Widely distributed lysogeny in probiotic lactobacilli represents a potentially high risk for the fermentative dairy industry

Prophages account for most of the genetic diversity among strains of a given bacterial species, and represent a latent source for the generation of virulent phages. In this work, a set of 30 commercial, collection and dairy-isolated Lactobacillus casei group strains were used. A species-specific PCR assay allowed a reclassification, mainly of strains previously considered Lactobacillus casei, into either Lactobacillus paracasei or Lactobacillus rhamnosus. All the strains were induced with mitomycin C, allowing direct recovering of phage DNA in 25 cases, which corroborates the widely occurrence of lysogeny on Lactobacillus genomes, including probiotic strains of Lactobacillus casei group. Ten out of 11 commercial strains studied contained prophages, evidencing the potential risks of their use at industrial scale. Strains were also induced by treatment with different concentrations of hydrogen peroxide but, however, this agent was not able to evidence a prophage release for any of the strains tested. According to a RAPD-PCR fingerprinting with M13, 1254 and G1 primers, most of the commercial strains presented a high degree of homology and, regarding BglII- and BamHI-restriction profiles of phage DNA, six of them harboured the same prophage. Surprisingly, both Lactobacillus paracasei ATCC 27092 and Lactobacillus paracasei ATCC 27139 shared a second prophage with both an INLAIN collection and a commercial Lactobacillus paracasei strains, whereas two collection strains shared a third one. On the other hand, mitomycin C-inducible prophages were detected only on about a half of the strains isolated from dairy products, which had (with only one exception) from moderate to high correlation coefficients according to RAPD-PCR fingerprinting. After induction, supernatants were filtered and tested against nine Lactobacillus strains of the set sensitive to previously assayed virulent phages, allowing isolation of two new virulent phages: ф iLp1308 and ф iLp84. Both phages were able to lyse all but one strains sensitive to previously assayed phage MLC-A.     

Evaluation of the functional potential of Weissella and Lactobacillus isolates obtained from Nigerian traditional fermented foods and cow's intestine

The characterisation of 24 lactic acid bacteria (LAB) isolates from Nigerian traditional fermented dairy foods, including some cow's intestine isolates, was conducted in order to select isolates for potential use as probiotics. LAB isolates were identified by partial sequencing the 16S rRNA gene as belonging to the species Lactobacillus paracasei, Lactobacillus brevis and mainly Weissella confusa. At the end of a characterisation process, 2 L. paracasei and 2 W. confusa isolates were selected, and their resistance to a simulated gastrointestinal digestion and their ability to adhere to eukaryotic cell lines were assessed. The survival to the simulated gastrointestinal passage was higher when bacterial suspensions were made in skimmed milk (2.0 ± 0.8 log units reduction) or at the simulated gastric juice pH 3 (2.7 ± 0.9 log units reduction) than at pH 2.0 (5.5 ± 0.7 log units reduction). Adhesion of LAB to both intestinal and vaginal epithelial models was comparable or higher than that of the reference Lactobacillus rhamnosus GG. However, some of the isolates increased the adhesion of the pathogen Escherichia coli LMG2092 to HT-29 and HeLa monolayers. Overall, isolates L. paracasei UI14 and W. confusa UI7 are good candidates for further studying potential benefits that support their use as probiotics. This is one of the few articles reporting the characterisation and the probiotic potential of Weissella, although more studies are needed in order to establish their safety for potential probiotic applications.     

Synbiotic potential of fresh cream cheese supplemented with inulin and Lactobacillus paracasei in co-culture with Streptococcus thermophilus

The influence of the addition of Lactobacillus paracasei and Streptococcus thermophilus on the fructan content at the beginning and at the end of storage at 4 ± 1 °C of a potentially synbiotic fresh cream cheese manufactured with inulin was investigated. Three cheese-making trials were prepared, all supplemented with a lactic culture of S. thermophilus (T1, T2 and T3). L. paracasei subsp. paracasei was added in T1 and T2. Inulin was added in T2 and the fructan content was measured after 1 and 21 days of storage. Samples of T2 possessed similar mean concentrations of fructans after 1 and 21 days of storage, 7.32% and 7.27%, respectively, and no significant difference was observed. These results indicated that the metabolism of starter and probiotic bacteria did not degrade the fructans present in those cheeses. Additionally, synbiotic cheeses possessed a fructan content higher than 7 g per 100 g, sufficient to confer prebiotic potential during the entire storage period of these products.     

Kinetic of production and mode of action of the Lactobacillus paracasei subsp. paracasei anti-listerial bacteriocin, an Algerian isolate

Lactobacillus paracasei subsp. paracasei was isolated in our laboratory from breast-fed newborn faeces and identified phenotypically and genotypically. The strain was able to produce a bacteriocin-like substance active towards listerial strains (Listeria innocua CLIP 74915 and Listeria monocytogenes EGDe). The maximum production of the substance by producing strain was detected in the late logarithmic growth phase (14 h in MRS and 18 h in BHI broths). It displayed bactericidal mode of action with leakage of cellular content (K⁺ and ATP) leading to cell lysis as secondary effect. A reduction of about 5 log with 35.7 ± 0.2% of cell lysis and of about 4 log with 82.0 ± 0.3% of cell lysis were observed, respectively, in a phosphate buffer and BHI suspensions. This was further demonstrated by electron microscopy that showed severe modifications in the cell morphology with a concomitant lysis.     

Characterization and adsorption of a Lactobacillus plantarum virulent phage

Bacteriophage infection of lactic acid bacteria is considered one of the biggest worldwide problems in the food industry. Bacteriophages may cause negative effects on the fermentation of various dairy-based products. A virulent bacteriophage was isolated from an abnormal fermentation liquid of Lactobacillus plantarum IMAU10120. The characterization and influence of temperature, pH, divalent cations, and chloramphenicol on the adsorption ability of this phage were evaluated. The results showed that this phage belonged to the Siphoviridae family. It exhibited a burst time of 135 min and a burst size of approximately 215 counts expressed per milliliter per infective center. No significant effect was shown to influence its viability and adsorption at 10 to 37°C. More than 90% of phages exhibited infectivity from pH 5 to 9. Divalent ions and chloramphenicol did not have a significant influence on the adsorption of this phage. The information obtained in this study will enrich the database of lactobacilli virulent phages and provide a basis of information for the control of phages in the food fermentation industry.     

Effect of buckwheat flour and oat bran on growth and cell viability of the probiotic strains Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their combination SYNBIO®, in synbiotic fermented milk

Fermented foods have a great significance since they provide and preserve large quantities of nutritious foods in a wide diversity of flavors, aromas and texture, which enrich the human diet. Originally fermented milks were developed as a means of preserving nutrients and are the most representatives of the category. The first aim of this study was to screen the effect of buckwheat flour and oat bran as prebiotics on the production of probiotic fiber-enriched fermented milks, by investigating the kinetics of acidification of buckwheat flour- and oat bran-supplemented milk fermented by Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their 1:1 combination named SYNBIO®. The probiotic strains viability, pH and sensory characteristics of the fermented fiber-enriched milk products, stored at 4 °C for 28 days were also monitored. The results showed that supplementation of whole milk with the tested probiotic strains and the two vegetable substrates results in a significant faster lowering of the pH. Also, the stability of L. rhamnosus IMC 501®, L. paracasei IMC 502® and SYNBIO® during storage at 4 °C for 28 days in buckwheat flour- and oat bran-supplemented samples was remarkably enhanced. The second aim of the study was to develop a new synbiotic product using the best combination of probiotics and prebiotics by promoting better growth and survival and be acceptable to the consumers with high concentration of probiotic strain. This new product was used to conduct a human feeding trial to validate the fermented milk as a carrier for transporting bacterial cells into the human gastrointestinal tract. The probiotic strains were recovered from fecal samples in 40 out of 40 volunteers fed for 4 weeks one portion per day of synbiotic fermented milk carrying about 10⁹ viable cells.     

Potentially probiotic and synbiotic chocolate mousse

Several studies in recent years have shown the benefits deriving from the ingestion of probiotics, and a large number of products containing lactobacilli and bifidobacteria have been formulated. The purpose of this study was to develop a chocolate mousse to which probiotic and prebiotic ingredients were added, and verify the perspectives of the product with regard to potential for consumer health benefits and sensorial acceptance. Probiotic and synbiotic chocolate mousse supplemented with Lactobacillus paracasei subsp. paracasei LBC 82, solely (P) or together with the prebiotic ingredient inulin (S), were prepared, as well as a control mousse (C). The products were monitored for the population of L. paracasei and contaminants, during storage at 5 °C for up to 28 days, and sensory preference was also tested. Storage trials showed that the viability of the probiotic was retained over 28 days, but the growth of yeasts and moulds might limit the shelf-life of the product. Chocolate mousse was shown to be an excellent vehicle for the delivery of L. paracasei, and the prebiotic ingredient inulin did not interfere in its viability. Moreover, the addition of the probiotic microorganism and of the prebiotic ingredient did not interfere in the sensorial preference of the product.     

Survival and sensory acceptability of probiotic microorganisms in a nonfermented frozen vegetarian dessert

Probiotic microorganisms were incorporated into a nonfermented, vegetarian frozen soy dessert at initial populations greater than 10⁶ cfu/g. The product was assessed for the survival of probiotic microorganisms and sensory acceptability. Lactobacillus acidophilus MJLA1, L. rhamnosus 100-C, L. paracasei ssp. paracasei 01, Bifidobacterium lactis BBDB2, B. lactis BB-12 all survived the 6 month storage trial at populations of 10⁷ cfu/g or greater. Saccharomyces boulardii 74012 did not retain sufficient viability, decreasing below the desirable level of 10⁶ cfu/g. To detect sensory differences, product containing L. acidophilus MJLA1, S. boulardii 74012 and an uninoculated control were stored for 0, 4 and 7 months and compared using triangle tests. Product inoculated with L. acidophilus MJLA1 could not be distinguished from the control sample. Product with S. boulardii 74012 differed from the control and L. acidophilus MJLA1 and developed undesirable flavours during storage. The frozen soy dessert was a suitable food for the delivery of bacterial probiotic strains with excellent viability and acceptable sensory characteristics.     

Development of a locally sustainable functional food based on mutandabota,a traditional food in southern Africa

A probiotic dairy product was developed on the basis of a traditional dish called mutandabota to enable resource-poor populations in southern Africa to benefit from a functional food. Mutandabota is widely consumed in rural southern Africa, making it an ideal food matrix to carry probiotics. First, a process to produce probiotic mutandabota was designed. Raw cow milk was boiled and subsequently cooled to ambient temperature (25°C). Next, dry pulp from the fruit of the baobab tree (Adansonia digitata L.) was added to the milk at a concentration of 4% (wt/vol). This mixture was inoculated with the probiotic Lactobacillus rhamnosus yoba and left to ferment for 24 h, while the growth of the bacterial culture was monitored. Final ingredients were then added to produce probiotic mutandabota that had 14% (wt/vol) baobab fruit pulp and 7% (wt/vol) sugar in cow milk. The pH of probiotic mutandabota was pH 3.5, which ensures that the product is microbiologically safe. The viable plate count of L. rhamnosus yoba increased from 5.8 ± 0.3 log cfu/mL at the point of inoculation to 8.8 ± 0.4 log cfu/mL at the moment of consumption, thereby meeting the criterion to have a viable count of the probiotic bacterium in excess of 6 log cfu/mL of a product. Baobab fruit pulp at 4% promoted growth of L. rhamnosus yoba with a maximal specific growth rate (μ_max) of 0.6 ± 0.2/h at 30°C. The developed technology, though specific for this particular product, has potential to be applied for the delivery of probiotics through a variety of indigenous foods in different regions of the world. Upon consumption, probiotic mutandabota is expected to improve the population's intestinal health, which is especially relevant for vulnerable target groups such as children and elderly people.     

Elaboration of a probiotic oblea from whey fermented using Lactobacillus acidophilus or Bifidobacterium infantis

A novel probiotic product was developed, which was formulated as an oblea (wafer-type dehydrated traditional Mexican dessert) using goat sweet whey fermented with Bifidobacterium infantis or Lactobacillus acidophilus. To obtain the probiotic oblea, the fermented whey was formulated with prebiotic carbohydrates (inulin and resistant starch) and gelatin, and the preparation was poured onto a polytetrafluoroethylene-coated nonstick baking pan, dried in a convection oven, and finally dehydrated at a low relative humidity and room temperature (23 ± 2°C). The amounts of prebiotic carbohydrates and gelatin to be used in the formulation were determined by a factorial experimental design. An untrained sensory panel evaluated 3 quality characteristics (film formation, homogeneity, and smoothness) in the final product. Three different drying temperatures were tested, namely, 40, 55, and 70°C. Bacterial survival at each temperature was determined by viable plate-counting. The best formulation, based on the quality characteristics tested, consisted of 58.33% (vol/vol) of fermented whey, 8.33% (vol/vol) of 6% (wt/vol) resistant starch dispersion, 16.66% (vol/vol) of 15% (wt/vol) inulin solution, and 16.66% (vol/vol) of a 10% (wt/vol) gelatin solution. Drying at 55 ± 2°C for 2.66 ± 0.22 h allowed for concentrations of probiotic bacteria above 9 log₁₀ cfu/g, which is above the minimum concentration required in a probiotic product.     

Growth of Lactobacillus paracasei ATCC 334 in a cheese model system: a biochemical approach

Growth of Lactobacillus paracasei ATCC 334, in a cheese-ripening model system based upon a medium prepared from ripening Cheddar cheese extract (CCE) was evaluated. Lactobacillus paracasei ATCC 334 grows in CCE made from cheese ripened for 2 (2mCCE), 6 (6mCCE), and 8 (8mCCE) mo, to final cell densities of 5.9 × 10⁸, 1.2 × 10⁸, and 2.1 × 10⁷ cfu/mL, respectively. Biochemical analysis and mass balance equations were used to determine substrate consumption patterns and products formed in 2mCCE. The products formed included formate, acetate, and d-lactate. These data allowed us to identify the pathways likely used and to initiate metabolic flux analysis. The production of volatiles during growth of Lb. paracasei ATCC 334 in 8mCCE was monitored to evaluate the metabolic pathways utilized by Lb. paracasei during the later stages of ripening Cheddar cheese. The 2 volatiles detected at high levels were ethanol and acetate. The remaining detected volatiles are present in significantly lower amounts and likely result from amino acid, pyruvate, and acetyl-coenzyme A metabolism. Carbon balance of galactose, lactose, citrate, and phosphoserine/phosphoserine-containing peptides in terms of d-lactate, acetate, and formate are in agreement with the amounts of substrates observed in 2mCCE; however, this was not the case for 6mCCE and 8mCCE, suggesting that additional energy sources are utilized during growth of Lb. paracasei ATCC 334 in these CCE. This study provides valuable information on the biochemistry and physiology of Lb. paracasei ATCC 334 in ripening cheese.     

Effect of exopolysaccharides (EPSs) produced by Lactobacillus delbrueckii subsp. bulgaricus strains to bacteriophage and nisin sensitivity of the bacteria

Lactobacillus strains used in this study were isolated from village-type yogurt and raw milk. The isolates were identified as Lactobacillus delbrueckii subsp. bulgaricus by 16 s rDNA sequence analysis and API 50 CHL identification systems. The exopolysaccharide (EPS) production of the strains growth in skim milk were investigated. In addition sensitivity and insensitivity of these strains against domestic bacteriophages and nisin were examined. It was deduced that those strains which had relatively high EPS-producing capacity were insensitive against phages and nisin. Linear relationships were determined between EPS production of the bacteria and bacteriophage and nisin insensitivity of the bacteria.There was a negative correlation between EPS production quantity and phage and nisin sensitivity of the bacteria. Of all the strains, L. delbrueckii subsp bulgaricus B3 produced the highest EPS quantity, and it was insensitive against phages and nisin. Based on these results, it is suggested that L. delbrueckii subsp bulgaricus B3 can be used with the starter culture in dairy industry for stable and high-quality yogurt production.     

Temperate and virulent Lactobacillus delbrueckii bacteriophages: Comparison of their thermal and chemical resistance

The aim of this work was to study the efficiency of diverse chemical and thermal treatments usually used in dairy industries to control the number of virulent and temperate Lactobacillus delbrueckii bacteriophages. Two temperate (Cb1/204 and Cb1/342) and three virulent (BYM, YAB and Ib3) phages were studied. The thermal treatments applied were: 63 °C for 30 min (low temperature – long time, LTLT), 72 °C for 15 s (high temperature – short time, HTST), 82 °C for 5 min (milk destined to yogurt elaboration) and 90 °C for 15 min (FIL-IDF). The chemical agents studied were: sodium hypochlorite, ethanol, isopropanol, peracetic acid, biocides A (quaternary ammonium chloride), B (hydrogen peroxide, peracetic acid and peroctanoic acid), C (alkaline chloride foam), D (p-toluensulfonchloroamide, sodium salt) and E (ethoxylated nonylphenol and phosphoric acid). The kinetics of inactivation were drew and T₉₉ (time necessary to eliminate the 99% of phage particles) calculated. Results obtained showed that temperate phages revealed lower resistance than the virulent ones to the treatment temperatures. Biocides A, C, E and peracetic acid showed a notable efficiency to inactivate high concentrations of temperate and virulent L. delbrueckii phages. Biocide B evidenced, in general, a good capacity to eliminate the phage particles. Particularly for this biocide virulent phage Ib3 showed the highest resistance in comparison to the rest of temperate and virulent ones. On the contrary, biocide D and isopropanol presented a very low capacity to inactivate all phages studied. The efficiency of ethanol and hypochlorite was variable depending to the phages considered. These results allow a better knowledge and give useful information to outline more effective treatments to reduce the phage infections in dairy plants.     

Effects of a probiotic fermented milk beverage containing Lactobacillus casei strain Shirota on defecation frequency,intestinal microbiota,and the intestinal environment of healthy individuals with soft stools

The effects of drinking a fermented milk beverage that contains Lactobacillus casei strain Shirota (LcS) at 40 billion bacterial cells/bottle for 4 weeks (probiotics, 1 bottle/day) on defecation frequency, intestinal microbiota and the intestinal environment of healthy individuals with soft stools were evaluated. Thirty-four healthy adults who had soft stools were randomised into 2 groups, and the effects of a regular 4-week intake of probiotics were evaluated by a placebo-controlled, double-blind, parallel-group comparative design. Defecation frequency significantly decreased after the 4-week intake period compared with before the probiotic treatment. The stool quality significantly improved (hardened) compared to the placebo. Also, the water content of the stools was lower in the probiotic group than in the placebo group. Live LcS was recovered at 6.9 ± 1.3 and 7.2 ± 0.8 log₁₀ CFU per 1 g of stool after 2 and 4 weeks, respectively, of probiotic treatment. The number of bifidobacteria in the stools also increased significantly compared with the level before starting the probiotics. The organic acid levels (total, acetic acid, propionic acid, and butyric acid) significantly increased compared with the level before intake in both the probiotic and placebo groups, but they returned to the original levels after the end of the intake period. These results suggest that probiotic fermented milk beverage has an intestine-conditioning effect by improving the frequency of defecation and stool quality and increasing the intrinsic bifidobacteria in healthy individuals with soft stool.     

Effect of calcium alginate and resistant starch microencapsulation on the survival rate of Lactobacillus acidophilus La5 and sensory properties in Iranian white brined cheese

Two types of probiotic cheese, with free and microencapsulated bacteria, were manufactured in triplicate under the same conditions. The number of viable cells during 182 days of storage in refrigerated conditions was evaluated. The number of viable cells of Lactobacillus acidophilus was reduced significantly from day 28 to day 182 of storage period in both types of cheese, but reduction in the cheese containing free cells (5.1 ± 0.67 log cfu g⁻¹) was significantly p < 0.05 higher than the cheese containing microencapsulated cells (11.00 ± 0.58 log cfu g⁻¹). The results showed that, microencapsulation in calcium alginate gel and resistant starch was able to increase the survival rate of L. acidophilus La5 in Iranian white brined cheese after 6 months of storage.     

Survival of probiotic adjunct cultures in cheese and challenges in their enumeration using selective media

Various selective media for enumerating probiotic and cheese cultures were screened, with 6 media then used to study survival of probiotic bacteria in full-fat and low-fat Cheddar cheese. Commercial strains of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, or Bifidobacterium lactis were added as probiotic adjuncts. The selective media, designed to promote growth of certain lactic acid bacteria (LAB) over others or to differentiate between LAB, were used to detect individual LAB types during cheese storage. Commercial strains of Lactococcus, Lactobacillus, and Bifidobacterium spp. were initially screened on the 6 selective media along with nonstarter LAB (NSLAB) isolates. The microbial flora of the cheeses was analyzed during 9 mo of storage at 6°C. Many NSLAB were able to grow on media presumed selective for Lactococcus, Bifidobacterium spp., or Lb. acidophilus, which became apparent after 90 d of cheese storage, Between 90 and 120 d of storage, bacterial counts changed on media selective for Bifidobacterium spp., suggesting growth of NSLAB. Appearance of NSLAB on Lb. casei selective media [de man, Rogosa, and Sharpe (MRS) + vancomycin] occurred sooner (30 d) in low-fat cheese than in full-fat control cheeses. Differentiation between NSLAB and Lactococcus was achieved by counting after 18 to 24 h when the NSLAB colonies were only pinpoint in size. Growth of NSLAB on the various selective media during aging means that probiotic adjunct cultures added during cheesemaking can only be enumerated with confidence on selective media for up to 3 or 4 mo. After this time, growth of NSLAB obfuscates enumeration of probiotic adjuncts. When adjunct Lb. casei or Lb. paracasei cultures are added during cheesemaking, they appear to remain at high numbers for a long time (9 mo) when counted on MRS + vancomycin medium, but a reasonable probability exists that they have been overtaken by NSLAB, which also grow readily on this medium. Enumeration using multiple selective media can provide insight into whether it is the actual adjunct culture or a NSLAB strain that is being enumerated.     

Isolation and characterisation of a novel Podoviridae-phage infecting Weissella cibaria N 22 from Nham, a Thai fermented pork sausage

A novel Podoviridae lactic acid bacteria (LAB) phage from Nham, a Thai fermented pork sausage, is reported. From a total of 36 samples, 41 isolates of LAB were obtained and employed as hosts for the isolation of phages. From these LAB, only one phage, designated Φ 22, was isolated. The lactic acid bacterial isolate named N 22, sensitive to phage Φ 22 infection was identified by an API 50 CHL kit and N 22’s complete sequence of the 16S rDNA sequence. BLASTN analysis of the 16S rDNA sequence revealed a 99% similarity to the 16S rDNA sequence of Weissella cibaria in the GenBank database. Electron micrographs indicated that the phage head was icosahedral with head size and tail length of 92 × 50 nm and 27 nm, respectively. On the basis of the morphology, this phage belongs to the family Podoviridae. Host-range determination revealed that the phage Φ 22 was not capable of infecting the other 40 isolates of LAB and referenced Weissella strains used. A one-step growth experiment showed that the latent period and burst size were estimated at 110 min and 55 phage particles/infected cell, respectively. Furthermore, the phage was infective over a wide range of pH (pH 5.0-8.0) and the D time of Φ 22 was calculated as 88 s at 70 °C and 15 s at 80 °C. Phage titers decreased below the detection limit (20 PFU/ml) after heating for more than 60 s at 80 °C, or 20 s at 90 °C or less than 10 s at 100 °C. The results from the study of Nham revealed that Φ 22 was active against the potential starter culture (W. cibaria N 22) for Nham fermentation. Phage infection could adversely affect the fermentation process of Nham by delaying acidification when using W. cibaria N 22 as a starter. However, the results from a sensory test revealed that the panelists did not detect any defects in the final products. This is the first report on the isolation of W. cibaria phage.     

Genome analysis of Lactobacillus fermentum temperate bacteriophage ФPYB5

The complete genomic sequence of Lactobacillus fermentum temperate bacteriophage ФPYB5 was determined. The phage possesses a linear, double-stranded DNA genome of 32,847 bp with a G + C content of 45.21%. A total of 46 putative open reading frames (ORFs) were identified. On the basis of homology comparisons, 25 ORFs could be assigned putative functions. The genome of bacteriophage ФPYB5 is highly modular with functionally related genes clustered together. Genome DNA of temperate bacteriophage ФPYB5, induced from heterofermentative lactic acid bacteria, showed to be closely related to that of the prophage of heterofermentative Lactobacillus reuteri 100-23 and heterofermetative Leuconostoc oenos bacteriophage 10MC in an evolutionary aspect.     

Characterisation of three temperate phages released from the same Lactobacillus paracasei commercial strain

Lysogeny entails more economical and technological risks in probiotic Lactobacillus casei/paracasei bacteria than in other lactic acid bacteria, due to economic value. Lysogeny is widely spread among L. casei/paracasei strains. Siphophages C_L1 and C_L2, isolated from noninfected lysed‐cultures of commercial L. paracasei A, are thermoresistant, have identical host spectrum, latent and burst times, whereas burst sizes are 148 and 85, respectively. Mitomycin C induction of L. paracasei A yielded prophage iA2, whose presence was confirmed on the strain genome and whose restriction patterns differed from phages C_L1 and C_L2. The latter shared several restriction fragments, probably indicating a common origin.     

Effect of high pressure homogenization on lactic acid bacteria phages and probiotic bacteria phages

To investigate the effect of high pressure homogenization on virus inactivation, phages specific for Lactobacillus delbrueckii, Lactobacillus helveticus, Streptococcus thermophilus, Lactococcus lactis, Lactobacillus paracasei and Lactobacillus plantarum were studied. The influence of pressure, number of passes, suspension medium and phage concentration were studied at 25 °C. Reductions in viability were proportional to pressure and number of passes, though the inactivation extent was phage-dependent. At 100 MPa, some bacteriophages were completely inactivated (6 log₁₀ reduction) after 3 or 5 passes, while others remained infective after 8 passes. For all phages, treatment at 60 MPa was insufficient for complete inactivation, even after 8 passes. No clear influence of suspending medium was observed. Inactivation seems to depend on phage concentration; the higher the initial load, the bigger the reduction achieved. Although these results showed that several phages studied are resistant to high-pressure homogenization, this strategy could be combined with others to control their presence in raw milk.