Comparative Resistance of Nonsporogenic Bacteria to Low-Temperature Gamma Irradiation

A total of 36 microorganisms, comprising 19 species of 11 genera, were screened for radiation resistance with (60)Co gamma rays at a radiation temperatore of -80 +/- 2 C in phosphate buffer (pH 7.0) under vacuum. Micrococcus radiodurans was the most resistant organism. An initial population of 2.8 x 10(5) cells per dose of this species survived 2.4 but not 2.7 Mrad. Of the remaining 18 species with initial populations of about 10(6) cells per dose, Streptococcus faecium survived 0.9 to 1.5 Mrad, depending on the strain tested. S. faecalis QM survived 0.9 but not 1.2 Mrad. S. faecalis 1539 and Alcaligenes faecallis survived 0.6 but not 0.9 Mrad. Three species of Salmonella, one strain each of Escherichia coli, Streptococcus lactis, and Aerobacter aerogenes survived 0.3 but not 0.6 Mrad. The remaining 22 bacteria did not survive 0.3 Mrad, the lowest dose tested. Detailed survival curve determinations for four strains of S. faecium, the most resistant of the test bacteria of public health significance, indicated the following order of resistance at -80 C: alpha21 > theta12 = F(6) > FEC. Each strain produced two exponential survival curves with different slopes, the breaks occurring at 0.3 to 0.5 Mrad. The D values (doses which reduce the microbial population by 90%) of the more resistant cell fractions were two- to three-fold higher than the more sensitive cell fraction. The resistance of strain alpha21 was determined at different radiation temperature (+5, -30, -80, -140, -196 C). The D value-radiation temperature relationship followed a quadratic equation. Computations of E(a) and Q(10) values (activation energy and temperature coefficient, respectively) showed a very small thermodynamic effect on radiation death. An Arrhenius evaluation of the temperature effect on cell kill indicated that there was no simple physicochemical mechanism which might explain the change in D value as a function of temperature.     

The rationale and a computer evaluation of a gamma irradiation sterilization dose determination method for medical devices using a substerilization incremental dose sterility test protocol

The experimental procedure described is designed to allow calculation of the radiation sterilization dose for medical devices to any desired standard of sterility assurance. The procedure makes use of the results of a series of sterility tests on device samples exposed to doses of radiation from 0.2 to 1.8 Mrad in 0.2 Mrad increments. From the sterility test data a 10^-2 sterility level dose is determined. A formula is described that allows a value called DS Mrad to be calculated. This is an estimate of the effective radiation resistance of the heterogeneous microbial population remaining in the tail portion of the inactivation curve at the 10^-2 dose and above. DS Mrad is used as a D ₁₀ value and is applied, in conjunction with the 10^-2 sterility level dose, to an extrapolation factor to estimate a sufficient radiation sterilization dose. A computer simulation of the substerilization process has been carried out. This has allowed an extensive evaluation of the procedure, and the sterilization dose obtained from calculation to be compared with the actual dose required. Good agreement was obtained with most microbial populations examined, but examples of both overdosing and underdosing were found with microbial populations containing a proportion of organisms displaying pronounced shoulder inactivation kinetics. The method allows the radiation sterilization dose to be derived from the natural resistance of the microbial population to gamma sterilization.     

Application of Replica Plating and Computer Analysis for Rapid Identification of Bacteria in Some Foods: II. Analysis of Microbial Flora in Irradiated Dover Sole (Microstomus pacificus)

This investigation was carried out to determine the nature of the microbial flora shifts in dover sole as a result of irradiation and storage at 6 C. The relationship was determined between the microorganisms which initially survive irradiation and those making up the final spoilage flora. A total of 2,723 isolates were examined by use of the replica-plating and computer analysis method. The spoilage of the unirradiated control samples during storage at 6 C was almost entirely due to the growth of Pseudomonas. This group, which occupied 25% of the fresh flora, grew to nearly 100% in 2 days of storage. In contrast, irradiation doses of 0.1, 0.2, 0.3, and 0.4 Mrad favored the growth of Achromobacter and yeasts. The Micrococcus, which survived radiation, did not grow at 6 C. At 0.5 Mrad, spoilage of fish samples was due entirely to the growth of yeasts.     

Effect of γ-Irradiation on the Microflora of Freshwater Fish: I. Microbial Load,Lag Period,and Rate of Growth on Yellow Perch (Perca flavescens) Fillets

Microbial flora were compared in irradiated and nonirradiated yellow perch fillets. These studies included effects of irradiation on the total microbial population, the lag phase, and rate of growth in this freshwater fishery product. The work was conducted concurrently with sensory and chemical evaluation, and constituted part of an investigation designed to evaluate the effect of substerilization doses (0.3 and 0.6 Mrad) of Co⁶⁰ γ rays on the storage life of yellow perch fillets at 1.0 or 6.0 C. In five storage tests, total plate counts prior to irradiation did not exceed 8.7 × 10⁵ per gram of sample; this count was reduced nearly 100% by irradiation with either 0.3 or 0.6 Mrad. Progressively lower maximal bacterial populations and lengthened lag phases were obtained as more radiation was used. The growth rate of the population did not appear to decrease significantly. Microbial data obtained in these studies confirmed the sensory and chemical studies, by indicating that irradiation can significantly extend the refrigerated shelf life of freshwater fish.     

Effect of γ-Irradiation on the Microflora of Freshwater Fish: I. Microbial Load, Lag Period, and Rate of Growth on Yellow Perch (Perca flavescens) Fillets

Microbial flora were compared in irradiated and nonirradiated yellow perch fillets. These studies included effects of irradiation on the total microbial population, the lag phase, and rate of growth in this freshwater fishery product. The work was conducted concurrently with sensory and chemical evaluation, and constituted part of an investigation designed to evaluate the effect of substerilization doses (0.3 and 0.6 Mrad) of Co⁶⁰ γ rays on the storage life of yellow perch fillets at 1.0 or 6.0 C. In five storage tests, total plate counts prior to irradiation did not exceed 8.7 × 10⁵ per gram of sample; this count was reduced nearly 100% by irradiation with either 0.3 or 0.6 Mrad. Progressively lower maximal bacterial populations and lengthened lag phases were obtained as more radiation was used. The growth rate of the population did not appear to decrease significantly. Microbial data obtained in these studies confirmed the sensory and chemical studies, by indicating that irradiation can significantly extend the refrigerated shelf life of freshwater fish.     

Radiation Sterilization of Prototype Military Foods: Low-Temperature Irradiation of Codfish Cake, Corned Beef, and Pork Sausage

"Screening" packs comprising 10 lots each of codfish cake, corned beef, and pork sausage, each lot containing about 10(6) spores of a different strain (five type A and five type B) of Clostridium botulinum per can, were irradiated at -30 +/- 10 C with a series of increasing doses (20 replicate cans/dose) of (60)Co gamma rays. The cans were incubated for 3 months at 30 C and examined for swelling, toxin, and recoverable botulinal cells. Based on the latter criterion of spoilage, median lethal dose (LD(50)) and D values were estimated for each strain in each food. The most resistant strain in codfish cake, corned beef, and pork sausage was, respectively, 53B, 77A, and 41B. There was no clear-cut trend in the comparative order of resistance between the two antigenic types among the three foods. LD(50) values gave essentially the same order of resistances as the D values and may be used interchangeably with the latter for the 10 test organisms. "Clearance" packs consisting of the most resistant strain (about 10(7) spores/can) with its respective food were irradiated with a variety of doses at -30 +/- 10 C, using 100 replicate cans/dose (about 10(9) spores/dose). These packs were incubated for 6 months at 30 C and assayed for the three types of spoilage. Based on recoverable cells, the experimental sterilizing doses (ESD) for codfish cake, corned beef, and pork sausage were 2.5< ESD 相似文献   

Relation between radiation resistance and salt sensitivity of spores of five strains of Clostridium botulinum types A, B, and E.

The NaCl tolerance of different strains of Clostridium botulinum varies over a wide range, and the patterns of NaCl inhibition differ distinctly and characteristically from strain to strain. The more radiation-resistant strains, such as 33A, 62A, and 7272A, are more resistant to NaCl, whereas the more radiation-sensitive strains, such as 51B and 1304E, are more sensitive to NaCl. This rule appears to hold irrespective of whether the spores were unirradiated controls or whether they were radiation damaged prior to exposure to NaCl in the recovery media. The data seem to indicate that radiation doses in the shoulder portion of the radiation survival curves did not noticeably sensitive the spores to NaCl, whereas radiation doses in the exponential-decline portion of the survival curve invariably produced a distinct sensitization. Thus, strains 33A and 62A were not sensitized to NaCl by 0.3 to 0.4 Mrad, i.e., in the shoulder portion of the survival curve. Radiation-sensitive strain 51B, which shows no distinct shoulder in its survival curve, was sensitized to NaCl by 0.1 Mrad, the lowest radiation dose employed in this study. These observations seem to suggest a possible relationship between deoxyribonucleic acid repair capacity and salt tolerance.     

Relation between radiation resistance and salt sensitivity of spores of five strains of Clostridium botulinum types A, B, and E.

The NaCl tolerance of different strains of Clostridium botulinum varies over a wide range, and the patterns of NaCl inhibition differ distinctly and characteristically from strain to strain. The more radiation-resistant strains, such as 33A, 62A, and 7272A, are more resistant to NaCl, whereas the more radiation-sensitive strains, such as 51B and 1304E, are more sensitive to NaCl. This rule appears to hold irrespective of whether the spores were unirradiated controls or whether they were radiation damaged prior to exposure to NaCl in the recovery media. The data seem to indicate that radiation doses in the shoulder portion of the radiation survival curves did not noticeably sensitive the spores to NaCl, whereas radiation doses in the exponential-decline portion of the survival curve invariably produced a distinct sensitization. Thus, strains 33A and 62A were not sensitized to NaCl by 0.3 to 0.4 Mrad, i.e., in the shoulder portion of the survival curve. Radiation-sensitive strain 51B, which shows no distinct shoulder in its survival curve, was sensitized to NaCl by 0.1 Mrad, the lowest radiation dose employed in this study. These observations seem to suggest a possible relationship between deoxyribonucleic acid repair capacity and salt tolerance.     

Gamma processing of Arabic bread for immune system-compromised cancer patients.

Arabic bread prepared from local Saudi flour contained a total of up to 10(4) organisms per g. Most of these were bacterial spores that survived the baking process (1.3 X 10(2) to 3.5 X 10(3] and a small number of yeasts and molds (10 to 40 cells per g). The organisms in Arabic bread appear to be harmless to healthy individuals. However, for immune system-compromised cancer patients and bone marrow transplant recipients, it is prudent to irradiate the bread to reduce microbial contamination. The decimal reduction doses (10% survival) for the most radiation-resistant organisms (spore formers) in bread were 0.11 to 0.15 Mrad. Accordingly, 0.6 Mrad was sufficient to reduce the number of spores in Arabic bread by a factor of 10,000, i.e., to less than 1/g. This treatment constitutes radiation pasteurization (radicidation), and to this extent, provides a margin of microbiological safety. Sensory evaluation by the nine-point hedonic scale showed no detectable loss of organoleptic quality of bread up to 0.6 Mrad, while irradiation to 2.5 Mrad induced unacceptable organoleptic changes.     

Gamma processing of Arabic bread for immune system-compromised cancer patients

Arabic bread prepared from local Saudi flour contained a total of up to 10(4) organisms per g. Most of these were bacterial spores that survived the baking process (1.3 X 10(2) to 3.5 X 10(3] and a small number of yeasts and molds (10 to 40 cells per g). The organisms in Arabic bread appear to be harmless to healthy individuals. However, for immune system-compromised cancer patients and bone marrow transplant recipients, it is prudent to irradiate the bread to reduce microbial contamination. The decimal reduction doses (10% survival) for the most radiation-resistant organisms (spore formers) in bread were 0.11 to 0.15 Mrad. Accordingly, 0.6 Mrad was sufficient to reduce the number of spores in Arabic bread by a factor of 10,000, i.e., to less than 1/g. This treatment constitutes radiation pasteurization (radicidation), and to this extent, provides a margin of microbiological safety. Sensory evaluation by the nine-point hedonic scale showed no detectable loss of organoleptic quality of bread up to 0.6 Mrad, while irradiation to 2.5 Mrad induced unacceptable organoleptic changes.     

Radiation Sterilization of Prototype Military Foods: II. Cured Ham

Ten lots of diced cured ham, packed in cans, were inoculated with approximately 10(6)Clostridium botulinum spores per can. Each lot was seeded with a different strain (five type A and five type B strains). All cans were irradiated to various dose levels with Co(60). Evidence provided by swelling, toxicity, and recoverable C. botulinum with 6,350 cans demonstrated that: (i) 4.5 Mrad was more than adequate as a sterilization dose; (ii) the minimal experimental sterilizing dose (ESD) based on nonswollen nontoxic endpoints was 2.0 < ESD 相似文献   

Effect of Temperature of Liquid Nitrogen on Radiation Resistance of Spores of Clostridium botulinum

An apparatus consisting of a Dewar flask and a relay system controlling the flow of liquid nitrogen permitted the irradiation of samples in tin cans or Pyrex tubes at temperatures ranging from 0 ± 1.5 C to -194 ± 2 C. An inoculated pack comprising 320 cans of ground beef containing 5 × 10⁴ spores of Clostridium botulinum 33A per can (10 cans per radiation dose) was irradiated with Co⁶⁰ at 0 and -196 C. Incubation was carried out at 30 C for 6 months. Approximately 0.9 Mrad more radiation was required to inactivate the spores at -196 C than at 0 C. Cans irradiated at -196 C showed partial spoilage at 3.6 Mrad and no spoilage at 3.9 Mrad; the corresponding spoilage-no spoilage doses at 0 C were 2.7 and 3.0, respectively. The majority of positive cans swelled in 2 to 14 days; occasional swelling occurred as late as 20 days. At progressively higher doses, swelling was delayed proportionally to the radiation dose received. The remaining nonswollen cans had no toxin after 6 months of storage, although occasional cans contained very low numbers of viable spores comprising on the average 0.1% of the original spore inoculum. The D₁₀ values in phosphate buffer were 0.290 Mrad for 0 C and 0.396 Mrad for -196 C; in ground beef, the corresponding D₁₀ values were 0.463 Mrad and 0.680 Mrad, respectively. These D₁₀ values indicate that the lethal effect of γ rays decreased at -196 C as compared with 0 C by 13.5% in phosphate buffer, and by 47% in ground beef.     

Radiation Sterilization of Prototype Military Foods. III. Pork Loin

Ten lots of pork loin, packed in cans, were inoculated with approximately 10(6)Clostridium botulinum spores per can. Each lot was seeded with a different strain; five type A and five type B strains were used. The pack comprised 5,690 cans, including controls, and contained about 10(9) spores per dose. The cans were irradiated with Co(60) in the range of 0 to 5.0 Mrad (0.5 Mrad increments) at 5 to 25 C, incubated for 6 months at 30 C, and examined for swelling, toxicity, and recoverable C. botulinum. The minimal experimental sterilizing dose (ESD) based on nonswollen, nontoxic, but nonsterile end points was 2.5 < ESD 相似文献   

Effect of ionizing radiation of the antigenic composition of typhoid bacteria

Changes in the antigenic composition of typhoid bacteria occurring during the exposure of microbial suspension to different doses of gamma irradiation [Co60] ranging between 0.5 and 3.0 Mrad were studied. Immunoelectrophoresis in agar was used to determine the antigenic composition of different samples of irradiated bacteria. The antigenic composition of bacteria irradiated with doses up to 2.5 Mrad was found to be similar to that of non-irradiated bacteria. Antigens demonstrated by means of Vi, H and O ontisera are preserved in these bacteria. However, all irradiated bacteria in general slightly differ from non-irradiated bacteria; this is manifest in a different configuration and position of the precipitation lines in the cathodic part of the immunophoreograms. The content of the component migrating rapidly towards the cathode, evidently the O antigen in the R form, in the irradiated bacteria increases with the dose of radiation. No new serologically active substances, non-existent in non-irradiated bacteria, were found to appear in the process of irradiation.     

Microbial Flora of Irradiated Dungeness Crabmeat and Pacific Oysters

The microorganisms in Dungeness crabmeat (Cancer magister) and Pacific oysters (Crassostrea gigas) were identified by the replica-plating and computer analysis method. The initial flora of the shellfish and the flora change during storage at 7 C were determined. The microbial flora shifts in both shellfish were also determined after irradiation at 0.1 and 0.4 Mrad and during subsequent storage at 7 C. The Achromobacter species predominated in the initial flora of crabmeat (77.0%). The predominant position of this group increased to 99.2% after 0.1 Mrad and 100% after 0.4 Mrad. A large percentage of Lactobacillus was detected in oysters (55.0%). The Lactobacillus species were the predominant survivors after 0.1 Mrad (92.4%) but the predominant survivors after 0.4 Mrad were Achromobacter species (99.3%).     

Irradiation of Composts Used in the Propagation of Agaricus bisporus

Unpasteurized horse manure composts were given radiation doses of 0.1 and 1.0 Mrad in an attempt to increase mushroom yields by eliminating competing microorganisms. Yields were enhanced by 0.1 Mrad but were lower than those obtained with heat-pasteurized composts. Composts subjected to 1.0 Mrad gave the lowest yields, probably because of the generation of ammonia and other toxic compounds. Only temporary changes in the predominant microflora could be detected as a result of ionizing radiations.     

Effect of γ Irradiation on the Microflora of Freshwater Fish: III. Spoilage Patterns and Extension of Refrigerated Storage Life of Yellow Perch Fillets Irradiated to 0.1 and 0.2 Megarad1

Maximal shelf life was determined and microbial flora were compared for irradiated (0.1 and 0.2 Mrad) and nonirradiated yellow perch fillets stored at 1 C. Shelf life was estimated by organoleptic determinations. Microbiological studies included determination of the effects of irradiation on the total aerobic microbial population, lag phase, and rate of growth. Genera of organisms isolated from fillets through the course of microbial spoilage were identified, and the proteolytic activity of the organisms was determined. Plate counts for fish prior to irradiation showed the presence of approximately 10(6) organisms per g of sample. Irradiation to 0.1 and 0.2 Mrad produced 1.4 and 3 logarithm reductions of the initial count, respectively. Irradiation to 0.1 and 0.2 Mrad approximately doubled the product's shelf life. Organisms initially isolated from the nonirradiated fillets, in order of decreasing number, consisted of Flavobacterium, Micrococcus-Sarcina, Achromobacter-Alcaligenes-Mima, Pseudomonas, Microbacterium, Vibrio, Bacillus, Corynebacterium, Lactobacillus, Brevibacterium, and Aeromonas. By the 6th and 9th days of fillet storage, Pseudomonas and the Achromobacter group were the predominant organisms. All members of the genus Flavobacterium, but not all members of the genus Pseudomonas, were proteolytically active on raw fish juice-agar and skim milk-agar media. The Achromobacter group was found to be nonproteolytic on both media. Residual flora of fillets irradiated to 0.1 and 0.2 Mrad consisted of the Achromobacter group, Lactobacillus, Micrococcus-Sarcina, and Bacillus. Their sequence in predominance, however, varied with dose level. Not all proteolytic bacteria in the fillets were eliminated by 0.1 and 0.2 Mrad; proteolytic Micrococcus-Sarcina survived these treatments.     

Activation of poly(ADP-ribose)-polymerase inhibits apoptosis of thymocytes, caused by short-wave ultraviolet radiation

The effect of (i) aphidicolin, a specific inhibitor of delta- and epsilon-polymerases, and nucleotide excision repair; (ii) 3-aminobenzamide, an inhibitor of poly(ADP-ribose) polymerase and base excision repair; and (iii) actinomycin D and cycloheximide, inhibitors of protein and RNA synthesis, respectively, on the induction of suppression of apoptosis of rat thymocytes by different doses of short-wavelength ultraviolet radiation was studied by flow cytometry. 3-Aminobenzamide suppressed the inhibition of apoptosis induced by the doses of short-wavelength ultraviolet radiation higher than 20 J/m2, increasing the cell death to a maximum. Thus, the inhibition of apoptosis by high short-wavelength ultraviolet radiation doses depends on the status of poly(ADP-ribose) polymerase and is prevented by 3-aminobenzamide. As opposed to 3-aminobenzamide, aphidicolin did not affect the cell death at short-wavelength radiation doses higher than 10 J/m2 but induced the apoptosis of unirradiated cells and cells irradiated with short-wavelength ultraviolet radiation doses lower than 10 J/m2. The inhibitors of protein and RNA synthesis cycloheximide and actinomycin D prevented the induction of apoptosis caused by low and medium doses but did not abolish the apoptosis-inhibiting activity of high doses of short-wavelength ultraviolet radiation.     

Suppression of Spoilage Rates of Bombay Duck (Harpodon nehereus) Homogenates by Gamma Radiation

The spoilage rates of unirradiated and irradiated (0.1 Mrad) Bombay duck (Harpodon nehereus) homogenates stored at 0°C, 10°C and 30°C in terms of organoleptic score (OS) total bacterial count (TBC), trimethylamine content (TMA), and total volatile basic nitrogen (TVBN) followed a linear response with increase in temperature. Unirradiated Bombay duck stored at 0°C were found to spoil faster than those irradiated at 0.1 Mrad; the spoilage rates derived for TBC, TMA, TVBN and OS were 0.42, 0.67, 1.93 and ?0.86 respectively for unirradiated samples as compared with corresponding values of 0.31, 0.55, 1.19 and ?0.19 for the irradiated samples. The spoilage rates of unirradiated Bombay duck stored at 10°C and 30°C measured as a function of OS, TMA and TVBN were consistently higher than their irradiated (0.1 Mrad) counterparts. In the case of irradiated samples, TBC was not a satisfactory index of spoilage.     

降雨对秦皇岛西浴场细菌总数和可培养菌群组成的影响

【目的】研究降雨条件对浴场细菌总数和优势菌群组成的影响。【方法】2014年8月强降雨前后采集秦皇岛西浴场3个站位的海水样品,采用荧光显微镜计数法和平板计数法分别对细菌总数和可培养细菌总数进行计数;对群落结构组成进行分析,并对可培养细菌进行鉴定。【结果】雨前3个站位细菌总数和可培养细菌总数平均值分别为5.6×10~9 CFU/L和8.3×10~7 CFU/L,雨后分别为9.2×109 CFU/L和2.1×10~8 CFU/L。在可培养菌群中,变形菌门(Proteobacteria,雨前占80%,雨后占73%)是主要的微生物类群,其次为拟杆菌门(Bacteroides,雨前占12%,雨后占13%)、厚壁菌门(Firmicutes,雨前占7%,雨后占11%)等;肠杆菌属(Enterobacter spp.,21株)、海杆菌属(Marinobacter spp.,13株)、弓形菌属(Arcobacter spp.,13株)、假单胞菌属(Pseudomonas spp.,10株)、芽孢杆菌属(Bacillus spp.,10株)和弧菌属(Vibrio spp.,6株)为雨前可培养细菌优势属,而雨后可培养细菌优势属为肠杆菌属(22株)、海杆菌属(21株)、芽孢杆菌属(14株)、不动杆菌属(Acinetobacter spp.,11株)、假单胞菌属(9株)和弓形菌属(5株)等。【结论】降雨对细菌总数有显著的影响,同时降雨后浴场微生物群落结构发生了改变。