Comparison of Direct Plating Media for the Isolation and Enumeration of Enterococci in Certain Frozen Foods

A total of 15 agar media were examined for their yield, selectivity, readability, and simplicity of preparation and use. A thallium medium of Barnes was selected as the better of the high yield-fair selectivity type of medium and an azide-citrate medium of Reinbold appeared to be the better of the low yield-high selectivity type of medium. Sodium carbonate (optimal concentration, 0.20%) was found to increase recovery substantially when added to certain media, especially in the presence of 0.05% Tween 80. When these two ingredients were incorporated into a medium modified after Slanetz and Bartley, the resultant medium was superior to other media for the isolation and enumeration of enterococci in certain frozen foods, such as peas and hamburger, by the direct plating method.     

Improved Procedure for Identification of Group D Enterococci with Two New Media

The aerosol survival in air was determined for Pasteurella tularensis live vaccine strain (LVS) as a function of relative humidity (RH). Three different preparations of bacteria were used: (i) liquid suspension of P. tularensis LVS in spent culture medium; (ii) powders of P. tularensis LVS freeze-dried in spent culture fluid; (iii) P. tularensis LVS freeze-dried in spent culture fluid and then reconstituted with distilled water and disseminated as a liquid suspension. Preparation (i) gave greatest survival at high RH and lowest survival at intermediate RH. Preparation (ii), in contrast, gave greatest survival at low RH and minimum survival at 81% RH. Preparation (iii) was the same as preparation (i), i.e., the process of freeze-drying and reconstituting with distilled water before aerosol formation had little or no effect upon aerosol survival as a function of RH. Hence, control of aerosol survival appears to be through the water content of P. tularensis LVS at the moment of aerosol generation rather than the water content of the bacteria in the aerosol phase.     

Numerical Taxonomy of Catalase-negative Cocci Isolated from Frozen Vegetables

Taxonomic data for 135 cultures of bacteria isolated from frozen snap beans and whole kernel corn were analyzed by computer. Of the isolates, 79% were grouped into five major clusters. Two of the groups consisted of leuconostocs, whereas the other three contained homofermentative types. The intergroup similarities suggested a generic level of similarity. Feature frequencies for the groups were calculated, and properties of the different groups are presented. Many of the isolates differed from described species.     

Comparison of Several Laboratory Media for Presumptive Identification of Enterococci and Group D Streptococci

Bile-esculin (Difco), modified bile-esculin (Difco), selective enterococcus (Pfizer Co.), and eosin-methylene blue agar media were evaluated for accuracy in identifying group D streptococci. The regular and modified bile-esculin media performed equally well, but the selective enterococcus and eosin-methylene blue agars did not accurately differentiate the group D from non-group D streptococci. A modified 6.5% NaCl broth was compared with unmodified 6.5% NaCl broth and Streptococcus faecalis (SF; Difco) broth for accuracy in differentiating enterococci from non-enterococci. The modified and unmodified broths worked equally well in the salt tolerance test, but the lot-to-lot variability of SF broth made this medium unusable as an indicator for enterococci. With all seven media, the number of strains giving positive tests decreased when the tests were incubated at 45 C as compared with 35 C, and the number of strains giving negative tests increased. Thus, the number of false-positive identifications decreased, but the number of false-negative identifications increased. Variability in the susceptibility of group D non-enterococcal streptococci to oxacillin and methicillin sensitivity disks limited the usefulness of these tests for presumptive identification of either enterococci or group D streptococci.     

Detection and Enumeration of Fecal Indicator Organisms in Frozen Sea Foods: II. Enterococci

Consistently high recoveries of enterococci as compared to the low numbers of coliforms obtained from the same samples of frozen sea foods are indirect evidence that enterococci are better indicators of contamination in such foods.

The use of azide dextrose broth, modified by the incorporation of bromthymol blue, and of ethyl violet azide broth as presumptive and confirmation tests, respectively, were found to be highly specific for the detection and enumeration of enterococci in these samples. Tetrazolium agar medium, when used as a third step after the confirmation test, provides a reliable differentiation of Streplococcus faecalis types from other group D streptococci. A simple procedure is described for further identification of S. faecalis varieties and other enterococcal species.

Incidence of biotypes within certain species is noted and relationships of these subgroups to the organisms described by other workers is discussed.

The striking resistance of all group D streptococci to dihydrostreptomycin and polymyxin B seems to offer promise for evolving a new selective medium for these organisms.

     

Cutting Unfixed Frozen Sections for Fluorescence Antibody Studies

A simplified method of section cutting, dispensing with guide attachment on microtome blade, and suitable for serial sections of unfixed frozen tissue of 4μ is described. Essential features are low temperature maintenance (—13°C), critical angle of knife and moistening of slides in cold alcohol or isopentane. This method has been found suitable for fluorescence antibody studies where maintenance of low temperatures is necessary.     

Studies on Regeneration Media for Bacillus subtilis Protoplasts

Bacillus subtilis protoplasts regenerate on media containing horse serum, bovine serum or gelatin. These compounds could be replaced by polyvinyl pyrrolidone or dextran, and a medium which contained 30 g polyvinyl pyrrolidone and 20 mg casamino acids per liter with chemically defined ingredients was especially useful for selection of prototrophs, e.g., by protoplast fusion. Polyvinyl pyrrolidone and other plasma expanders stimulated protoplast division in liquid media and improved protoplast survival on agar media.     

Numerical Taxonomy of Gram-Positive and Catalase-Positive Rods Isolated from Frozen Vegetables

One hundred isolates from peas, beans, and corn were compared with cultures of Corynebacterium, Microbacterium, and Arthrobacter by use of numerical taxonomic procedures. Six groups (clusters), representing 75% of the isolates, resembled Corynebacteriaceae. There was some doubt regarding the genera represented because the groups were not closely related to many of the known strains. The relationship of the different groups to each other, as well as a number of their properties, is presented.     

无公害蔬菜营养与施肥研究进展

本文综述了无公害蔬菜定义、卫生评价标准及生产环境评价标准。对无公害蔬菜生产中常用的氮肥在土壤中转化、种类和形态、用量和时期与硝酸盐关系,过量氮肥对蔬菜体内硝酸盐累积影响,对地下水质污染和人体健康的影响,磷肥过量引起流失造成水体富营养化和磷肥中镉对土壤生态环境污染的影响,以及畜禽粪肥、城市垃圾和污泥堆肥对蔬菜品质及环境影响进行了评述。对近二十年来土壤栽培条件下应用于无公害蔬菜营养诊断与平衡施肥研究的     

无公害蔬菜营养与施肥研究进展

本文综述了无公害蔬菜定义、卫生评价标准及生产环境评价标准。对无公害蔬菜生产中常用的氮肥在土壤中转化、种类和形态、用量和时期与硝酸盐关系,过量氮肥对蔬菜体内硝酸盐累积影响,对地下水质污染和人体健康的影响,磷肥过量引起流失造成水体富营养化和磷肥中镉对土壤生态环境污染的影响,以及畜禽粪肥、城市垃圾和污泥堆肥对蔬菜品质及环境影响进行了评述。对近二十年来土壤栽培条件下应用于无公害蔬菜营养诊断与平衡施肥研究的养分平衡法、肥料效应函数法、NPK比例法、土壤测试指数法、植物营养外观及测试指标诊断法、DRIS作物营养诊断施肥综合法,植株硝酸盐速测反射仪法等进行了综述。     

湛江常见野菜安全可食性探讨

对湛江野菜所含的硝酸盐、维生素C、胡萝卜素及氨基酸进行了测定和分析。结果表明,刺苋和野当归富含维生素C和胡萝卜素,硝酸盐含量较低,实为品质上乘野蔬。东风菜和枸杞虽然胡萝卜素含量很高,但前者维生素C和氨基酸含量较低,后者硝酸盐含量已超过三级蔬菜标准。不宜大量食用。其余几种野菜营养成分比传统蔬菜高,属于安全食用性野菜。     

Enterococci in Insects

Enterococci were obtained from 213 of 403 insects cultured during a 14-month period, in numbers from 10(3) to 3 x 10(7)/g of insect. Insects were taken only from nonurban, wild, and cultivated fields and woods. In species of insects carrying them, enterococci were not always present in every individual cultured, and often more than one species of enterococcus occurred within a species. Enterococci were obtained from certain insects taken in the field during the dormant season, suggesting their role as overwintering agents. They were generally present in species feeding on nectar, succulent plant parts, and on and ir forest litter, but not from insects feeding on less succulent leaves and stems. Streptococcus faecalis was recovered from 32%, Streptococcus faecium from 22.4%, and Streptococcus faecium var. casseliflavus from 43.5% of members of the 37 taxa of insects. S. faecalis and S. faecium var. casseliflavus exhibit a high percent of conformity to the properties published for them. The heterogeneity in properties of S. faecium is similar to that found for the species taken from plants. Many fail to grow in broth at 45 C or in broth containing 6.5% NaCl; 50% of the cultures ferment both melezitose and melibiose, and a few ferment neither sugar. The remainder ferment melibiose only. Failure to reduce methylene blue in milk by S. faecalis and S. faecium is correlated with the inability to ferment lactose. More than 93% of the cultures of S. faecalis digest casein in milk from the top downward, following the production of a soft, flowing curd. Because this property is not characteristic of S. faecalis taken from humans, the reaction in litmus milk is suggested as a means of differentiation between cultures of remote and innocent origin in nature and recent, human pollution.     

Enterococci in Insects

Enterococci were obtained from 213 of 403 insects cultured during a 14-month period, in numbers from 10³ to 3 × 10⁷/g of insect. Insects were taken only from nonurban, wild, and cultivated fields and woods. In species of insects carrying them, enterococci were not always present in every individual cultured, and often more than one species of enterococcus occurred within a species. Enterococci were obtained from certain insects taken in the field during the dormant season, suggesting their role as overwintering agents. They were generally present in species feeding on nectar, succulent plant parts, and on and ir forest litter, but not from insects feeding on less succulent leaves and stems. Streptococcus faecalis was recovered from 32%, Streptococcus faecium from 22.4%, and Streptococcus faecium var. casseliflavus from 43.5% of members of the 37 taxa of insects. S. faecalis and S. faecium var. casseliflavus exhibit a high percent of conformity to the properties published for them. The heterogeneity in properties of S. faecium is similar to that found for the species taken from plants. Many fail to grow in broth at 45 C or in broth containing 6.5% NaCl; 50% of the cultures ferment both melezitose and melibiose, and a few ferment neither sugar. The remainder ferment melibiose only. Failure to reduce methylene blue in milk by S. faecalis and S. faecium is correlated with the inability to ferment lactose. More than 93% of the cultures of S. faecalis digest casein in milk from the top downward, following the production of a soft, flowing curd. Because this property is not characteristic of S. faecalis taken from humans, the reaction in litmus milk is suggested as a means of differentiation between cultures of remote and innocent origin in nature and recent, human pollution.