Characterization and probiotic potential of Lactobacillus plantarum strains isolated from cheeses

Ninety-eight Lactobacillus plantarum strains isolated from Italian and Argentinean cheeses were evaluated for probiotic potential. After a preliminary subtractive screening based on the presence of msa and bsh genes, 27 strains were characterized. In general, the selected strains showed high resistance to lysozyme, good adaptation to simulated gastric juice, and a moderate to low bile tolerance. The capacity to agglutinate yeast cells in a mannose-specific manner, as well as the cell surface hydrophobicity was found to be variable among strains. Very high β-galactosidase activity was shown by a considerable number of the tested strains, whereas variable prebiotic utilization ability was observed. Only tetracycline resistance was observed in two highly resistant strains which harbored the tetM gene, whereas none of the strains showed β-glucuronidase activity or was capable of inhibiting pathogens. Three strains (Lp790, Lp813, and Lp998) were tested by in vivo trials. A considerable heterogeneity was found among a number of L. plantarum strains screened in this study, leading to the design of multiple cultures to cooperatively link strains showing the widest range of useful traits. Among the selected strains, Lp790, Lp813, and Lp998 showed the best probiotic potential and would be promising candidates for inclusion as starter cultures for the manufacture of probiotic fermented foods.     

Investigation of the factors influencing the survival of Bifidobacterium longum in model acidic solutions and fruit juices

The survival of Bifidobacterium longum NCIMB 8809 was studied during refrigerated storage for 6 weeks in model solutions, based on which a mathematical model was constructed describing cell survival as a function of pH, citric acid, protein and dietary fibre. A Central Composite Design (CCD) was developed studying the influence of four factors at three levels, i.e., pH (3.2–4), citric acid (2–15 g/l), protein (0–10 g/l), and dietary fibre (0–8 g/l). In total, 31 experimental runs were carried out. Analysis of variance (ANOVA) of the regression model demonstrated that the model fitted well the data. From the regression coefficients it was deduced that all four factors had a statistically significant (P < 0.05) negative effect on the log decrease [log₁₀N_{0 week}−log₁₀N_{6 week}], with the pH and citric acid being the most influential ones. Cell survival during storage was also investigated in various types of juices, including orange, grapefruit, blackcurrant, pineapple, pomegranate and strawberry. The highest cell survival (less than 0.4 log decrease) after 6 weeks of storage was observed in orange and pineapple, both of which had a pH of about 3.8. Although the pH of grapefruit and blackcurrant was similar (pH ∼3.2), the log decrease of the former was ∼0.5 log, whereas of the latter was ∼0.7 log. One reason for this could be the fact that grapefruit contained a high amount of citric acid (15.3 g/l). The log decrease in pomegranate and strawberry juices was extremely high (∼8 logs). The mathematical model was able to predict adequately the cell survival in orange, grapefruit, blackcurrant, and pineapple juices. However, the model failed to predict the cell survival in pomegranate and strawberry, most likely due to the very high levels of phenolic compounds in these two juices.     

Diversity of stress tolerance in Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus paraplantarum: A multivariate screening study

Sixty-three strains of the taxonomically related species Lactobacillus plantarum subsp. plantarum, L. plantarum subsp. argentoratensis, L. paraplantarum and L. pentosus isolated from sourdoughs and other food and non-food sources and 14 strains of other members of the genus Lactobacillus were screened for their tolerance of acid, alkaline, heat, oxidative, osmotic, detergent and starvation stresses in order to evaluate the diversity of stress response. Most strains of the L. plantarum group were highly tolerant of acid, alkaline and osmotic stress and highly sensitive to detergent stress, while a larger diversity was found for other stress. Multivariate analysis allowed grouping the strains in clusters with similar response patterns. Stress response patterns in the L. plantarum group were similar to those of species of the L. casei/L. paracasei group but clearly different from those of other mesophilic Lactobacillus. No relationship was found between grouping obtained on the basis of stress response patterns and by genotypic fingerprinting (rep-PCR), nor with the taxonomic position or isolation source of the strains. Further experiments with selected strains showed that exponential phase cells were generally but not always more sensitive than stationary phase cells. The ability to grow under stressful conditions showed a slightly better correlation with the ecological conditions prevailing in the isolation niches of the strains.This study will be the basis for further investigations to identify and exploit the basis of diversity in the stress response of lactic acid bacteria.     

The effect of Lactobacillus buchneri and Lactobacillus plantarum on the fermentation, aerobic stability, and ruminal degradability of low dry matter corn and sorghum silages

The effect of Lactobacillus buchneri, alone or in combination with Lactobacillus plantarum, on the fermentation, aerobic stability, and ruminal degradability of low dry matter corn and sorghum silages was studied under laboratory conditions. The inoculants were applied at 1 x 10(6) cfu/g. Silages with no additives served as control. After treatment, the chopped forages were ensiled in 1.5-L anaerobic jars. Three jars per treatment were sampled on d 2, 4, 8, 15, and 90. After 90 d of storage, the silages were subjected to an aerobic stability test lasting 5 d, in which CO2 production, as well as chemical and microbiological parameters, was measured to determine the extent of aerobic deterioration. At the end of the ensiling period (d 90), the L. buchneri- and L. buchneri + L. plantarum-inoculated silages had significantly higher levels of acetic acid than the control and L. plantarum-inoculated silages. Therefore, yeast activity was impaired in the L. buchneri- and L. buchneri + L. plantarum-inoculated silages. As a result, L. buchneri, alone or in combination with L. plantarum, improved aerobic stability of the low dry matter corn and sorghum silages. The combination of L. buchneri and L. plantarum reduced ammonia N concentrations and fermentation losses in the silages compared with L. buchneri alone. However, L. buchneri, L. plantarum, and a combination of L. buchneri + L. plantarum did not effect in situ rumen dry matter, organic matters, or neutral detergent fiber degradability of the silages. The L. buchneri was very effective in protecting the low dry matter corn and sorghum silages exposed to air under laboratory conditions. The use of L. buchneri, alone or in combination with L. plantarum, as a silage inoculant can improve the aerobic stability of low dry matter corn and sorghum silages by inhibition of yeast activity.     

Binding of mutagens to exopolysaccharide produced by Lactobacillus plantarum mutant strain 301102S

Exopolysaccharide (EPS) was produced by Lactobacillus plantarum 301102 on exposure to the mutagenic action of acridine orange and novobiocin. The biological characteristics of this mutant strain 301102S were the same as those of the parent strain, but fermented milk prepared with the mutant strain showed antimutagenic activity on 3-amino-1,4-dimethyl-5H-pyrido indole. Only EPS-bound cells of strain 301102S showed binding ability to mutagens such as heterocyclic amines, and the mutagens were inactivated by binding to EPS. The binding ability was affected by pH; the greatest percentage binding was noted at pH 8.0. Addition of Mg²⁺ and sodium dodecyl sulfate, but not oxgall, inhibited the binding ability. Therefore, the binding mechanism of the EPS may consist of ion-exchange and hydrophobic bonds, and the EPS would bind mutagens in the intestine.     

Coculture-inducible bacteriocin activity of Lactobacillus plantarum strain J23 isolated from grape must

Detection and characterization of bacteriocin production by Lactobacillus plantarum strain J23, recovered from a grape must sample in Spain, have been carried out. Bacteriocin activity was degraded by proteolytic enzymes (trypsin, alfa-chymotrypsin, papaine, protease, proteinase K and acid proteases), and it was stable at high temperatures (121 degrees C, 20min), in a wide range of pH (1-12), and after treatment with organic solvents. L. plantarum J23 showed antimicrobial activity against Oenococcus oeni, and a range of Lactobacillus and Pediococcus species. Bacteriocin production was detected in liquid media only when J23 was cocultivated with some inducing bacteria, and induction took place when intact cells or 55 degrees C heated cells of the inducer were cocultivated with J23, but not with their autoclaved cells. Bacteriocin activity of J23 was not induced by high initial J23 inocula, and it was detected in cocultures during the exponential phase. The presence of ethanol or acidic pH in the media reduced bacteriocin production in the cocultures of J23 with the inducing bacteria. The presence of plantaricin-related plnEF and plnJ genes was detected by PCR and sequencing. Nevertheless, negative results were obtained for plnA, plnK, plNC8, plS and plW genes.     

Production of salami from beef, horse, mutton, Blesbok (Damaliscus dorcas phillipsi) and Springbok (Antidorcas marsupialis) with bacteriocinogenic strains of Lactobacillus plantarum and Lactobacillus curvatus

Lactobacillus plantarum 423, producer of bacteriocin 423, Lactobacillus curvatus DF38, producer of curvacin DF38, and a bacteriocin-negative mutant of L. plantarum 423 (423m) were evaluated as starter cultures in the production of salami from beef, horse, mutton, Blesbok (Damaliscus dorcas phillipsi) and Springbok (Antidorcas marsupialis). Growth of L. plantarum 423 and L. curvatus DF38 was best supported in Blesbok salami, as revealed by the highest growth rate during sweating, cold smoking and maturation, and final cell numbers after 70 days (1 × 10⁸ and 5 × 10⁷ cfu/g, respectively). Growth of Listeria innocua was the best suppressed in Blesbok salami fermented with L. plantarum 423 and L. curvatus DF38. Growth of L. innocua in horse salami was best suppressed when fermented with L. curvatus DF38. The final pH of salami fermented with L. plantarum 423 and L. plantarum 423m was slightly lower (4.4) compared to the pH of salami produced with L. curvatus DF38 (pH 4.7). No significant differences (P > 0.05) were recorded by a trained sensory taste panel amongst the three starter cultures regarding colour and venison like aroma. Horse, Blesbok and Springbok salami were rated significantly higher (P ? 0.05) in salami flavour than mutton salami, which was rated the lowest for this attribute. Blesbok salami was rated the highest for sour meat aroma, while beef salami was rated the lowest. Springbok salami was rated the highest in terms of oily mouth feel. Beef salami had the most compact structure and horse salami the softest structure of all meat types fermented. In general, salami produced with L. plantarum 423 yielded the best sour meat aroma, colour, texture, venison like flavour, sour meat flavour and oily mouthfeel and is considered superior to the L. plantarum mutant (strain 423m) and L. curvatus DF38.     

Antioxidant activity of Lactobacillus plantarum strains isolated from traditional Chinese fermented foods

Eleven Lactobacillus plantarum strains isolated from traditional Chinese fermented foods were investigated for their in vitro scavenging activity against hydroxyl and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals, and their resistance to hydrogen peroxide. L. plantarum C88 at a dose of 10(10) CFU/ml showed the highest hydroxyl radical and DPPH scavenging activities, with inhibition rates of 44.31% and 53.05%, respectively. Resistance of intact cells to hydrogen peroxide was also found in all strains. L. plantarum C88 was the most resistant strain against hydrogen peroxide. When L. plantarum C88 was administered to senescent mice suffering oxidative stress induced by d-galactose, the serum superoxide dismutase activity, the glutathione peroxidase activity and the total antioxidant capacity in liver increased significantly, while the level of malondialdehyde in liver decreased significantly. L. plantarum C88 isolated from traditional Chinese fermented dairy tofu could be considered as a potential antioxidant to be applied in functional foods.     

Mixed species biofilms of Listeria monocytogenes and Lactobacillus plantarum show enhanced resistance to benzalkonium chloride and peracetic acid

We investigated the formation of single and mixed species biofilms of Listeria monocytogenes strains EGD-e and LR-991, with Lactobacillus plantarum WCFS1 as secondary species, and their resistance to the disinfectants benzalkonium chloride and peracetic acid. Modulation of growth, biofilm formation, and biofilm composition was achieved by addition of manganese sulfate and/or glucose to the BHI medium. Composition analyses of the mixed species biofilms using plate counts and fluorescence microscopy with dual fluorophores showed that mixed species biofilms were formed in BHI (total count, 8-9 log₁₀ cfu/well) and that they contained 1-2 log₁₀ cfu/well more L. monocytogenes than L. plantarum cells. Addition of manganese sulfate resulted in equal numbers of both species (total count, 8 log₁₀ cfu/well) in the mixed species biofilm, while manganese sulfate in combination with glucose, resulted in 1-2 log₁₀ more L. plantarum than L. monocytogenes cells (total count, 9 log₁₀ cfu/well). Corresponding single species biofilms of L. monocytogenes and L. plantarum contained up to 9 log₁₀ cfu/well. Subsequent disinfection treatments showed mixed species biofilms to be more resistant to treatments with the selected disinfectants. In BHI with additional manganese sulfate, both L. monocytogenes strains and L. plantarum grown in the mixed species biofilm showed less than 2 log₁₀ cfu/well inactivation after exposure for 15 min to 100 μg/ml benzalkonium chloride, while single species biofilms of both L. monocytogenes strains showed 4.5 log₁₀ cfu/well inactivation and single species biofilms of L. plantarum showed 3.3 log₁₀ cfu/well inactivation. Our results indicate that L. monocytogenes and L. plantarum mixed species biofilms can be more resistant to disinfection treatments than single species biofilms.     

Survival of human derived Lactobacillus plantarum in fermented cereal extracts during refrigerated storage

The aim of this study was to investigate the survival of a potentially probiotic Lactobacillus plantarum strain in barley, wheat and barley malt extracts. The extracts were produced from three flour/water suspensions, i.e., 5%, 20%, 30% w/w. After inoculation, the cultures were incubated for 24 h at 37 °C, and were subsequently stored at 4 °C for up to seventy days. The lactic acid and reducing sugar concentrations at the beginning of storage were significantly different between the fermented media, ranging from 0.5 g/L to 17 g/L and from 0.8 g/L to 6.5 g/L respectively, while the pH ranged between 2.9 and 3.4. It was observed that the cells survived much better in the malt extracts compared to barley and wheat extracts during refrigerated storage. Based on the results from a study using model media and supplemented cereal extracts it was derived that this was most likely due to their higher sugar concentration and the presence of protective unidentified compounds, albeit the fact that the malt extracts contained higher amounts of lactic acid.     

The growth of Propionibacterium cyclohexanicum in fruit juices and its survival following elevated temperature treatments

This study investigated the growth of Propionibacterium cyclohexanicum in orange juice over a temperature range from 4 to 40 degrees C and its ability to multiply in tomato, grapefruit, apple, pineapple and cranberry juices at 30 and 35 degrees C. Survival after 10 min exposure to 50, 60, 70, 80, 85, 90 and 95 degrees C in culture medium and in orange juice was also assessed. In orange juice the organism was able to multiply by 2 logs at temperatures from 4 to 35 degrees C and survived for up to 52 days. However, at 40 degrees C viable counts were reduced after 6 days and no viable cells isolated after 17 days. The optimum growth temperature in orange juice over 6 days was 25 degrees C but over 4 days it was 35 degrees C. The growth of P. cyclohexanicum was monitored in tomato, grapefruit, cranberry, pineapple and apple juices at 30 and 35 degrees C over 29 days. Cranberry, grapefruit and apple juice did not support the growth of P. cyclohexanicum. At 30 degrees C no viable cells were detected after 8 days in cranberry juice or after 22 days in grapefruit juice while at 35 degrees C no viable cells were detected after 5 and 15 days, respectively. However, in apple juice, although a 5 log reduction occurred, viable cells could be detected after 29 days. P. cyclohexanicum was able to multiply in both tomato and pineapple juices. In tomato juice, there was a 2 log increase in viable counts after 8 days at 30 degrees C but no increase at 35 degrees C, while in pineapple juice there was a 1 log increase in numbers over 29 days with no significant difference between numbers of viable cells present at 30 and 35 degrees C. The organism survived at 50 degrees C for 10 min in culture medium without a significant loss of viability while similar treatment at 60, 70 and 80 degrees C resulted in approximately a 3-4 log reduction, with no viable cells detected after treatment at 85 or 90 or 95 degrees C but, when pre-treated at intermediate temperatures before exposure to higher temperatures, some cells survived. However, in orange juice a proportion of cells survived at 95 degrees C for 10 min without pre-treatment and there was no significant difference between numbers surviving with and without pre-treatment. The results from this study demonstrate that P. cyclohexanicum is able to grow in a number of juices, other than orange juice, and able to survive a number of high temperature procedures. Therefore, if initially present in the raw materials P. cyclohexanicum might survive the pasteurization procedures used in the fruit juice industry, contaminate and consequently spoil the final product.     

Bacterial inactivation in fruit juices using a continuous flow Pulsed Light (PL) system

In this work, the susceptibility to pulsed light (PL) treatments of both a Gram-positive (L. innocua 11288) and a Gram-negative (E. coli DH5-??) bacteria inoculated in apple (pH = 3.49, absorption coefficient 13.9 cm^− 1) and orange juices (pH = 3.78, absorption coefficient 52.4 cm^− 1) was investigated in a range of energy dosages from 1.8 to 5.5 J/cm². A laboratory scale continuous flow PL system was set up for the experiments, using a xenon flash-lamp emitting high intensity light in the range of 100-1100 nm. The flashes lasted 360 ??s at a constant frequency of 3 Hz.The results highlighted how the lethal effect of pulsed light depended on the energy dose supplied, the absorption properties of liquid food as well as the bacterial strain examined. The higher the quantity of the energy delivered to the juice stream, the greater the inactivation level. However, the absorbance of the inoculated juice strongly influenced the dose deliver and, therefore, the efficiency of the PL treatment. Among the bacteria tested, E. coli cells showed a greater susceptibility to the PL treatment than L. innocua cells in both apple and orange juices. Following treatment at 4 J/cm², microbial reductions in apple and orange juices were, respectively, 4.00 and 2.90 Log-cycles for E. coli and 2.98 and 0.93 Log-cycles for L. innocua.Sublethally injured cells were also detected for both bacterial strains, thus confirming that membrane damage is an important event in bacterial inactivation by PL.     

Effect of microencapsulation on survival of Lactobacillus plantarum in simulated gastrointestinal conditions, refrigeration, and yogurt

In the present research the survival of free and microencapsulated cells of a new strain of Lactobacillus plantarum BL011 under stress conditions was tested in sodium alginate or pectin, coated with sodium alginate or chitosan. Results for the simulated gastrointestinal medium (SGT) showed no change in viability of cells in relation to the control. However, the simulated gastric medium (GM) drastically reduced the viability under the tested conditions, with no significant differences between free and immobilized cells. Under refrigerated storage viability of immobilized cells were greatly enhanced compared to the free microorganisms, and the treatments showing the lowest loss of viability were those of 4% (w/v) pectin, 3% (w/v) sodium alginate coated with chitosan and a mixture of 2% (w/v) sodium alginate and 2% (w/v) pectin, respectively. Loss of viability of immobilized L. plantarum in 3% alginate coated with chitosan in yogurt was of 0.55 log cycles during 38 days of storage. The results of this study suggest the efficiency of immobilization techniques to increase the survival of lactobacilli in yogurt under refrigerated storage.     

Technological and probiotic role of adjunct cultures of non-starter lactobacilli in soft cheeses

The influence of two cheese-isolated Lactobacillus strains on cheese composition, acceptability and probiotic capacity was assessed. Soft cheeses with and without the addition of Lactobacillus plantarum I91 or Lactobacillus paracasei I90 were prepared. Gross composition was assessed and secondary proteolysis was described by soluble fractions and free amino acids profiles. Acceptability was determined by a panel of 98 non-trained consumers. Cheeses harboring added Lactobacillus strains were also studied in vivo to evaluate their probiotic capacity. Gross composition of the cheeses was similar for control and treated (Lactobacillus-added) cheeses. Peptidolysis increased in cheeses with added lactobacilli, which was evidenced by a higher free amino acid content. Overall, the acceptability of the cheeses was good: 65%–80% of the consumers said that they “liked very much” or “liked” the cheeses. Cheeses with L. plantarum I91 showed the highest changes in composition and proteolysis and were the most accepted ones. On the contrary, composition of cheeses with L. paracasei I90 was similar to that of the controls, but these samples were less accepted than cheeses without lactobacilli. The oral administration of cheese containing L. plantarum I91 or L. paracasei I90 proved to be safe and able to enhance the number of IgA + cells in the small intestine lamina propria of mice. The use of selected strains of NSLAB exerted a technological and probiotic role: it contributed to the standardization of cheese quality and induced benefic health effects at the gut mucosa in vivo.     

Technological and safety properties of Lactobacillus plantarum strains isolated from a Tunisian traditional salted meat

A total of 17 strains of Lactobacillus plantarum, isolated from a Tunisian traditional salted meat and identified by biochemical and molecular methods, were characterized according to their technological properties including acidifying, antimicrobial and enzymatic activities as well as antibiotic resistance in order to select the most suitable for use as starter cultures for the production of fermented sausages. All the strains studied showed good acidifying activity and were able to reduce the pH to less than 4.3 in 72, 48 and 24h at 15, 25 and 37°C respectively. The majority of strains displayed antimicrobial activities against Salmonella arizonae, Staphylococcus aureus, Pseudomonas aeuroginosa and Escherichia coli, however characterization of the antimicrobial substances showed that none of the strains could produce bacteriocins. All the L. plantarum strains were able to hydrolyze casein, whereas none of them was found to possess lipolytic activity. The majority of strains of L. plantarum were resistant to tetracycline, erythromycin, rifampicin, ampicillin and penicillin G.     

Nonthermal inactivation and sublethal injury of Lactobacillus plantarum in apple cider by a pilot plant scale continuous supercritical carbon dioxide system

The objective of this study was to evaluate the efficacy of supercritical carbon dioxide (SCCO₂) for inactivating Lactobacillus plantarum in apple cider using a continuous system with a gas-liquid metal contactor. Pasteurized apple cider without preservatives was inoculated with L. plantarum and processed using a SCCO₂ system at a CO₂ concentration range of 0-12% (g CO₂/100 g product), outlet temperatures of 34, 38, and 42 °C, a system pressure of 7.6 MPa, and a flow rate of 1 L/min. Processing with SCCO₂ significantly (P < 0.05) enhanced inactivation of L. plantarum in apple cider, resulting in a 5 log reduction with 8% CO₂ at 42 °C. The response surface model indicated that both CO₂ concentration and temperature contributed to the microbial inactivation. The extent of sublethal injury in surviving cells in processed apple cider increased as CO₂ concentration and processing temperature increased, however the percent injury dramatically decreased during SCCO₂ processing at 42 °C. Structural damage in cell membranes after SCCO₂ processing was observed by SEM. Refrigeration (4 °C) after SCCO₂ processing effectively inhibited the re-growth of surviving L. plantarum during storage for 28 days. Thus this study suggests that SCCO₂ processing is effective in eliminating L. plantarum and could be applicable for nonthermal pasteurization of apple cider.     

Alicyclobacillus acidoterrestris in pasteurized exotic Brazilian fruit juices: Isolation,genotypic characterization and heat resistance

In this study, the population of Alicyclobacillus spp. was estimated in pasteurized exotic Brazilian fruit juices using the most probable number (MPN) technique followed by biochemical tests. Pasteurized passion fruit (n = 57) and pineapple (n = 50) juices were taken directly from Brazilian manufacturers. While Alicyclobacillus spp. was isolated from passion fruit juice, the microorganism was not found in any pineapple juice samples. A higher incidence of Alicyclobacillus was observed in samples taken in June and July (dry months in Brazil) in comparison to the other months (March, April, May and August), and the highest Alicyclobacillus counts were recovered from these samples(>23 MNP/100 mL). Sixteen (n = 16) Alicyclobacillus strains were typed using the randomly amplified polymorphic DNA method (RAPD-PCR). RAPD-PCR revealed great genetic similarity between the passion fruit juice strains and Alicyclobacillus acidoterrestris DSM 2498. The heat resistance of three isolates was determined, and the mean D_95°  (1.7 min) and z (7.6 °C) values in the passion fruit juice were not significantly different (p > 0.05) from those obtained for the DSM 2498 strain (D_95°  = 1.5 min and z = 7.1 °C). This is the first report on the isolation of A. acidoterrestris from exotic fruit juices such as passion fruit juice. It is worth pointing out the importance of applying good agricultural practices in the field and applying controls for the fruit selection and washing steps, as well as controlling the time/temperature conditions for pasteurization so as to reduce the incidence and chances of A. acidoterrestris spoilage in these juices.     

Selective enumeration of Lactobacillus delbrueckii ssp. bulgaricus,Streptococcus thermophilus,Lactobacillus acidophilus,bifidobacteria, Lactobacillus casei,Lactobacillus rhamnosus,and propionibacteria

Nineteen bacteriological media were evaluated to assess their suitability to selectively enumerate Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, bifidobacteria, and propionibacteria. Bacteriological media evaluated included Streptococcus thermophilus agar, pH modified MRS agar, MRS-vancomycine agar, MRS-bile agar, MRS-NaCl agar, MRS-lithium chloride agar, MRS-NNLP (nalidixic acid, neomycin sulfate, lithium chloride and paramomycine sulfate) agar, reinforced clostridial agar, sugar-based (such as maltose, galactose, sorbitol, manitol, esculin) media, sodium lactate agar, arabinose agar, raffinose agar, xylose agar, and L. casei agar. Incubations were carried out under aerobic and anaerobic conditions at 27, 30, 37, 43, and 45 degrees C for 24, 72 h, and 7 to 9 d. S. thermophilus agar and aerobic incubation at 37 degrees C for 24 h were suitable for S. thermophilus. L. delbrueckii ssp. bulgaricus could be enumerated using MRS agar (pH 4.58 or pH 5.20) and under anaerobic incubation at 45 degrees C for 72 h. MRS-vancomycine agar and anaerobic incubation at 43 degrees C for 72 h were suitable to enumerate L. rhamnosus. MRS-vancomycine agar and anaerobic incubation at 37 degrees C for 72 h were selective for L. casei. To estimate the counts of L. casei by subtraction method, counts of L. rhamnosus on MRS-vancomycine agar at 43 degrees C for 72 h under anaerobic incubation could be subtracted from total counts of L. casei and L. rhamnosus enumerated on MRS-vancomycine agar at 37 degrees C for 72 h under anaerobic incubation. L. acidophilus could be enumerated using MRS-agar at 43 degrees C for 72 h or Basal agar-maltose agar at 43 degrees C for 72 h or BA-sorbitol agar at 37 degrees C for 72 h, under anaerobic incubation. Bifidobacteria could be enumerated on MRS-NNLP agar under anaerobic incubation at 37 degrees C for 72 h. Propionibacteria could be enumerated on sodium lactate agar under anaerobic incubation at 30 degrees C for 7 to 9 d. A subtraction method was most suitable for counting propionibacteria in the presence of other lactic acid bacteria from a product. For this method, counts of lactic bacteria at d 3 on sodium lactate agar under anaerobic incubation at 30 degrees C were subtracted from counts at d 7 of lactic bacteria and propionibacteria.     

UV-C treatment of juices to inactivate microorganisms using Dean vortex technology

A laboratory-scale UV-C treatment device based on Dean vortex technology was tested for its potential to inactivate spoilage microorganisms in cloudy fruit juices. A log 5 and log 6 reduction could be achieved by inactivating Lactobacillus plantarum BFE 5092 and Escherichia coli DH5α in naturally cloudy apple juice at 1.9 and 7.7 kJ/L, respectively. A treatment with 9.6 kJ/L led to an approximately log 4 inactivation of Saccharomyces cerevisiae DSM 70478 and Alicyclobacillus acidoterrestris DSM 2498. The effects of possible influencing parameters such as optical density, turbidity and viscosity were analyzed with regard to the efficiency of the UV-C treatment. The optical density based on dissolved compounds appeared to be the most important factor which influenced the bacterial inactivation efficiency. Cell counts of L. plantarum BFE 5092 could be reduced in quarter-strength Ringer’s solution adjusted with dye from an initial level of approximately 1 × 10⁸-1 × 10¹ cfu/mL at an optical density (254 nm) of 20 at 9.6 kJ/L. Only a log 1.5 reduction, however, could be achieved at an optical density (254 nm) of 140 using the same UV-C treatment. Furthermore, no noticeable effect on inactivation could be determined by varying the turbidity or the viscosity of the juices investigated. An increasing flow rate and the consequently higher Dean number clearly improved the efficacy of the UV-C treatment. Thus, the inactivation of L. plantarum BFE 5092 in blood orange juice could be enhanced by an approximately 2.5-log reduction by increasing the Dean number from 32 to 256 at 7.7 kJ/L. The UV-C treatment using Dean vortex technology was shown in this study to effectively inactivate microorganisms even in cloudy juices. The optical density value seemed to be the exclusive determining factor on the efficiency of the UV-C inactivation of microorganisms based on Dean vortex technology, while the effect of suspended solids was negligible as a result of the efficient mixing by Dean vortices.     

Clarification of fruit juices by fungal pectinases

This study analyses the efficiency of fungal pectinolytic preparations produced in laboratory and commercial products used in the clarification process of apple, butia palm fruit, blueberry, and grape juices. Two crude enzymatic extracts, produced by Aspergillus niger T0005007-2 (TE1) and Aspergillus oryzae IPT 301 (TE2), were tested in solid-state and submerse cultures, respectively, in comparison to the commercial preparations Pectinex^®Clear and Pectinex^®BE Colour, used for the clarification of clear (apple, butia palm fruit) and dark (blueberry and grape) juices, respectively. With pectinases total activity at 1 U/mL of fruit juice, reactions were conducted at 30 and 50 °C, for 30 and 60 min. Time increase resulted in an improved clarification, whereas temperature increase could not be clearly linked to a greater clarification. When compared to the commercial products, the use of the crude preparation TE1 resulted in similar clarification of apple and blueberry juices. For butia palm and grape juice, however, TE1 presented the highest clarification results, revealing the potential of A. niger T0005007-2 enzymes for commercial application.