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Volatiles produced by microorganisms isolated from refrigerated chicken at spoilage.

Volatile components present at spoilage of refrigerated chicken breasts were identified using high-vacuum-low-temperature distillation techniques followed by analysis with combined temperature-programmed gas chromatography and mass spectrometry. A comparison was made of the compounds detected from both irradiated and non-irradiated muscle stored at 2 and 10 degrees C under both aerobic and anaerobic conditions. Isolates were randomly selected from the spoiled poultry, identified, and evaluated for their ability to produce volatile spoilage noted when grown on radiation-sterilized chicken. Several isolates that produced off-odors on sterile chicken breasts were examined. Twenty-two compounds were associated with spoilage. Some of the compounds found on both irradiated and unirradiated samples were considered to play only a minor role in the spoilage aroma or were present in low concentrations, since the aroma of spoiled irradiated chicken lacked the harsh odor notes typical of spoiled unirradiated chicken. Fifteen of the 22 compounds were considered to be unique to unirradiated, aerobically spoiled samples. Nine of these compounds, hydrogen sulfide, methyl mercaptan, dimethyl sulfide, dimethyl disulfide, methyl acetate, ethyl acetate, heptadiene, methanol, and ethanol, were found on chicken spoiled at both 2 and 10 degrees C. xylene, benzaldehyde, and 2,3-dithiahexane were detected only in samples stored at 2 degrees C and methyl thiolacetate, 2-butanone, and ethyl propionate were associated with 10 degrees C spoilage. Fifty-eight isolates randomly selected from fresh, radiation-pasteurized, and unirradiated spoiled poultry were classified taxonomically, and 10 of them, which produced spoilage odors on sterilized chicken breasts, were selected for subsequent analysis of their volatiles. Isolates identified as Pseudomonas putrefaciens and Pseudomonas species that were members of groups I and II of Shewan's classification, as well as Flavobacterium and oxidative Moraxella, produced a number of the compounds found in the aroma of spoiled chicken. A total of 17 compounds were identified. Whereas no isolate produced all of the aroma compounds found in the aroma of spoiled chicken, together they did produce the nine found in unirradiated samples spoiled at either 2 or 10 degrees C, as well as methyl thiolacetate and xylene. Six compounds were present in the volatiles produced by the isolates but were absent in the volatiles identified from spoiled chicken. These were hydrogen cyanide, methyl isopropyl sulfide, 2-propane thiol, methyl propionate, ethyl benzene, and an unidentified compound.

Aerobiosis↗

Efficacy of media and methods for detecting and enumerating Campylobacter jejuni in refrigerated chicken meat.

A study was undertaken to compare several enrichment and direct isolation media for their suitability to detect and enumerate five strains of Campylobacter jejuni in refrigerated (5 degrees C) chicken meat. The influence of CO2 on survival at 5 degrees C was also investigated. Selective enrichment media evaluated included Preston broth (PB), selective semisolid brucella medium (SSBM), Campylobacter enrichment broth (CEB), VTP brucella-FBP broth (VTP), Rosef and Kapperud Campylobacter enrichment broth (RKCEB), and Doyle and Roman enrichment broth (DREB). Direct isolation agars included Campy brucella agar (CBAP), blood-free Campylobacter medium (BFCM) and modified Butzler agar (MBA). Comminuted chicken meat was inoculated with C. jejuni, sealed under atmospheric gas or CO2, and stored at 5 degrees C for up to 21 days. Viable population was determined by the most-probable-number technique (PB, SSBM, CEB, VTP, and RKCEB, followed by plating on CBAP, BFCM, and MBA), enrichment on DREB, followed by plating on CBAP, BFCM, and MBA, and direct isolation on CBAP, BFCM, and MBA. Without exception, direct plating of samples was superior to the most-probable-number technique for enumerating C. jejuni; MBA was inferior to CBAP and BFCM, and DREB performed at least as well as other enrichment media evaluated. Carbon dioxide afforded protection against death of three of the five strains of C. jejuni tested.

Animals↗

Bacteriocin-mediated inhibition of Clostridium botulinum spores by lactic acid bacteria at refrigeration and abuse temperatures.

The bacteriocinogenicity of Lactococcus lactis ATCC 11454, Pediococcus pentosaceus ATCC 43200, P. pentosaceus ATCC 43201, Lactobacillus plantarum BN, L. plantarum LB592, L. plantarum LB75, and Lactobacillus acidophilus N2 against Clostridium botulinum spores at 4, 10, 15, and 35 degrees C was investigated by modified deferred and simultaneous antagonism methods. All the strains, except L. acidophilus N2, produced inhibition zones on lawns of C. botulinum spores at 30 degrees C. L. plantarum BN, L. lactis ATCC 11454, and P. pentosaceus ATCC 43200 and 43201 were bacteriocinogenic at 4, 10, and 15 degrees C. Supplementation of brain heart infusion agar with 0 to 5% NaCl increased the radii of inhibition zones during simultaneous antagonism assays. Detectable bacteriocin activities were extracted from freeze-thawed agar cultures of L. plantarum BN and L. lactis ATCC 11454 which had been grown at 4 and 10 degrees C. These results suggest that low levels of L. plantarum BN or L. lactis ATCC 11454, in the presence of 3 or 4% NaCl, could be formulated into minimally processed refrigerated food products for protection against possible botulism hazards.

Bacteriocins↗

Inhibitory effect of combinations of heat treatment, pH, and sodium chloride on a growth from spores of nonproteolytic Clostridium botulinum at refrigeration temperature.

Nonproteolytic strains of Clostridium botulinum will grow at refrigeration temperatures and thus pose a potential hazard in minimally processed foods. Spores of types B, E, and F strains were used to inoculate an anaerobic meat medium. The effects of various combinations of pH, NaCl concentration, addition of lysozyme, heat treatment (85 to 95 degrees C), and incubation temperature (5 to 16 degrees C) on time until growth were determined. No growth occurred after spores were heated at 95 degrees C, but lysozyme improved recovery from spores heated at 85 and 90 degrees C.

Botulinum Toxins↗

A pigment-producing spoilage bacterium responsible for violet discoloration of refrigerated market milk and cream.

A psychrophilic strain of bacteria identified as Chromobacterium lividum was established as the causative agent of an outbreak of violet discoloration in refrigerated, pasteurized retail milk and cream. The organism was rod-shaped, gram-negative, and produced viscid colonies with abundant violet pigment on Tryptone glucose yeast extract agar. Growth was abundant at 4 C but none occurred at 37 C. Growth in milk was characterized by a dark violet ring at the surface after a few days, and the deep violet color gradually extended through the product in older cultures. Some proteolysis occurred. The pigment appeared to be similar to that of other known species of Chromobacterium and assisted in identification of the genus of the causative organism. The isolated strain of C. lividum was destroyed by exposure to 56 C for 5 min which suggested postpasteurization contamination as the source of the spoilage organism in commercial milk and cream.

Animals↗

Effect of refrigeration on the antinematodal efficacy of ivermectin.

Several observations have suggested that the anthelmintic ivermectin can affect nematodes by non-oral entry into the nematode body. To investigate this possibility further, we refrigerated Caenorhabditis elegans at 5 C to prevent its locomotion and to block the pharyngeal pumping that is so prominent a feature of its feeding. Worms were exposed to ivermectin (1-25 microg/ml) at that temperature for 1 hr, after which the medium was replaced by unmedicated medium at room temperature. After 1 hr at room temperature the worms were examined and counted to determine the degree to which irreversible immobilization had occurred. The drug was significantly less effective at 5 C than at room temperature. This reduction in potency could be attributed to a general cold-induced decline in the rate of the biochemical processes involved in drug action. Alternatively, the reduction could be attributed to the cold-induced blockade of pharyngeal pumping, which would suggest that the efficacy of ivermectin is partially the result of oral intake of drug. The fact that antinematodal efficacy was not entirely abrogated and reached a significant level despite blockade of pharyngeal pumping supports the former interpretation and is in accord with earlier indications that ivermectin can enter by non-oral routes. This conclusion is further supported by the observation that ivermectin is active against the nonfeeding third-stage larva of Haemonchus contortus.

Animals↗

Functionality and physico-chemical characteristics of bovine and caprine mozzarella cheeses during refrigerated storage.

Low moisture part-skim Mozzarella cheeses (MC) were manufactured using fresh bovine and caprine milk to study melting, physico-chemical, textural, and microstructural properties of the cheeses during 8 wk of refrigerated storage. Structural changes in cheese matrix were evaluated by scanning electron microscopy and by proteolytic patterns using nitrogen solubility, SDS-PAGE, and Gel-pro analyzer. Meltability of ripened cow and goat MC were not different when fat content of both milks were standardized, whereas bovine MC formed a significantly larger amount of free oil throughout the experiment. The results of the proteolytic patterns, texture attribute (cohesiveness), and microstructure revealed that bovine MC had a greater structural degradation of cheese matrix than caprine MC during the storage. Elevated protein degradation in bovine MC led to more intense brown color formation than the goat counterpart when the cheeses were baked. The melting characteristics showed high positive correlation (r = 0.51 to 0.80) with proteolysis, whereas it was negatively correlated with textural characteristics. Among textural attributes, cohesiveness was highly inversely correlated with melting characteristics (r = -0.69 to -0.88). High negative correlations were also observed between proteolytic parameters and textural attributes (r = -0.48 to -0.81).

Animals↗

Viability of Lactobacillus acidophilus and Lactobacillus casei in fermented milk products during refrigerated storage.

The viability was investigated of five strains of Lactobacillus acidophilus and one strain of Lactobacillus casei that were added as adjuncts to yogurt and cultured buttermilk during 28 d of refrigerated storage at 5 to 7 degrees C. A modification of LBS (Lactobacillus selection) agar was used for the enumeration of L. acidophilus and L. casei. The medium allowed the colony formation of the adjunct bacteria while preventing colony formation of the traditional yogurt or buttermilk starter cultures. At each sampling period, colonies from the selective agar medium were isolated for confirmation of identity to confirm that only L. acidophilus and L. casei were enumerated, that their characteristics did not change during storage, or both. The strains of L. acidophilus varied in both cultured products. In buttermilk, L. acidophilus MUH-41, O-16, and L-1 exhibited no significant loss in viability, but strains 43121 and La-5 did. No significant loss in viability of L. acidophilus MUH-41 and L-1 occurred in yogurt prepared using culture CM2; however, strains 43121, O-16, and La-5 lost viability. In the yogurt prepared using culture YC-4, L. acidophilus 43121 exhibited no significant loss in viability, but MUH-41, O-16, L-1, and La-5 did. There was no loss in viability of L. casei GG during storage of any of the cultured products.

Animals↗

High salt lysates: a simple method to store blood samples without refrigeration for subsequent use with DNA probes.

Blood specimens to be tested for the presence of Plasmodium falciparum using specific DNA probes can be stored as high salt lysates (HSL) without refrigeration. The lysates are prepared from 100 microliter blood samples by a simple 3-step procedure using 2 volumes of H2O to lyse the erythrocytes (step I), 1 volume of a detergent/EDTA mix to lyse the parasites (step II), followed by the addition of 1 volume cesium trifluoroacetate (CsTFA) (step III). The parasite DNA was found to be undegraded, as shown by the unaltered pattern of repetitive sequences obtained after storage of up to 1 month at 37 degrees C, due to the inhibition of DNA degrading enzymes by the cesium salt. The bulk of protein can be removed from the samples by a 1-step precipitation. The addition of 0.3 volumes of a mixture of ethanol: chloroform: isoamyl alcohol (2.5:1:0.04 v/v) precipitates greater than 90% of the proteins from the lysates, leaving greater than 86% of the parasite DNA in the supernatant. The reduced protein content of the samples, when applied to solid supports, results in an increased signal: background ratio on autoradiograms.

Animals↗

Enumeration and identification of yeasts associated with commercial poultry processing and spoilage of refrigerated broiler carcasses.

Yeasts associated with broiler carcasses taken from various stages of commercial poultry processing operations and broiler carcasses stored at refrigerated temperatures were enumerated and identified. Whole carcass rinses were performed to recover yeasts from carcasses taken from a processing facility and processed carcasses stored at 4 degrees C for up to 14 days. Yeasts in the carcass rinsates were enumerated on acidified potato dextrose agar and identified with the MIDI Sherlock Microbial Identification System. Dendrograms of fatty acid profiles of yeast were prepared to determine the degree of relatedness of the yeast isolates. Findings indicated that as the carcasses are moved through the processing line, significant decreases in the number of yeasts associated with broiler carcasses usually occur, and the composition of the yeast flora of the carcasses is altered. Significant (P < 0.05) increases in the yeast population of the carcasses generally occur during storage at 4 degrees C, however. Furthermore, it was determined that the same strain of yeast may be recovered from different carcasses at different points in the processing line and that the same strain of yeast may be isolated from carcasses processed on different days in the same processing facility.

Animals↗

Incidence of Clostridium perfringens in commercially produced cured raw meat product mixtures and behavior in cooked products during chilling and refrigerated storage.

A total of 445 whole-muscle and ground or emulsified raw pork, beef, and chicken product mixtures acquired from industry sources were monitored over a 10-month period for vegetative and spore forms of Clostridium perfringens. Black colonies that formed on Shahidi-Ferguson perfringens (SFP) agar after 24 h at 37 degrees C were considered presumptive positive. Samples that were positive after a 15-min heat shock at 75 degrees C were considered presumptive positive for spores. Of 194 cured whole-muscle samples, 1.6% were positive; spores were not detected from those samples. Populations of vegetative cells did not exceed 1.70 log10 CFU/g and averaged 1.56 log10 CFU/g. Of 152 cured ground or emulsified samples, 48.7% were positive, and 5.3% were positive for spores. Populations of vegetative cells did not exceed 2.72 log10 CFU/g and averaged 1.98 log10 CFU/g; spores did not exceed 2.00 log10 CFU/g and averaged 1.56 log10 CFU/g. Raw bologna (70% chicken), chunked ham with emulsion, and whole-muscle ham product mixtures were inoculated with C. perfringens spores (ATCC 12916, ATCC 3624, FD1041, and two product isolates) to ca. 3.0 log10 CFU/g before being subjected either to thermal processes mimicking cooking and chilling regimes determined by in-plant temperature probing or to cooking and extended chilling regimes. Populations of C. perfringens were recovered on SFP from each product at the peak cook temperatures, at 54.4, 26.7, and 7.2 degrees C, and after up to 14 days of storage under vacuum at 4.4 degrees C. In each product, populations remained relatively unchanged during chilling from 54.4 to 7.2 degrees C and declined slightly during refrigerated storage. These findings indicate processed meat products cured with sodium nitrite are not at risk for the growth of C. perfringens during extended chilling and cold storage.

Animals↗

Recovery rate of Listeria monocytogenes from commercially prepared frankfurters during extended refrigerated storage.

To assess the prevalence of Listeria monocytogenes in vacuum-sealed packages of frankfurters, about 33,000 packages (1 lb each) were obtained by a third-party contractor from 12 volunteer commercial manufacturers over a 2-year period. The 12 producers, each of which contributed about 2,700 packages of frankfurters from one production run, comprised 9 large and 3 small plants located in eight U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) districts in 10 states. Five days after manufacture, 500 packages were sampled at the USDA/Agricultural Research Service (ARS) Eastern Regional Research Center (ERRC) in Wyndmoor, Pa., by the USDA/ARS package rinse method. At regular intervals during subsequent storage at 4 and 10 degrees C, an additional 200 packages were tested for the pathogen at each sampling point. From a statistical perspective, L. monocytogenes was not recovered from any of the products of nine of the producers, whereas the pathogen was recovered at rates of 1.5% (plant 367), 2.2% (plant 439), and 16% (plant 133) from the products of the remaining three plants. In total, 532 of 32,800 (1.6%) packages of frankfurters tested positive for the pathogen. The recovery rates did not change appreciably over time, there was no appreciable difference in L. monocytogenes recovery rates with respect to frankfurter storage temperature (4 or 10 degrees C), and the seasonality of manufacture had no influence on recovery rate. Molecular subtyping of multiple L. monocytogenes-positive isolates from each plant revealed that profile A (serotype 1/2a) was displayed by about 90% of the 1,105 isolates tested. However, in some cases it was also possible to recover more than one profile from a given plant. This study provides estimates of the prevalence, types, and viability of L. monocytogenes associated with commercially prepared frankfurters during extended refrigerated storage.

Animals↗

Effectiveness of trisodium phosphate treatment against pathogenic and spoilage bacteria on poultry during refrigerated storage.

To determine the efficacy of trisodium phosphate (TSP) against pathogenic and spoilage bacteria on poultry and to assess the influence of the bacterial combination of inoculum on TSP effect, chicken legs were coinoculated with similar concentrations of a pathogenic (Salmonella enterica ser. Enteritidis or Listeria monocytogenes) and a spoilage (Pseudomonas fluorescens or Brochothrix thermosphacta) bacteria. Samples were dipped in TSP (12%, 15 min) or were not treated (control). Microbiological analysis and pH determinations were carried out at 0, 1, 3, and 5 days of storage (3 degrees C). Significant bacterial reductions (marked in gram-negative species) were observed on TSP-treated samples throughout refrigerated storage. Inoculum composition scarcely influenced the TSP effect against gram-positive bacteria. However, greater reductions were observed on gram-negative bacteria (Salmonella Enteritidis and P. fluorescens) when samples were coinoculated with B. thermosphacta and L. monocytogenes, respectively. Values of pH were higher in TSP-treated than in control samples at all sampling times. The combination of bacteria in inocula did not have a significant influence on pH values.

Animals↗

Fates of seeded Escherichia coli O157:H7 and Salmonella on selected fresh culinary herbs during refrigerated storage.

The fates of seeded Escherichia coli O157:H7 and Salmonella on selected fresh culinary herbs were evaluated at a refrigerated temperature (4 degrees C). Fresh herbs, including cilantro, oregano, basil, chive, parsley, and rosemary, were inoculated with six-strain mixtures of E. coli O157:H7 and Salmonella, and the microbial populations were monitored at 1, 5, 11, 16, 19, and 24 days. For both pathogens, a significant decrease in the population (P < 0.0001) occurred within the first 5 days of storage (< 0.8 log). Both pathogens remained the highest on cilantro and the lowest on rosemary (P < 0.0001). Storage time had a significant effect on the survival of E. coli O157:H7; populations declined as storage time progressed. Although storage of cilantro, basil, and chive was terminated after 19 days because of deteriorated quality, significant numbers of both pathogens were recovered from the remaining fresh herbs after 24 days of storage. The results showed that both bacteria were extremely persistent on all test herbs under the test conditions. The results also reinforce the concept that, once contaminated, bacterial pathogens can persist on fresh herbs throughout a normal distribution time.

Chive↗

[Suitability study for super sweet corn on the ears hybrids under refrigerated conditions evaluating chemicals microbiological and sensorial characteristics].

Krispy King, Víctor and 324, super sweet hybrids (sh2) were cultivated in San Joaquín, estado Carabobo, Venezuela. The scheme was stablished to produce refrigerated fresh ears to be commercialized. The chemistry, microbiology and sensorial characteristics were evaluated at 0; 7; 14; 21 and 28 days of storage. One hundred ears of each hybrid were picked at the ripe fresh stage and packed in polystyrene trays covered with polyethylene. The storage temperature was 4 degrees C +/- 1 degree C. The scheme used was well adapted, allowing a good stability of the ears until 28 days of storing. The plastic cover avoid the lost of humidity. The soluble solids, total sugars and pH went down during the storage. The acidity and the microorganisms increased as expected. The sensorial variables kept the same for Krispy king and Víctor, while the hybrid 324 shown the lowest humidity content, the highest count of microorganisms and the poorest sensorial quality.

Chimera↗

Stability of ranitidine in intravenous admixtures stored frozen, refrigerated, and at room temperature.

The stability of ranitidine in concentrations of 0.5, 1.0, and 2.0 mg/mL in admixtures with commonly used i.v. fluids was studied. The admixture vehicles were 0.9% sodium chloride, 5% dextrose, 10% dextrose, 5% dextrose and 0.45% sodium chloride, and 5% dextrose with lactated Ringer's (DLR) injections in polyvinyl chloride bags. Three bags were prepared for each test solution and stored under each of the following conditions: seven days at room temperature (23 +/- 1 degrees C) in normal laboratory lighting, 30 days at 4 degrees C, and 60 days at -20 degrees C followed by either seven days at room temperature (in light) or 14 days at 4 degrees C. Ranitidine content was determined by high-performance liquid chromatography at several intervals. Color, clarity, and pH were also examined. Ranitidine concentrations remained greater than or equal to 90% of initial concentrations under all storage conditions except in the frozen DLR admixtures. Drug loss in the DLR admixtures was greatest at the lower ranitidine concentrations. The only visual changes were yellow color in the thawed DLR admixtures and those containing ranitidine 2.0 mg/mL in 5% dextrose and 0.45% sodium chloride. Slight increases in the pH of some admixtures were noted. Ranitidine is stable for seven days at room temperature and 30 days at 4 degrees C at all concentrations and in all vehicles studied. At the studied concentrations, the drug is stable in admixtures frozen for 60 days and stored for seven days at room temperature or 14 days refrigerated, except in DLR admixtures; these admixtures should not be stored frozen.

Drug Combinations↗

[Contact renal refrigeration with continuous control of the intrarenal temperature in surgery for staghorn calculi].

Surgical treatment of staghorn calculi is a meticulous, difficult, and time-consuming procedure which requires the taking of contact films during operation. Incision of the renal parenchyma is frequently also necessary. To limit blood loss, and mainly in order to have a clear field of view, intrarenal blood circulation has to be interrupted by clamping the renal pedicle. This circulatory arrest can be supported for only 20 minutes, without major risks to the renal parenchyma, under normal temperature conditions. However, at least in the authors' hands, more than 10 minutes are necessary between the taking of the contact film and its return after development to the operation room. In contrast, lowering renal temperature by 10 to 20 degrees C enables ischemia to be maintained for up to 4 hours without adverse consequences. Contact refrigeration of the kidney by surrounding it with crushed ice is used, permanent control of temperature being maintained by a thermocouple linked to a continuous digital display. This simple, easy to install method does not require high investments or sophisticated material. It has enabled continuous clamping to be applied for periods of up to nearly 2 hours, with normal follow-up I.V.P. one week later.

Humans↗