Tryptone Glucose Yeast Extract Research Articles

Facultative thermophilic microorganisms in potato products in retort packaging

Thermally treated potato products in retort packaging with an extended shelf-life (up to 90 days before opening) at room temperature (20 ± 1 °C) from two Latvian vegetable processing companies (A and B) were analysed in this research. The aim of this research was to determine the presence of thermotolerant microbial contamination in thermally treated potato products in retort packaging, to ensure their safety and suitability for immediate consumption. Samples from three production batches were used. The total plate count was determined, and a microscopic examination of potato product samples was performed. Microscopic examination of microorganisms showed the presence of microbial vegetative forms in potato products from company B, but the standard investigation did not register them; therefore, Tryptone Glucose Yeast Extract Agar (TGYA), TGYA enriched with 1 % starch solution and TGYA enriched with 5 % starch solution were used for potential microorganism isolation. Samples were incubated at 25 °C; 37 °C; 41.5 °C; 50 °C; 55 °C and 60 °C temperatures under aerobic and anaerobic conditions, incubation time 12, 24 and 48 h. Facultative thermophilic Bacillus licheniformis bacteria which produce thermotolerant spores were isolated from thermally treated potato products in retort packaging from company B. The average colony count in company B samples was 1.27´100 CFU/g (SD = 78.25). The presence of microorganisms was not detected in potato products produced at company A, while facultative thermophilic and thermotolerant microorganisms were found in potato products produced at company B. DOI: http://dx.doi.org/10.5755/j01.ct.66.1.12363

Growth characteristics of Lactobacillus brevis KB290 in the presence of bile

Live Lactobacillus brevis KB290 have several probiotic activities, including immune stimulation and modulation of intestinal microbial balance. We investigated the adaptation of L. brevis KB290 to bile as a mechanism of intestinal survival. Strain KB290 was grown for 5 days at 37 °C in tryptone–yeast extract–glucose (TYG) broth supplemented with 0.5% sodium acetate (TYGA) containing 0.15%, 0.3%, or 0.5% bile. Growth was determined by absorbance at 620 nm or by dry weight. Growth was enhanced as the broth's bile concentration increased. Bile-enhanced growth was not observed in TYG broth or with xylose or fructose as the carbon source, although strain KB290 could assimilate these sugars. Compared with cells grown without bile, cells grown with bile had twice the cell yield (dry weight) and higher hydrophobicity, which may improve epithelial adhesion. Metabolite analysis revealed that bile induced more lactate production by glycolysis, thus enhancing growth efficiency. Scanning electron microscopy revealed that cells cultured without bile for 5 days in TYGA broth had a shortened rod shape and showed lysis and aggregation, unlike cells cultured for 1 day; cells grown with bile for 5 days had an intact rod shape and rarely appeared damaged. Cellular material leakage through autolysis was lower in the presence of bile than in its absence. Thus lysis of strain KB290 cells cultured for extended periods was suppressed in the presence of bile. This study provides new role of bile and sodium acetate for retaining an intact cell shape and enhancing cell yield, which are beneficial for intestinal survival.

MLST typing of antimicrobial-resistant Propionibacterium acnes isolates from patients with moderate to severe acne vulgaris

Molecular typing data on antimicrobial-resistant Propionibacterium strains are limited in the literature. We examined antimicrobial resistance profiles and the underlying resistance mechanisms in Propionibacterium spp. isolates recovered from patients with moderate to severe acne vulgaris in Greece. The clonallity of the resistant Propionibacterium acnes isolates was also investigated.Propionibacterium spp. isolates were detected using Tryptone-Yeast Extract-Glucose (TYG) agar plates supplemented with 4% furazolidone. Erythromycin, clindamycin, vancomycin, penicillin, co-trimoxazole, doxycycline, minocycline and ciprofloxacin MICs were determined using the gradient strip method. Erythromycin, clindamycin and tetracycline mechanisms of resistance were determined using PCR and sequencing of the domain V of 23S rRNA and 16S rRNA, as well as the presence of the ermX gene. Typing was performed using the multi locus sequence typing (MLST) methodology.Seventy nine isolates from 76 patients were collected. Twenty-three isolates (29.1%) exhibited resistance to erythromycin and clindamycin, while two additional isolates (2.5%) were resistant only to erythromycin. Resistance to tetracycline was not detected. The underlying molecular mechanisms were point mutations A2059G and A2058G. MLST typing of the P. acnes resistant isolates revealed that lineage type IA1 (ST-1, 3 and 52) prevailed (12/18; 66.7%), whilst lineage type IA2 (ST-2 and 22) accounted for five more isolates (27.8%). Susceptible isolates were more evenly distributed between ST types.Propionibacterium spp. from moderate to severe acne vulgaris in Greece are frequently resistant to erythromycin/clindamycin but not to tetracyclines, mainly due to the point mutations A2059G and A2058G. P. acnes resistant isolates were more clonally related than susceptible ones and belonged to a limited number of MLST types.

Host-centric Proteomics of Stool: A Novel Strategy Focused on intestinal Responses to the Gut Microbiota

The diverse community of microbes that inhabits the human bowel is vitally important to human health. Host-expressed proteins are essential for maintaining this mutualistic relationship and serve as reporters on the status of host-microbiota interaction. Therefore, unbiased and sensitive methods focused on host proteome characterization are needed. Herein we describe a novel method for applying shotgun proteomics to the analysis of feces, focusing on the secreted host proteome. We have conducted the most complete analysis of the extracellular mouse gut proteome to date by employing a gnotobiotic mouse model. Using mice colonized with defined microbial communities of increasing complexity or a complete human microbiota ('humanized'), we show that the complexity of the host stool proteome mirrors the complexity of microbiota composition. We further show that host responses exhibit signatures specific to the different colonization states. We demonstrate feasibility of this approach in human stool samples and provide evidence for a "core" stool proteome as well as personalized host response features. Our method provides a new avenue for noninvasive monitoring of host-microbiota interaction dynamics via host-produced proteins in stool.

Development of “Field Level” Chromogenic Assay for Aflatoxin M1 Detection in Milk

Out of eighteen Bacillus spp. screened for aflatoxin M1 (AFM1) sensitivity Bacillus brevis MTCC 3136 was found most sensitive followed by B. megaterium ATCC 9885 at 0.5 ppb (Codex limit). Bacillus brevis 3136 was found weakly positive for lipase activity while B. megaterium 9885 showed significant acetyl esterase activity. Maximum sporulation was achieved in Arret and Krishbaum sporulation agar (90.63%) followed by tryptone yeast extract broth, tryptone glucose yeast extract broth, minimal media and sucrose salt medium. A field level chromogenic assay for AFM1 in milk has been developed using spores of B. megaterium 9885 grown in Arret and Krishbaum sporulation agar. Spores were added with milk containing germinants (dextrose, sucrose, tryptone) and chromogenic substrate followed by incubation at 37EšC for 75 ± 5 min. The colour change from colourless to sky blue colour within 75 ± 5 min is indication of spore germination followed by release of enzyme and its action on chromogenic substrate. However, no colour change indicates that milk is positive for AFM1 at 0.5 ppb. The developed assay was validated with raw, pasteurized and dried milks with a 96% correlation by comparing with radioimmunoassay i.e., Charm 6602 assay. The reliability of assay has also been checked by spiking AFM1 in milk. The developed assay was also validated by certified reference materials of heavy metals and pesticides for checking the cross reactivity of these with our developed assay. The developed chromogenic assay is cost effective, portable, having no interference of antibiotics, detergents, sanitizers, heavy metals and pesticides. The developed assay can be used in dairy farm and laboratories with fewer resources for AFM1 detection in milk.

Commensal symbiosis between a Lactococcus lactis strain and an Enterococcus mundtii strain increases cell yield in constituted broth

To exert their beneficial effects, probiotics need to survive in the stringent conditions of the gastrointestinal tract. Symbiosis between different bacteria is a potential way of enhancing this survival. In developing new probiotic cultures, we investigated the synergic effect between Enterococcus mundtii IFO 13712 and 7 strains of Lactococcus lactis, many of which are widely used as starter bacteria for making dairy products and have probiotic properties. The growth yield of a mixed culture of L. lactis strain Y and IFO 13712 in de Man, Rogosa, and Sharpe broth was greater than that of a single culture. Supernatant from culture of strain IFO 13712 enhanced the growth of strain Y, but that of strain Y did not enhance the growth of strain IFO 13712. This commensalism phenomenon was confirmed by using a simpler tryptone-yeast extract-glucose (TYG) broth. Increased cell yield in mixed culture of the 2 strains compared with single cultures was observed in TYG broth in the presence of both Tween 80 and citrate but not in TYG broth alone or TYG broth containing either Tween 80 or citrate. Thus, the Tween 80 and citrate in the broth contributed to the commensalism. Metabolite analysis revealed that ethanol production in the co-metabolism of glucose and citrate by strain Y was suppressed by mixed culture in TYG broth containing Tween 80 and citrate, compared with that in TYG broth containing citrate alone. The mechanism supporting the observed commensal symbiosis between strains Y and IFO 13712 was the increase in availability of glucose for lactate production by strain Y because, in glycolysis, the pathway from glucose to lactate is energic, whereas the pathway from glucose to ethanol is not. Whether growth stimulation of strain Y by mixing it with IFO 13712 in milk products will enhance the survival of strain Y in the intestine remains to be elucidated.

Screening of Actinomycetes from Mangrove Ecosystem for L-asparaginase Activity and Optimization by Response Surface Methodology

Marine actinomycetes were isolated from sediment samples collected from Pitchavaram mangrove ecosystem situated along the southeast coast of India. Maximum actinomycete population was noted in rhizosphere region. About 38% of the isolates produced L-asparaginase. One potential strain KUA106 produced higher level of enzyme using tryptone glucose yeast extract medium. Based on the studied phenotypic characteristics, strain KUA106 was identified as Streptomyces parvulus KUA106. The optimization method that combines the Plackett-Burman design, a factorial design and the response surface method, which were used to optimize the medium for the production of L-asparaginase by Streptomycetes parvulus. Four medium factors were screened from eleven medium factors by Plackett-Burman design experiments and subsequent optimization process to find out the optimum values of the selected parameters using central composite design was performed. Asparagine, tryptone, d) extrose and NaCl components were found to be the best medium for the L-asparaginase production. The combined optimization method described here is the effective method for screening medium factors as well as determining their optimum level for the production of L-asparaginase by Streptomycetes parvulus KUAP106.

Caloramator mitchellensis sp. nov., a thermoanaerobe isolated from the geothermal waters of the Great Artesian Basin of Australia, and emended description of the genus Caloramator

A strictly thermophilic anaerobe, designated strain VF08(T), was isolated from a water sample collected from a Great Artesian Basin bore (registered bore number 22981) situated at Mitchell, QLD, Australia. Cells of isolate VF08(T) were slightly curved, non-sporulating rods (1.5-3.5 x 0.4-0.8 μm), which stained Gram-negative but possessed a Gram-positive cell-wall ultrastructure. The strain grew optimally in tryptone-yeast extract-glucose (TYEG) medium at 55 °C (temperature growth range between 37 and 60 °C) and a pH of 7 (pH growth range, 6.0-9.0). Yeast extract or tryptone was required for growth on glucose, fructose, xylose, maltose, sucrose, raffinose, cellobiose, ribose, pyruvate, tryptone, peptone, Casamino acids, amyl media and serine, but could also support growth as the sole carbon source. End products from glucose fermentation were acetate, ethanol, CO₂ and H₂. The strain reduced vanadium(V), but not iron(III), manganese(IV), elemental sulfur, sulfate, thiosulfate, sulfite, nitrate or nitrite in the presence of 0.2 % yeast extract, peptone, tryptone, glucose, sucrose and Casamino acids, but an increase in the growth rate or cell yield was not observed. Growth was inhibited by chloramphenicol, streptomycin, tetracycline, penicillin, ampicillin and ≥ 2 % NaCl (w/v). The G+C content of the DNA was 38.4 ± 0.8 mol% as determined by the thermal denaturation (T(m)) method. 16S rRNA gene sequence analysis revealed that isolate VF08(T) was a member of the genus Caloramator with Caloramator australicus and Caloramator fervidus (formerly Clostridium fervidus) being the closest relatives with similarity values of 85.0 and 86.1 %, respectively, when helix 6 nucleotides were included in the analysis, and 95.2 % and 94 %, respectively, when these nucleotides were masked from the analysis. Further analysis revealed that strain VF08(T) formed an individual cluster (cluster II) within the genus Caloramator and could be distinguished from other species within the genus Caloramator (clusters I, III and IV) on the basis of signature nucleotides and differences in phenotypic traits. These data suggest that strain VF08(T) is a novel species of the genus Caloramator, for which the name Caloramator mitchellensis sp. nov. is proposed. The type strain is VF08(T) (=JCM 15828(T)=KCTC 5735(T)). An emended description of the genus Caloramator is also provided.

Production of Extracellular Anti-leukaemic Enzyme Lasparaginase from Marine Actinomycetes by Solidstate and Submerged Fermentation: Purification and Characterisation

Purpose: The objective of this investigation was to isolate marine actinomycetes, screen them for Lasparaginase activity and characterise the enzyme.Methods: Marine actinomycetes were isolated from sediment samples obtained from Tamilnadu and Kerala in India. The isolates were identified as actinomycetes by microscopical and biochemical tests.Production of L-asparaginase was carried out in three different media, namely, solid-state media, Tryptone Glucose Yeast extract (TGY) broth and Tryptone Fructose Yeast extract (TFY) broth.. The enzyme was purified to near homogeneity by ammonium sulphate precipitation, dialysis, gel filtration on Sephadex G-100 column and SDS-PAGE.Results: Among 10 marine isolates subjected to preliminary screening, only isolates S3, S4 and K8 showed potential for L-asparaginase activity. All three marine soil isolates synthesized asparaginase with yield ranging from 24.6 to 49.2 IU/ml. Soil isolate S3 showed the highest productivity of 49.2 IU/ml with a protein content of 65 μg/ml and optimum activity at pH 7.5 and 50 ºC. The apparent Km value for the substrate was 25 μM. Mg2+ ion slightly stimulated activity while Cu2+, Zn2+ and EDTA were inhibitory.Conclusion: The study revealed that marine actinomycetes may be a potential source of high yield, high substrate specificity L-asparaginase, which is an anti-leukaemia agent.

Improving Solid-State Fermentation of Monascus purpureus on Agricultural Products for Pigment Production

In the present study, the effects of culture medium and temperature on red pigment production and mycelia growth were evaluated. The maximum red pigment production was found when Monascus purpureus CMU001 was cultivated on potato dextrose broth at 30 °C for 2 weeks. The highest amount of dry weight was achieved when cultivated on tryptone glucose yeast extract medium. Cheap agricultural products and residues were used as substrates for pigment production. Corn meal was the best substrate for pigment production (19.4 U/gds) when compared to peanut meal, coconut residue, and soybean meal. The highest pigment yield (129.63 U/gds) was found when corn meal was supplemented with 8% (w/w) glucose, followed by coconut residue (63.50 U/gds), peanut meal (52.50 U/gds), and soybean meal (22.50 U/gds). Galactose, sorbose, psicose, and mannitol were found to be good supplements next to glucose but not xylitol. Pigment was not stable at high temperature and long exposure to UV. The intensity of red pigment decayed 30.57% and 5.41% after autoclaving and pasteurization, respectively.

Deinococcus mumbaiensissp. nov., a radiation-resistant pleomorphic bacterium isolated from Mumbai, India

A radiation-resistant, Gram-negative and pleomorphic bacterium (CON-1) was isolated from a contaminated tryptone glucose yeast extract agar plate in the laboratory. It was red pigmented, nonmotile, nonsporulating, and aerobic, and contained MK-8 as respiratory quinone. The cell wall of this bacterium contained ornithine. The major fatty acids were C16:0, C16:1, C17:0, C18:1 and iso C18:0. The DNA of CON-1 had a G+C content of 70 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that CON-1 exhibited a maximum similarity (94.72%) with Deinococcus grandis. Based on the genotypic, phenotypic and chemotaxonomic characteristics, the bacterium CON-1 was identified as a new species of the genus Deinococcus, for which the name Deinococcus mumbaiensis sp. nov. is proposed. The type strain of D. mumbaiensis is CON-1 (MTCC 7297(T)=DSM 17424(T)).

Prevalence and characterization of Clostridium perfringens from spices in Argentina

Spices can present high microbial counts and Clostridium perfringens, Bacillus cereus, Salmonella and Shigella, among others have been isolated from spices. C. perfringens is an important pathogen agent causing, among other diseases, enteritis in humans caused by C. perfringens enterotoxin (CPE) which causes human food poisoning and enterotoxemia in domestic animals. The aims of the present work were (i) to establish the hygienic sanitary quality of some spices in San Luis, Argentina; (ii) to determine the presence of C. perfringens in these spices by means of the most probable number (MPN) and count on plate methods; (iii) to characterize the enterotoxigenic strains of C. perfringens by PCR and immunological methods such as reverse passive latex agglutination (RPLA) and (iv) to type by PCR C. perfringens strains isolated. A total of 115 samples of spices, 67 of which were purchased in local retail stores and 48 domestically collected were analysed. Total aerobe counts on tryptone glucose yeast extract agar medium of the 115 samples were between <10 and 10 6 CFU/g. The colifecal counts using Mac Conkey broth of the 115 samples were <4–10 3 CFU/g, with 28 samples (24.34%) exceeding the limit established by the Spanish Alimentary Code (10 CFU/g) while 2 samples (1.73%) had a sulfite reducing anaerobe load above standard limits. A total of 14 C. perfringens strains (12.17%) were isolated and characterized from 115 samples by the standard biochemical tests. Four of which (28.60%) turned out to be enterotoxigenic by PCR and RPLA. In order to type C. perfringens strains based on their main toxins, the 14 strains were analysed by PCR. All strains belonged to type A. All RPLA positive strains were cpe + by PCR. The percentage of enterotoxigenic strains was more elevated that those reported in other studies for this type of sample. These results indicate that sanitary conditions in different production stages of species must be improved to reduce health hazards. The high percentage (24.34%) of samples with colifecal values above standard limits is an indication of deficient sanitary conditions. These results suggest the need to provide legislation on the sanitary and hygienic quality of spices in our country.

PH effects on 10 Streptomyces spp. growth and sporulation depend on nutrients

Streptomycetes are regarded to prefer neutral to alkaline environmental pH, although they commonly occur at remarkably variable pH and nutritional conditions. Therefore, the dependence of 10 Streptomyces spp. pH tolerance on nutrients was determined. Ten environmental Streptomyces spp. were grown and sporulated between pH 4.0 and 11.5, at the interval of 1.5, on starch-casein-KNO(3), tryptone-yeast extract-glucose, glycerol-arginine and tryptone-soy agars, and three their modifications. On media with starch and casein; glucose, tryptone and yeast extract; tryptone and soy peptone; and glycerol-arginine and yeast extract strains grew over a broad pH range between 4.0-5.5 and 10.0-11.5. On glycerol-arginine and on medium with Na-propionate, NH(4)NO(3) and yeast extract, streptomycetes grew optimally at pH 7.0 and above. The high organic load enabled the growth over a wide pH range. The sporulation pH ranges followed those for growth. The high organic load enabled the growth over a wide pH range. The strain-specific differences in sporulation were greater than those caused by pH. The best medium for sporulation contained glucose and tryptone with minerals of glycerol-arginine agar at pH 5.5. The growth pH ranges, pH ranges for the optimal growth, and sporulation were strongly dependent on nutrients.

Performance of mycological media in enumerating desiccated food spoilage yeasts: an interlaboratory study

Dichloran 18% glycerol agar (DG18) was originally formulated to enumerate nonfastidious xerophilic moulds in foods containing rapidly growing Eurotium species. Some laboratories are now using DG18 as a general purpose medium for enumerating yeasts and moulds, although its performance in recovering yeasts from dry foods has not been evaluated. An interlaboratory study compared DG18 with dichloran rose bengal chloramphenicol agar (DRBC), plate count agar supplemented with chloramphenicol (PCAC), tryptone glucose yeast extract chloramphenicol agar (TGYC), acidified potato dextrose agar (APDA), and orange serum agar (OSA) for their suitability to enumerate 14 species of lyophilized yeasts. The coefficient of variation for among-laboratories repeatability within yeast was 1.39% and reproducibility of counts among laboratories was 7.1%. The order of performance of media for recovering yeasts was TGYC>PCAC=OSA>APDA>DRBC>DG18. A second study was done to determine the combined effects of storage time and temperature on viability of yeasts and suitability of media for recovery. Higher viability was retained at −18°C than at 5°C or 25°C for up to 42 weeks, although the difference in mean counts of yeasts stored at −18°C and 25°C was only 0.78 log 10 cfu/ml of rehydrated suspension. TGYC was equal to PCAC and superior to the other four media in recovering yeasts stored at −18°C, 5°C, or 25°C for up to 42 weeks. Results from both the interlaboratory study and the storage study support the use of TGYC for enumerating desiccated yeasts. DG18 is not recommended as a general purpose medium for recovering yeasts from a desiccated condition.

Comparison of dichloran 18% glycerol (DG18) agar with general purpose mycological media for enumerating food spoilage yeasts

Dichloran 18% glycerol (DG18) agar was originally developed to enumerate xerophilic foodborne moulds. However, some laboratories are using DG18 agar as a general medium to enumerate foodborne moulds and yeasts. A collaborative study, with the participation of seven laboratories, was undertaken to compare DG18 agar with dichloran rose bengal chloramphenicol (DRBC) agar, tryptone glucose yeast extract chloramphenicol (TGYC) agar, and plate count agar supplemented with chloramphenicol (PCAC) for enumerating 14 species of common food spoilage yeasts. Comparison of the mean values of populations of all yeasts recovered on each medium revealed no significant differences among DRBC agar, PCAC, and TGYC agar, while each of these media supported the development of significantly ( P≤0.05) higher numbers of colonies than DG18 agar. However, differences were only 0.08 to 0.10 log 10 cfu/ml, making the practical significance questionable. The overall coefficient of variation (CV) for within laboratory repeatability was 1.71%, while the CV for reproducibility of counts obtained among laboratories was 6.96%. Compared to DRBC agar, TGYC agar, and PCAC, yeast colonies were smaller on DG18 agar. Growth of Brettanomyces anomalus, Cryptococcus albidus, and Rhodotorula mucilaginosa was particularly retarded or inhibited on DG18 agar. Based on the performance of media in supporting colony development and ease of counting colonies, the use of DG18 agar as a general enumeration medium for foodborne yeasts cannot be recommended.

Presence and changes in populations of yeasts on raw and processed poultry products stored at refrigeration temperature

A study was undertaken to determine populations and profiles of yeast species on fresh and processed poultry products upon purchase from retail supermarkets and after storage at 5°C until shelf life expiration, and to assess the potential role of these yeasts in product spoilage. Fifty samples representing 15 commercial raw, marinated, smoked, or roasted chicken and turkey products were analyzed. Yeast populations were determined by plating on dichloran rose bengal chloramphenicol (DRBC) agar and tryptone glucose yeast extract (TGY) agar. Proteolytic activity was determined using caseinate and gelatin agars and lipolytic activity was determined on plate count agar supplemented with tributyrin. Populations of aerobic microorganisms were also determined. Initial populations of yeasts (log 10 cfu/g) ranged from less than 1 (detection limit) to 2.89, and increased by the expiration date to 0.37–5.06, indicating the presence of psychrotrophic species. Highest initial populations were detected in raw chicken breast, wings, and ground chicken, as well as in turkey necks and legs, whereas roasted chicken and turkey products contained less than 1 log 10 cfu/g. During storage, yeast populations increased significantly ( P≤0.05) in whole chicken, ground chicken, liver, heart and gizzard, and in ground turkey and turkey sausage. Isolates (152 strains) of yeasts from poultry products consisted of 12 species. Yarrowia lipolytica and Candida zeylanoides were predominant, making up 39 and 26% of the isolates, respectively. Six different species of basidiomycetous yeasts representing 24% of the isolates were identified. Most Y. lipolytica strains showed strong proteolytic and lipolytic activities, whereas C. zeylanoides was weakly lipolytic. Results suggest that yeasts, particularly Y. lipolytica, may play a more prominent role than previously recognized in the spoilage of fresh and processed poultry stored at 5°C.

Changes in acid tolerance of Lactococcus lactis during growth at constant pH

Cells of Lactococcus lactis MG1363 growing in batch culture in TYG (tryptone, yeast extract, glucose) medium at constant pH 7.0 became gradually more acid sensitive shortly after inoculation until a point of maximum sensitivity was reached in early log-phase. The acid tolerance then gradually increased in the mid- and late-log phase until maximum tolerance was reached at the onset of stationary phase. This pattern has been termed the growth-phase acid tolerance. The variation in acid tolerance seen in pH 7.0 grown cells of L. lactis MG1363 did not result from changes in internal pH or membrane H +ATPase activity levels. Neither the amount of glucose present during mid-log phase nor the amount of lactate produced by the cells correlated with the pattern of the log-phase acid tolerance. Cells grown in partially spent TYG medium showed a reduced growth rate and increased acid tolerance compared to cells grown in fresh TYG medium. Supplementing the spent medium with tryptone or yeast extract or both restored the growth rate and cells became more sensitive to acid. Fractionation of tryptone yielded a fraction which stimulated the growth of MG1363 in partially spent medium and delayed the acquisition of acid tolerance. The active compound(s) has a putative molecular weight of about 1 kDa and was partially degraded by papain and trypsin.

A collaborative study on media for the enumeration of yeasts in foods

A collaborative study was made to evaluate the effectivity of a general purpose medium, tryptone glucose yeast extract (TGY) agar on the detection and enumeration of yeasts from food. Nine laboratories participated in the study and compared five media (four kinds of TGY with different concentrations of glucose, one of them without tryptone, and, for comparison, dichloran rose bengal chloramphenicol (DRBC) agar). Six food samples were investigated by each laboratory and 23 additional food samples were investigated individually by different laboratories. No difference was found in the performance of media with either the samples common to all laboratories or the various samples tested in different ones. A medium consisting of tryptone, glucose and yeast extract, at any concentration of glucose tested, appeared reliable for the detection and enumeration of yeasts from foods, and its performance did not differ from that of DRBC. Omission of tryptone as recommended by ISO provided an even simpler medium of equally good performance. TGY without chloramphenicol may result in higher total counts due to the development of bacteria. DRBC incubated in light results in lower counts compared to that incubated in the dark.

Influence of Growth Medium on Thermal Resistance of Pediococcus sp. NRRL B-2354 (Formerly Micrococcus freudenreichii) in Liquid Foods

Pediococcus sp. is a nonpathogenic heat-resistant spoilage organism that has been used as a test organism in milk pasteurization studies. These characteristics make this bacterium an attractive test organism to study the mode of bacterial thermal inactivation in a food pilot plant. We report here the effect of growth medium on the thermal D value of this organism in skim milk, whole liquid egg, 10% glucose solution, pineapple juice, apple juice, tomato juice, and water at 60°C. Thermal inactivation was done in a submerged coil; D values were calculated from the linear portion of the survival curves by linear regression analysis. The range of D values of stationary-phase cells grown at 28°C in tryptone glucose yeast extract (TGY) or tryptic soy broth (TSB) was 0.14 to 12.05 min in all heating menstrua tested. The TSB-grown cells exhibited the highest thermal resistance with skim milk and 10% glucose solution as the heating menstrua. Survival curves of the TGY-grown cells indicated the presence of a cell population heterogeneous in thermal resistance. The TSB-grown cells exhibited a cell population uniform in thermal resistance and with a lag time for thermal inactivation. When compared to TGY-grown cells, Pediococcus sp. grown in TSB showed a significant (P < 0.05) increase in D values by up to eightfold in all heating menstrua. Results from this study suggested that thermal inactivation of Pediococcus sp. was dependent on the growth medium and on the heating menstruum with respect to both pH and composition.

Evaluation of media for enumerating heat-stressed, benzoate-resistant Zygosaccharomyces bailii

A study was undertaken to determine the efficacy of yeast malt extract agar (YMA), acidified YMA (YMAA), acidified tryptone glucose yeast extract agar (TGYA) and Zygosaccharomyces bailii agar (ZBA) for enumerating two strains of Z. bailii (FRR 2227 and FRR 3680) grown and heated in blueberry syrup containing 0, 300 and 600 μg ml sodium benzoate. Cells were heated at 50, 51 or 52 °C for 0, 15, 30 or 45 min before spread plating diluted samples on enumeration media and incubating at 30 °C for 5 days. Strain FRR 3680 exhibited higher heat resistance than did strain FRR 2227. Regardless of the sodium benzoate concentration in growth and heating media, or the temperature and time of heating, non-selective YMA supported recovery of the highest number of viable cells, followed by TGYA for strain FRR 2227 and TGYA or ZBA for strain FRR 3680. YMAA supported the lowest recovery of heat-stressed cells. The concentration of sodium benzoate in the growth medium did not have a significant effect on tolerance of strain FRR 3680 to heat; however, tolerance of strain FRR 2227 to heat decreased when cells were grown in blueberry syrup containing increased concentrations of sodium benzoate. Regardless of the sodium benzoate concentration in the growth medium, both strains exhibited increased sensitivity when heated in blueberry syrup containing increased concentrations of sodium benzoate. Overall, TGYA was judged best among selective media for enumerating Z. bailii because of its ability to support colony development by heat-stressed cells and because of the ease of counting and differentiating colonies.