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A Anellis

Publications and source records attributed to A Anellis.

16 recordsLinked to original sources

Comparative dose-survival curves of representative Clostridium botulinum type F spores with type A and B spores.

Radiation survival data of proteolytic (Walls 8G-F) and non-proteolytic (Eklund 83F) type F spores of Clostridium botulinum were compared with dose-response data of radiation-resistant type A (33A) and B (40B) spores. Strain Eklund 83F was as resistant as strain 33A, whereas strain Walls 8G-F was the most sensitive of the four strains tested. The methods suggested for computing both an initial shoulder and a D value for the dose-survival curves yielded results comparable to the graphic techniques used to obtain these two parameters.

Cesium Radioisotopes↗

Cryogenic gamma irradiation of prototype pork and chicken and antagonistic effect between Clostridium botulinum types A and B.

Inoculated, irradiated pork (2,300 cans) and chicken (2,000 cans) pack studies were performed to establish the 12D dose for these foods. Each can was inoculated with a mixture of 10(6) spores of each of 10 strains of Clostridium botulinum (five type A and five type B), or a total of 10(7) spores. The cans received a series of increasing doses of gamma rays (60Co) at -30 +/- 10 degrees C; they were incubated for 6 months at 30 +/- 2 degrees C and examined for swelling, toxicity, and recoverable botulinal cells. The highest rate of swelling for both foods occurred within the first week of incubation, and maximum swelling was observed within 4 to 5 weeks. The minimal experimental sterilizing dose (ESD) based on flat, nontoxic sterile cans was 3.0 less than ESD less than or equal to 3.2 Mrad for pork and 4.0 less than ESD less than or equal to 4.2 Mrad for chicken. An analysis of the partial spoilage data by extreme-value statistics indicated with 90% confidence that the rate of spore death in the two foods was not a normal distribution, but appeared to favor a shifted exponential function. Based on the latter distribution, and assuming one most resistant strain in the mixture of 10 used, the 12D dose computed to 4.37 Mrad, with a shoulder of 0.11 Mrad, for pork and to 4.27 Mrad, with a shoulder of 0.51 Mrad, for chicken. An assumption that there were two or more most resistant strains in the inoculum progressively lowered the 12D dose. There was an apparent antagonism between the irradiated type A and B viable strains in the two foods. Cans with type B cells and toxin predominated over cans with type A cells and toxin, but cans with a mixture of type A and B toxins predominated over cans with a mixture of Type A and B cells. At the highest sublethal doses, only type A cells survived in pork, but in chicken there was a least one type B strain that was at least as resistant as type A strains.

Animals↗

Influence of postirradiation incubation temperature on recovery of radiation-injured Clostridium botulinum 62A spores.

The number of colonies formed by unirradiated Clostridium botulinum 62A spores was independent of temperature, in the range from 20 to 45 degrees C (in 5 degrees C increments); no colonies developed at 50 degrees C. Spores irradiated at 1.2 or 1.4 Mrads produced more macrocolonies at 40 degrees C than at higher or lower temperatures. Apparently, radiation-injured spores were capable of repair of 40 degrees C than at the other temperatures studied. More than 99% of the radiation (1.2 Mrads) survivors were injured and were unable to form macrocolonies in the presence of 5% NaCl. The germinated radiation-injured spores were also sensitive to dilution, resulting in the loss of viability of 77 to 79% of the radiation survivors. At 30 and 40 degrees C, the irradiated spores did not differ significantly in the extent of germination (greater than 99% at both 30 and 40 degrees C), emergence (64% at 30 degrees C and 67% at 40 degrees C), and the maximum number of emerged cells that started to elongate (69% at 30 degrees C and 79% at 40 degrees C). However, elongation was remarkably more extensive at 40 degrees C than at 30 degrees C. Many elongated cells lysed within 48 h at 30 degrees C, indicating an impaired repair mechanism. If the radiation-injured spores were incubated at 40 degrees C in the recovery (repair) medium for 8 to 10 h, they germinated, emerged, and elongated extensively and were capable of repair. If, after 8 to 10 h at 40 degrees C, these cultures were shifted to 30 degrees C, the recovery at 30 increased by more than eightfold, resulting in similar colony counts at 30 and 40 degrees C. Thus, repair appeared to be associated with outgrowth. Repair did not occur in the presence of chloramphenicol at 40 degrees C, whereas penicillin had no effect, suggesting that the repair involved protein synthesis but did not require multiplication.

Chloramphenicol↗

Radiation resistance of spores of some Clostridium perfringens strains.

Clostridium perfringens spores (eight strains) were irradiated in a model system with 60Co gamma rays at -30 C. The quantal response data obtained were analyzed with extreme value statistics. It was found (at the 95% confidence level) that all eight strains followed the same rate of death and that this rate was probably (at the 95% level) not exponential. The statistics did not exclude, however, a normal, lognormal, Weibull, or related rate of spore kill. A more definitive study would be required to distinguish between the latter distributions.

Clostridium perfringens↗

Low-temperature irradiation of beef and methods of evaluation of radappertization process.

An inoculated, irradiated beef pack (1,240 cans) was conducted for the determination of microbiological safety for unrestricted human consumption. Each can contained a mixture of 10(6) spores of each of 10 strains of Clostridium botulinum (5 type A and 5 type B), or a total of 10(7) spores/can. The cans were irradiated to various doses (100 cans/dose) with 60Co gamma rays at -30 +/- 10 C, incubated at 30 +/- 2 C for 6 months, and examined for swelling, toxicity, and recoverable botulinal cells. The minimal experimental sterilizing dose based on nonswollen, nontoxic sterile cans were 2.2 less than experimental sterilizing dose based on nonswollen, nontoxic sterile cans was 2.2 less than experimental sterilizing dose less than or equal to 2.6 Mrad. Using recoverable cells as the most stringent criterion of spoilage, and assuming the conventional simple exponential (without an initial shoulder) rate of spore kill, the "12D" dose was 3.7 Mrad when estimated on the basis of mixture of 10 strains totaling 10(7) spores/can, and 4.3 Mrad if it is assumed that each can of beef contained 10(6) spores of a single most resistant strain and all of these spores were of identical resistances. However, an analysis of the data by extreme value statistics indicated with 90% confidence that the spore death rate was not a simple exponential but might be a shifted exponential (with an initial shoulder), Weibull, lognormal, or normal, with a "12D" equivalent of about 3.0 Mrad regardless of the initial spore density per can. There was an apparent antagonism between the irradiated type A and B strains in the cans. Some of the cans contained type B toxin but did not include type B viable cells. Other cans had a mixture of type A and B toxins, but a large number of these cans did not yield recoverable type B cells. However, type A viable cells could always be demonstrated in those cans containing type A toxin.

Animals↗

Evaluation of media, time and temperature of incubation, and method of enumeration of several strains fo Clostridium perfringens spores.

Two basal media, containing the ingredients found in common in both SPS (BBL) and TSN (BBL) media and in the previously described media of Schaedler et al. (1965) and Starr et al (1971), but minus antibiotics, were selected as the most suitable for the enumeration of Clostridium perfringens spores in a model system. These media were also used to study the influence of the presence of glucose, xylose, or ribose in various concentrations (0, 0.01, 0.1, and 1.0%) on colony morphology and spore recovery. As the sugar concentration in the basal agar medium increased, the colonies of all the test organisms also increased in size, and more of the black colonies became white in color. At the 1.0% sugar level, glucose permitted only white colony development, whereas the pentoses were completely inhibitory. Both pour plates and most-probable-number tubes were inoculated with the spores of several strains of C. perfringens and incubated at 20, 30, 37, and 45 C for 24, 48, and 72 h. Statistical analyses of the enumeration data indicated, at the 99% confidence level, that a Trypticase(BBL)-yeast extract-glucose-sulfite-iron agar gave maximal population estimates at 37 C in 72 h.

Agar↗

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.

Bacteria↗

Production of types A and B spores of Clostridium botulinum by the biphasic method: effect on spore population, radiation resistance, and toxigenicity.

Spores of three strains each of type A and type B Clostridium botulinum were produced both by a biphasic (solid medium overlaid with an aqueous phase) and by a "conventional" (deep broth culture) procedure. Sporogenesis by the biphasic system was more rapid, convenient, and economical, and yielded as many or more heat-resistant (80 C, 10 min) spores per milliliter as by the conventional technique. Of several aqueous phases [thiamine-hydrochloride, yeast extract, (NH(4))(2)SO(4)] tested with strain 62A, the highest spore colony counts were obtained with 2.0% (NH(4))(2)SO(4). The six strains formed maximum spore numbers in 5 to 6 days of incubation. Spores produced by the two methods had essentially equal radiation resistances (D and lag values), and their subcultures gave similar toxin titers (LD(50) values).

Ammonium Sulfate↗

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 </= 3.0, 2.0 < ESD </= 2.5, and 1.5 < ESD </= 2.0 Mrad, in that order. Assuming exponential spore death, the 12D values, or minimal radiation doses (MRD), were 3.24, 2.44, and 2.65 Mrad, respectively. Estimation of the MRD values by a method which assumes that spore death in the cans follows a normal distribution, yielded 3.09, 2.57, and 2.39 Mrad, respectively. Weibull analyis of the pooled 10-strain viable cell spoilage data of the screening packs for codfish cake or corned beef suggested that spore death in the cans follows a normal distribution yielded 3.09, 2.57, pooled data were not amenable to such analysis. Sublethal doses (0.5, 0.75 Mrad) increased the visible spoilage rate of corned beef over that of unirradiated controls. Apparently radiation-injured spores of C. botulinum were sensitized to the presence of food additives such as curing salts, NaCl, and spices.

Animals↗

Clostridium perfringens. I. Sporulation in a biphasic glucose-ion-exchange resin medium.

A biphasic culture medium suitable for cultivation and sporulation of Clostridium perfringens, C. botulinum, and C. sporogenes was devised. The medium designed for use in a disposable, compartmented, plastic film container contained peptones, yeast extract, minerals, an anion exchange resin, and glucose in 4% agar as the solid phase and (NH(4))(2)SO(4) and 0.1% agar as the liquid phase. With the biphasic system, it was not necessary to use active cultures as inocula. Growth was at least equal to that obtained in conventional media, and spore production of 9 out of 12 strains of C. perfringens equalled or usually exceeded that of conventional media.

Agar↗

Radiation sterilization of prototype military foods. 3. 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 </= 3.0 Mrad, and based on non-spoiled sterile cans was 3.5 < ESD </= 4.0 Mrad. The theoretical minimal radiation dose (MRD), the 12D equivalent, varied with the method of computation: 4.74, 4.33 +/- 0.17, and 4.19 to 4.99 Mrad were obtained by the Weibull, Spearman-Karber, and Schmidt-Nank techniques, respectively. Calculation of D and MRD values by the conventional Schmidt-Nank method produced increasing values with rising dosage; this finding was compared with the data derived by the other two methods of calculation. Suggestions for estimating the MRD of a prototype radiation process are offered.

Animals↗

Estimation of radiation resistance values of microorganisms in food products.

Several statistical methods, including the conventional technique of Schmidt and Nank, were evaluated for estimating radiation resistance values of various strains of Clostridium botulinum by the use of partial spoilage data from an inoculated ham pack study. Procedures based on quantal response were preferred. The tedious but rigorous probit maximum likelihood determination was used as a standard of comparison. Weibull's graphical treatment was the method of choice because it is simple to utilize, it is mathematically sound, and its ld(50) values agreed closely with the reference standard. In addition, it offers a means for analyzing the type of microbial death kinetics that occur in the pack (exponential, normal, log normal, or mixed distributions), and it predicts the probability of microbial death with any radiation dose used, as well as the dose needed to destroy any given number of organisms, without the need to assume the death pattern of the partial spoilage data. The Weibull analysis indicated a normal type kinetics of death for C. botulinum spores in irradiated cured ham rather than an exponential order of death, as assumed by the Schmidt-Nank formula. The Weibull 12D equivalent of a radiation process, or the minimal radiation dose (MRD), for cured ham was consistently higher than both the experimental sterilizing dose (ESD) and the Schmidt-Nank average MRD. The latter calculation was lower than the ESD in three of the five instances examined, which seems unrealistic. The Spearman-Kärber estimate was favored as the arithmetic technique on the bases of ease of computation, close agreement with the reference method, and providing confidence limits for the ld(50) values.

Clostridium botulinum↗

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 </= 2.5 Mrad, and based on non-spoiled sterile cans was 3.0 < ESD </= 3.5 Mrad (the latter was supported by the computed theoretical 12D dose); (iii) D values calculated from botulinal survival data indicated that, as a group, the type A strains were more radioresistant than type B strains; strains 12885A and 41B, with respective D values of 0.242 and 0.175, represented the most resistant of each type; (iv) swollen cans did not always contain toxin, nor were toxic cans always swollen; (v) viable C. botulinum can exist for 6 months at 30 C without producing visible or toxic spoilage at doses of 3.0 Mrad and lower, including, in some instances, 0.0 Mrad; and (vi) a phenomenon similar to heat activation of spores occurred at sublethal radiation doses.

Animals↗

Incidence of mesophilic Clostridium spores in raw pork, beef, and chicken in processing plants in the United States and Canada.

The anaerobic film pouch technique was used to quantitate and isolate clostridial spores in 2,358 samples of raw meat (1,078 of chicken, 624 of beef, 656 of pork). Of 19,727 putrefactive anaerobic (PA) sporeformers isolated, 1 was confirmed by mouse protection testing to be Clostridium botulinum type C. This isolate was obtained from a Western Canada chicken sample which contained 5.33 clostridia per gram. These data indicate a very low incidence of botulinal contamination in raw meats at the packing-plant level (0.042% of 2,358 samples) and an almost 20,000:1 ratio of nonbotulinal PA sporeformers to mesophilic C. botulinum spores. The mean level of PA contamination was 2.8 PA sporeformers per gram of meat; 77% of the samples contained three or less PA sporeformers per gram. Small but statistically significant differences in the incidence of clostridial spores were noted for season, geographical region, and type of meat.

Animals↗