulted in a significant reduction of MR mRNA in the kidney of neonatal rats but not in the brain. These data indicate that there are developmental changes in MR gene expression in kidney and that adrenal steroids can modulate MR gene expression in both the brain and kidney.

Developmental and hormonal regulation of glucocorticoid receptor messenger RNA in the rat.

The biological action of glucocorticoids is dependent upon tissue-specific levels of the glucocorticoid receptor (GR). During stress, the hypothalamic-pituitary-adrenal axis is stimulated, and high levels of glucocorticoids circulate. This axis is modulated by negative feedback by glucocorticoids, which inhibit hypothalamic and pituitary hormone secretion and downregulate GR gene expression. To study the developmental tissue-specific regulation of the GR, we measured the relative concentration of GR mRNA in fetal, neonatal, adult, and aged rats and examined the effects of dexamethasone on GR gene expression. Three different tissue-specific developmental patterns of GR mRNA accumulation were found. In addition, there was an age-dependent tissue-specific pattern in the feedback regulation of GR mRNA by glucocorticoids. In the fetus and neonate, GR mRNA abundance was not regulated by circulating glucocorticoids. The adult pattern of glucocorticoid feedback inhibition of GR mRNA expression appeared between 2 and 7 d of life in liver, and after 7 but before 14 d of age in brain. The GR was biologically active in the 2-d-old neonate, however, since dexamethasone enhanced gene expression of angiotensinogen, which is another glucocorticoid responsive gene. These data demonstrate that the GR gene is regulated by both developmental and tissue-specific factors, and provide another molecular basis for ontogenic variations in the hypothalamic-pituitary-adrenal response to stress.

Thermal inactivation of Listeria monocytogenes within bovine milk phagocytes.

Thermal resistance of intracellular and freely suspended Listeria monocytogenes that was associated with a milkborne outbreak of listeriosis was studied by using the sealed tube and slug flow heat exchanger methods. Test temperatures for the former method were 57.8, 62.8, 66.1, and 68.9 degrees C (136, 145, 151, and 156 degrees F, respectively); whereas those for the latter method were 66.1, 68.9, 71.7, and 74.4 degrees C (151, 156, 161, and 166 degrees F, respectively). The heating menstruum was sterile, whole milk. The intracellular inoculum was generated from an in vitro phagocytosis reaction by using endotoxin-induced bovine milk phagocytes. The phagocyte population consisted of 88% neutrophils, 8% macrophages, and 4% lymphocytes. Neutrophils harbored the majority of intracellular L. monocytogenes. The mean level of infectivity in the phagocyte population was 43%, and there were 26.1 +/- 19.3 bacteria per cell (10(4) viable cells per ml of test milk). Initial bacterial counts for the freely suspended and intracellular experiments (the latter was based on a sonically disrupted sample) were 10(6) L. monocytogenes cells per ml. Heat-stressed bacteria were recovered by direct plating in parallel with recovery from an enrichment broth; both methods gave comparable results. The predicted D62.8 degrees C (145 degrees F) value for intracellular sealed tube studies was 53.8 s (ZD = 5.6 degrees C [10.0 degrees F]), indicating a safe 33.4 D margin of inactivation for vat pasteurization (62.8 degrees C for 30 min).(ABSTRACT TRUNCATED AT 250 WORDS)

Thermal Resistance of Disease-Associated Salmonella typhimurium in Milk.

The thermal resistance of Salmonella typhimurium cultures that had been associated with a major milkborne oubreak of salmonellosis was determined in raw whole milk. Thirteen patient stool isolates and 24 implicated pasteurized milk isolates at concentrations of 1 × 105/ml were screened for heat resistance at 51.8°C. A representative milk strain was heated in replicate at four temperatures from 51.8 to 68.3°C. The zD value was calculated to be 5.3°C. Mean D-value estimates at 51.8°C were 24.0 and 22.8 min for patient and milk isolates, respectively. Extrapolated D71.7°C values were 0.24 and 0.22 s, and did not differ significantly (α = 0.05). These isolates would not survive proper pasteurization.

Thermal Resistance of Listeria monocytogenes in Dairy Products.

The thermal resistance of Listeria monocytogenes strain Scott A that had been associated with a recent milkborne outbreak of listeriosis was determined in whole and skim milk, heavy cream, and ice cream mix. L. monocytogenes suspended at concentrations of approximately 1 × 105 cells/ml was heated at temperatures ranging from 52.2 to 79.4°C at various contact times. The D71.7°C values computed for milk samples ranged from 0.9 to 2.7 s. The D7.94°C value in ice cream mix was 0.5 s. The zD value for fluid products ranged from 5.8 to 7.1°C; the zF value for ice cream mix was 7.0°C. The L. monocytogenes suspensions would not survive a proper pasteurization process given to raw dairy products.

Thermal resistance of intracellular Listeria monocytogenes cells suspended in raw bovine milk.

The thermal resistance of Listeria monocytogenes associated with a milk-borne outbreak of listeriosis was determined in parallel experiments by using freely suspended bacteria and bacteria internalized by phagocytes. The latter inoculum was generated by an in vitro phagocytosis reaction with immune-antigen-elicited murine peritoneal phagocytes. The heat suspension medium was raw whole bovine milk. Both suspensions were heated at temperatures ranging from 52.2 to 71.7 degrees C for various periods of time. Mean D values for each temperature and condition of heated suspension revealed no significant differences. The extrapolated D71.7 degrees C (161 degrees F) value for bacteria internalized by phagocytes was 1.9 s. Combined tube and slug-flow heat exchanger results yielded an estimated D71.7 degrees C value of 1.6 s for freely suspended bacteria. The intracellular position did not protect L. monocytogenes from thermal inactivation.

Gas chromatographic detection of D-(-)-2,3-butanediol and butyric acid produced by sporeformers in cream-style corn and canned beef noodle soup: collaborative study.

A gas chromatographic method that identifies sporeformers as the cause of spoilage in swollen cans of low-acid foods was collaboratively studied in 2 stages. Two organic compounds produced by sporeformers, D-(-)-2,3-butanediol and butyric acid, are measured in the upper phase after centrifugation of the liquid portion of the can contents. Each sample is assayed on 2 packed columns designed for the assay of aqueous solutions of volatile fatty acids, using flame ionization detectors. For study 1, 16 duplicate inoculated cans of cream-style corn and beef noodle soup were sent to 9 collaborators. For study 2, 7 collaborators received 11 duplicate inoculated cans of the 2 foods. Duplicate uninoculated cans of each food served as negative controls. The inocula were 6 sporeforming organisms (4 Clostridium and 2 gas-forming Bacillus species) and 2 nonsporeformers. After the deletion of marginal samples, the percentages of correctly identified sporeformers and nonsporeformers in beef noodle soup were 83 (110/132) and 90 (54/60), respectively; corresponding percentages for cream-style corn were 80 (98/123) and 100 (35/35). The method has been adopted official first action.

Thermal Resistance of Listeria monocytogenes in Milk.

The thermal resistance of Listeria monocytogenes associated with a milkborne outbreak of listeriosis was determined in buffer and whole milk. Thermal resistance was stable over a 2-year period and could not be altered by selecting heat-stressed survivors. The rate of inactivation was linear and did not differ significantly between pH 5.5 and 9.0. When portions of whole milk containing 1 × 105 cells of L. monocytogenes /ml were heated at seven temperatures from 52.2 to 74.4°C, the D-values ranged from 1683.7 to 0.7 s, respectively. The zD-value was 6.3°C. The D-value at 71.7°C was 0.9 s. L. monocytogenes would not survive the pasteurization process.

Thermal inactivation of Yersinia enterocolitica in milk.

Three strains of Yersinia enterocolitica isolated from milk had D values at 62.8 degrees C from 0.24 to 0.96 min and z values of 5.11 to 5.78 degrees C. Since the pasteurization processes for dairy products recommended by the Food and Drug Administration are adequate to destroy large concentrations of these organisms, Y. enterocolitica in pasteurized milk probably results from substandard processing or recontamination after pasteurization.

Heat Processing of Oysters Naturally Contaminated with Vibrio cholerae Serotype 01.

Pathogenic Vibrio cholerae 0-Group 1 survived for more than 3 weeks in artificial sea water with little loss in viability. Live oysters placed in such contaminated, artificial sea water took up but did not concentrate V. cholerae . Heat treatments provided by an in-can pasteurization process and by preparation of naturally contaminated oysters according to common recipes effectively reduced the numbers of V. cholerae by 5 logs/g.

Thermal inactivation of ileal loop-reactive Clostridium perfringens type A strains in phosphate buffer and beef gravy.

The thermal resistance of spore crops produced from each of two ileal loop-reactive strains of Clostridium perfringens type A was determined in two suspending vehicles consisting of 0.067 M (pH 7.0) phosphate buffer and a commercial beef gravy. D115.6 values obtained in buffer and enumerated after pretreatment with sodium ethylenediaminetetraacetate and recovery in plating medium containing lysozyme were two- to threefold greater than those obtained without this treatment. D115.6 values obtained with beef gravy were less than those obtained in buffer with or without lysozyme; however, the D98.9 and D104.4 values were 1.3 to 2 times greater than those obtained in buffer with lysozyme. The z values were within the ranges reported by previous investigators.

Heat resistance of ileal loop reactive Bacillus cereus strains isolated from commercially canned food.

Sporeformers isolated from a commercially canned food were identified as Bacillus cereus, lactose-positive variants. The thermal resistance of spore crops produced from each of two representative cultures was determined in 0.067 M phosphate buffer at pH 7.0. The D121.1 values for one isolate were approximately 0.03 min (z = 9.9C), whereas the D121.1 values for the other isolate were 2.35 min (z = 7.9 C). Thermal inactivation results for heat-stressed isolates from each strain showed no significant alteration in heat resistance from that of the two parent spore crops. Both isolates were reactive when injected into the ligated rabbit ileum.

Survival of Vibrio parahaemolyticus in cooked seafood at refrigeration temperatures.

The growth and survival of two strains of Vibrio parahaemolyticus isolated during food-borne gastroenteritis outbreaks in Japan and surface inoculated on cooked shrimp, shrimp with sauce, or cooked crab were tested at various refrigeration temperatures during a 48-h holding period. On cooked shrimp and crab, the vibrios grew well at 18.3 C, but their numbers declined gradually at 10 C and below. At 12.8 C, vibrios remained static for the most part. Thus, it appeared that 12.8 C was the borderline temperature for growth of the organism on cooked seafood. When cocktail sauce was added to surface-inoculated shrimp at a ratio of 2:1, the vibrio die-off rate was accelerated. In the shrimp and sauce few cells remained after 48 h, but in the sauce alone die-off was complete at 6 h.

Detection of sulfa drugs and antibiotics in milk.

A disc assay method for testing sulfa drugs and antibiotics in milk was developed wherein Bacillus megaterium ATCC 9855 was used as the test organism and Mueller-Hinton agar was used as the test substrate. Incubation was at 37 C for 4 to 5 hr. The test procedure is an improvement over the Bacillus subtilis-Antibiotic Medium No. 1 method, as described in Standard Methods for the Examination of Dairy Products, in that it is sensitive to eight sulfa drugs and to bacitracin without a significant change in sensitivity to eight other antibiotics commonly used for mastitis therapy.

Thermal resistance of salmonellae isolated from dry milk.

Salmonella anatum, S. binza, S. cubana, S. meleagridis, S. newbrunswick, and S. tennessee isolated from dry milk, and S. senftenberg 775W were studied for heat resistance to determine whether these organisms would survive pasteurization as recommended by the 1965 Pasteurized Milk Ordinance of the U.S. Public Health Service. Thermal inactivation determinations were made on washed cells of the test microorganisms suspended in sterile whole milk. Excluding S. senftenberg, D values ranged from 3.6 to 5.7 sec at 62.8 C, from 1.1 to 1.8 sec at 65.6 C, and from 0.28 to 0.52 sec at 68.3 C. Corresponding values for S. senftenberg were 34.0, 10.0, 1.2, and 0.55 sec for respective exposure temperatures of 65.5, 68.3, 71.7, and 73.9 C. The present milk pasteurization processes as recommended by the Public Health Service will inactivate all seven strains of salmonellae studied, provided that the initial concentration does not exceed a calculated 3 x 10(12) salmonellae per ml of milk.