Effect of combinations of high-pressure treatment and bacteriocin-producing lactic acid bacteria on the survival of Listeria monocytogenes in raw milk cheese

The combined effect of high-pressure (HP) treatment and bacteriocin-producing lactic acid bacteria (BP-LAB) on the survival of Listeria monocytogenes Scott A in cheeses made from raw milk that was inoculated with the pathogen at 4.80 log cfu mL⁻¹, a commercial starter and one of seven strains of BP-LAB was investigated. On day 3, the counts of L. monocytogenes were 7.03 log cfu g⁻¹ in a control cheese (without BP-LAB, not HP treated), 6.06–6.74 log cfu g⁻¹ in cheeses with BP-LAB, 6.13 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 300 MPa, 2.01 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 500 MPa, 3.83–5.43 log cfu g⁻¹ in cheeses with BP-LAB and treated on day 2 at 300 MPa, and 1.81 log cfu g⁻¹ or less in cheeses with BP-LAB and treated on day 2 at 500 MPa. HP treatment was more effective on day 51 than on day 2.     

Survival of Listeria monocytogenes in Galotyri,a traditional Greek soft acid-curd cheese,stored aerobically at 4°C and 12°C

Galotyri is a traditional Greek soft acid-curd cheese, which is made from ewes’ or goats’ milk and is consumed fresh. Because cheese processing may allow Listeria monocytogenes post-process contamination, this study evaluated survival of the pathogen in fresh cheese during storage. Portions (0.5 kg) of two commercial types (<2% salt) of Galotyri, one artisan (pH 4.0±0.1) and the other industrial (pH 3.8±0.1), were inoculated with ca. 3 or 7 log cfu g⁻¹ of a five-strain cocktail of L. monocytogenes and stored aerobically at 4°C and 12°C. After 3 days, average declines of pathogen's populations (PALCAM agar) were 1.3–1.6 and 3.7–4.6 log cfu g⁻¹ in cheese samples for the low and high inocula, respectively. These declines were independent (P>0.05) of the cheese type or the storage temperature. From day 3, however, declines shifted to small or minimal to result in 1.4–1.8 log cfu g⁻¹ of survivors at 28 days of storage of all cheeses at 4°C, indicating a strong “tailing” independent of initial level of contamination. Low (1.2–1.7 log cfu g⁻¹) survival of L. monocytogenes also occurred in cheeses at 12°C for 14 days, which were prone to surface yeast spoilage. When ca. 3 log cfu g⁻¹ of L. monocytogenes were inoculated in laboratory scale prepared Galotyri of pH ≅4.4 and ≅3% salt, the pathogen died off at 14 and 21 days at 12°C and 4°C, respectively, in artisan type cheeses fermented with the natural starter. In contrast, the pathogen survived for 28 days in cheeses fermented with the industrial starter. These results indicate that L. monocytogenes cannot grow but may survive during retail storage of Galotyri despite its low pH of or slightly below 4.0. Although contamination of Galotyri with L. monocytogenes may be expected low (<100 cfu g⁻¹) in practice, that long-term survival of the pathogen in commercial cheeses was shown to be unaffected by the artificial contamination level (3 or 7 logs) and the storage temperature (4°C or 12°C), which should be a concern.     

A combined morphologic and molecular approach for characterizing fungal microflora from a traditional Italian cheese (Fossa cheese)

Phenotypic and genotypic methods were used to identify filamentous fungi that characterize traditional Italian Fossa cheese and its ripening environment. After ageing for 60 days at a dairy, it was ripened for an additional three months in a pit. In the fully ripened cheese, moulds ranged from 3 to 3.4 log cfu g^?1 and Penicillium was the prevalent species. Pit environmental fungi ranged from 530 to 750 cfu m^?3 (air) and from 130 to 340 cfu cm^?2 (surfaces). The dominant pit strains were Alternaria spp., Aspergillus spp., Cladosporium spp. and Penicillium spp. Phylogenetic analyses of 18S rRNA gene and ITS1-5.8S-ITS2 regions highlighted Penicillium camemberti, Aspergillus nidulans and Aspergillus versicolor as traceable species occurring in both the cheese and pit environment, suggesting their involvement in the development of typical Fossa cheese characteristics. This approach may be used for the identification of microflora on other cheese varieties to better understand the fungal contribution in cheese ripening.     

Production of pediocin PA-1, and coproduction of nisin A and pediocin PA-1, by wild Lactococcus lactis strains of dairy origin

Heterologous production of pediocin PA-1 in nisin and non-nisin-producing Lactococcus lactis strains, which had been previously selected because of their technological properties for cheese making, was investigated. Plasmid pFI2160, which contains a hybrid gene (L-pedA) encoding the fusion between the lactococcin A leader and propediocin PA-1, and also the genes lcnC and lcnD, that encode the lactococcin A secretion apparatus, was introduced into L. lactis ESI 153 and L. lactis ESI 515 (Nis⁺). The pediocin production level of their respective transformants, L. lactis CL1 and L. lactis CL2 (Nis⁺), was approximately 600 and 400 ng mL⁻¹, respectively, which represents a 30% and a 20% of the quantity produced by the natural pediocin PA-1 producer Pediococcus acidilactici 347. Transformation of L. lactis ESI 515 with pFI2160 did not affect its ability to produce nisin. Pediocin bioassays showed the stability of pFI2160 in both heterologous hosts under selective and non-selective conditions.     

Fate of enterotoxigenic Staphylococcus aureus in the presence of nisin-producing Lactococcus lactis strain during manufacture of Jben,a Moroccan traditional fresh cheese

The inhibitory effect of nisin-producing Lactococcus lactis subsp. lactis UL730 on the growth of enterotoxigenic Staphylococcus aureus J10 during manufacture of Jben, a Moroccan traditional fresh cheese prepared from recombined milk, was investigated.With an inoculum level of 10³ cfu mL⁻¹, S. aureus was absent in Jben four days after inoculation when the nisin-producing lactococcus was used as lactic starter. In contrast, it survived after that period, when the starter was non-nisin-producing. No staphylococcal thermonuclease was detected in all Jben samples made from milk inoculated with S. aureus at the level of 10³ cfu mL⁻¹.With a higher inoculum of 10⁵ cfu mL⁻¹, S. aureus was still present in Jben after manufacture and persisted during the storage of the product for 3 days in the laboratory, even when the starter used was nisin-producing. Staphylococcal thermonuclease and type C enterotoxin were detected in all Jben samples made from milk inoculated with 10⁵ cfu mL⁻¹. Thermonuclease and enterotoxin were already produced in the coagulum, at 24 h after milk inoculation with S. aureus.     

Addition of probiotic bacteria in a semi-hard goat cheese (coalho): Survival to simulated gastrointestinal conditions and inhibitory effect against pathogenic bacteria

In this study, the survival of the probiotics Lactobacillus acidophilus (LA-5), Lactobacillus casei subsp. paracasei (L. casei 01) and Bifidobacterium lactis (BB12) incorporated in a Brazilian semi-hard goat cheese (coalho) when exposed to in vitro simulated conditions of digestion was assessed. The inhibitory effects of these probiotic bacteria were also evaluated against Listeria monocytogenes and Staphylococcus aureus in the goat coalho cheese during refrigerated storage. At the end of the in vitro digestion, all of the probiotic tested strains presented decreased (p < 0.05) viable cell counts (5.5–6.0 log cfu/g) with respect to those determined before exposure to the mouth conditions (7–8 log cfu/g). L. casei subsp. paracasei presented inhibition rate of 7.87% and 23.63% against S. aureus on the 14th and 21st day of storage at 10 °C, respectively; against L. monocytogenes these values were 12.96 and 32.99%. Positive inhibition rates of B. lactis toward S. aureus were found on the 1st, 14th and 21st days of storage (16.32%, 10.12% and 3.67%, respectively); and against L. monocytogenes only on the 1st day of storage (3.28%). From these results, goat coalho cheese could be an interesting carrier of probiotic strains of L. acidophilus, L. casei subsp. paracasei and B. lactis. Moreover, L. casei subsp. paracasei, could be used as protective culture for delaying the growth of S. aureus and L. monocytogenes in goat coalho cheese.     

Fresh-cheesemilk formulation fermented by a combination of freeze-dried citrate-positive cultures and exopolysaccharide-producing lactobacilli with liquid lactococcal starters

Milk formulation (4% fat and 5% protein) prepared to simulate fresh cheese production was inoculated with: (1) 10⁷ cfu mL⁻¹ of fresh liquid starters of Lactococcus lactis ssp. lactis T1 and Lc. lactis ssp. cremoris T2, (2) a freeze-dried exopolysaccharide-producing (EPS) strain of Lactobacillus rhamnosus RW-9595M, and (3) freeze-dried Leuconostoc cremoris LM057 or Lc. lactis ssp. lactis var. diacetylactis MD089 strains. The effect of inoculation rate of the freeze-dried starters (between 10⁶ and 10⁷ cfu mL⁻¹) and incubation temperature (between 23.5 and 36.5 °C) on evolution of pH and the various populations during fermentation was examined. Texture (apparent viscosity, syneresis potential) and chemical composition (diacetyl, acetaldehyde) of the fermented milks were also determined. Milk was incubated until a pH of 4.6 was obtained, which required between 6 and 10 h depending on temperature.In the range of inoculation levels used, there was no significant effect of the presence of lactobacilli, Ln. cremoris or Lc. lactis ssp. lactis var. diacetilactis on the growth of the lactococci. There was a direct correlation between the inoculation rates of the freeze-dried cultures and their final populations in the fermented milks. The growth of the cultures were also affected by temperature, Ln. cremoris growing less as incubation temperature increased, while the opposite was noted with Lb. rhamnosus. The apparent viscosity of the fermented milk was significantly affected by incubation temperature, but there was no correlation between apparent viscosity and the final population in lactobacilli. Of the three variables studied, the highest correlation with diacetyl content was obtained with the inoculation level of the Leuconostoc strain.     

Enterobacteriaceae in beef products from retail outlets in the Republic of Ireland and comparison of the presence and counts of E. coli O157:H7 in these products

This study investigated the prevalence and numbers of Enterobacteriaceae in minced beef and beef burgers purchased from supermarkets and butcher shops in the Republic of Ireland (RoI). Samples (n=1303) collected between June 2001 and April 2002 from every county in the RoI (∼60 per county) were examined for the presence of Enterobacteriaceae using method BS 5763. Overall, in the 43 beef products in which E. coli O157:H7 was present the Enterobacteriaceae counts ranged from 0.52 to 6.98 log₁₀ cfu g⁻¹. There was no correlation between the number of Enterobacteriaceae and the presence of E. coli O157:H7. There were no significant differences between Enterobacteriaceae numbers in fresh, unpackaged, minced beef samples from butcher shops and supermarkets, or in fresh, unpackaged, beef burgers from butcher shops and supermarkets. However, there were significant differences among the numbers of Enterobacteriaceae detected in different minced beef products. The numbers of Enterobacteriaceae in fresh, unpackaged, minced beef (6.54–6.98 log₁₀ cfu g⁻¹) were considerably higher than in preprepared or prepackaged minced beef (2.95–3.62 log₁₀ cfu g⁻¹).     

Proteolytic pattern and organic acid profiles of probiotic Cheddar cheese as influenced by probiotic strains of Lactobacillus acidophilus,Lb. paracasei,Lb. casei or Bifidobacterium sp.

Cheddar cheeses were produced with starter lactococci and Bifidobacterium longum 1941, B. lactis LAFTI^® B94, Lactobacillus casei 279, Lb. paracasei LAFTI^® L26, Lb. acidophilus 4962 or Lb. acidophilus LAFTI^® L10 to study the survival of the probiotic bacteria and the influence of these organisms on proteolytic patterns and production of organic acid during ripening period of 6 months at 4 °C. All probiotic adjuncts survived the manufacturing process of Cheddar cheese at high levels without alteration to the cheese-making process. After 6 months of ripening, cheeses maintained the level of probiotic organisms at >8.0 log₁₀ cfu g⁻¹ with minimal effect on moisture, fat, protein and salt content. Acetic acid concentration was higher in cheeses with B. longum 1941, B. lactis LAFTI^® B94, Lb. casei 279 and Lb. paracasei LAFTI^® L26. Each probiotic organism influenced the proteolytic pattern of Cheddar cheese in different ways. Lb. casei 279 and Lb. paracasei LAFTI^® L26 showed higher hydrolysis of casein. Higher concentrations of free amino acids (FAAs) were found in all probiotic cheeses. Although Bifidobacterium sp. was found to be weakly proteolytic, cheeses with the addition of those strains had highest concentration of FAAs. These data thus suggested that Lb. acidophilus 4962, Lb. casei 279, B. longum 1941, Lb. acidophilus LAFTI^® L10, Lb. paracasei LAFTI^® L26 and B. lactis LAFTI^® B94 can be applied successfully in Cheddar cheese.     

Effect of commercial adjunct cultures on proteolysis in low-fat Kefalograviera-type cheese

The effect of two commercially available adjunct cultures, LBC 80 (Lactobacillus casei subsp. rhamnosus) and CR-213 (containing Lactococcus lactis subsp. cremoris and Lc. lactis subsp. lactis) on the proteolysis in low-fat hard ewes’ milk cheese of Kefalograviera-type was investigated. Two controls, a full-fat cheese (306 g kg⁻¹ fat, 378 g kg⁻¹ moisture) and a low-fat cheese (97 g kg⁻¹ fat, 486 g kg⁻¹ moisture, made using a modified procedure), were also prepared. The effect of adjunct culture on proteolysis, as examined by polyacrylamide gel electrophoresis of cheese and water soluble cheese extracts, was marginal. The reverse-phase HPLC peptide profiles of the water soluble extracts from low-fat cheeses were similar although some quantitative differences were observed between low-fat control cheese and experimental cheeses. The fat content as reflected by the differences in peptide profiles affected the pattern of proteolysis. Proteolysis, as measured by the percentage of total nitrogen soluble in water or in 120 g L⁻¹ trichloroacetic acid, was significantly (P<0.05) affected by the addition of adjunct cultures. Furthermore, the adjunct cultures enhanced the production of low molecular mass nitrogenous compounds; the levels of total nitrogen, soluble in 50 g L⁻¹ phosphotungstic acid, and of free amino acids were significantly (P<0.05) higher in the low-fat experimental cheeses than in the low-fat control cheese.     

Probiotic properties of folate producing Streptococcus thermophilus strains

This study aimed at assessing the probiotic potential of two high folate producing Streptococcus thermophilus strains (RD102 and RD104) isolated from Indian fermented milk products by both in vitro and in vivo tests. These strains were able to survive at pH 2.5 and 2% bile with a good bile salt hydrolase activity, cell surface hydrophobicity and sensitivity to most of the clinically important antibiotics. On evaluation for gastrointestinal transit tolerance these showed a viable count of 5 log cfu mL^?1 and 7 log cfu mL^?1, respectively in simulated gastrointestinal juice of pH 2.0 and 2% bile. During the in vivo feeding trial in mice the strains showed a viable count of about 7 log cfu g^?1 faeces and 6 log cfu g^?1 of large intestine, respectively. These strains were hence observed to possess favorable strain specific probiotic properties and have the potential to be a source of novel probiotics.     

Microbiological conditions of moisture-enhanced pork before and after cooking

Substantial numbers of aerobic bacteria but few coliforms or Listeria spp. and no Escherichia coli were recovered from both swab samples and brines circulated in cleaned equipment used for injecting pork loins. After meat was processed for 30 or 60 min, the numbers of aerobic bacteria in brines had increased by >1 log unit, to about 4.5 log cfu ml⁻¹, but coliforms were <2 and E. coli and Listeria spp. were <1 log cfu ml⁻¹. The numbers of bacteria on the surfaces of pork loins before and after injection of the meat were similar. No bacteria were recovered from the deep tissues of the uninjected meat, but aerobic bacteria were recovered at log-mean numbers of 2.1 log cfu g⁻¹ and coliforms at log-total numbers of 1.2 log cfu 25 g⁻¹ from 25 samples of deep tissues of injected meat. Aerobic bacteria were recovered at log total numbers of 1.0 log cfu 25 g⁻¹ from 25 samples of injected pork cooked to a central temperature of 61 °C, but no bacteria were recovered from the deep tissues of meat cooked to 70 °C. The findings suggest that moisture-enhanced pork cooked to a medium rare condition can be microbiologically safe.     

The growth and survival of Escherichia coli O157:H7 on minced bison and pieces of bison meat stored at 5 and 10 °C

This study investigated the growth and survival of Escherichia coli O157:H7 on minced and whole pieces of bison meat. Growth curves of native microflora, including Pseudomonas spp. and Enterobacteriaceae were also generated. A marked E. coli O157:H7 strain was inoculated onto minced and whole pieces of bison meat at an initial level of 1.5 log₁₀ cfu g⁻¹. The inoculated meat was stored at either 5 °C for 28 days or 10 °C for 21 days. Survival, but no growth, of E. coli O157:H7 was observed on both forms of bison meat stored at 5 °C, while significant growth of the organism was observed at 10 °C. E. coli O157:H7 counts on whole pieces were generally higher than counts observed on minced bison meat, and reached their highest population by 14 days, with a total increase of 3.36 log₁₀ cfu g⁻¹ on whole pieces and 2.12 log₁₀ cfu g⁻¹on minced bison meat stored at 10 °C. Under the same storage temperature, Pseudomonas spp. and total counts displayed similar growth patterns on both pieces and minced bison meat, while the Enterobacteriaceae showed a slower growth rate. This study showed that the growth of E. coli O157:H7 on bison meat is similar to that observed in studies of beef.     

Relating physicochemical and microbiological safety indicators during processing of linguiça,a Portuguese traditional dry-fermented sausage

Linguiça is a Portuguese traditional fermented sausage whose microbiological quality and safety can be highly variable. In order to elucidate risk factors and the particularities of the manufacturing technology that explain the between-batch variability in total viable counts (TVC), Enterobacteriaceae, Staphylococcus aureus and Listeria monocytogenes in the product; microbiological and physicochemical characterisation of linguiça at five stages of production (i.e., raw pork meat, mixed with ingredients, macerated, smoked and ripened) was carried out. A total of six production batches were surveyed from two factories; one utilised curing salts and polyphosphate in their formulation (Factory II). The delayed fermentation in the nitrite-formulated sausages was partly responsible for the increase (p < 0.01) in Enterobacteriaceae, S. aureus and L. monocytogenes from raw meat (3.21 log CFU/g, 1.30 log CFU/g and 22.2 CFU/g, respectively) to the end of maceration (4.14 log CFU/g, 2.10 log CFU/g and 140 CFU/g, respectively) while the better acidification process in the nitrite-free sausages (Factory I) led to lower counts of S. aureus (2.64 log CFU/g) and L. monocytogenes (10 CFU/g) in the finished products. In Factory II, although L. monocytogenes entered the chain at the point of mixing, it became steadily inactivated during smoking and ripening (< 50 CFU/g), despite the initially-delayed fermentation. Nitrite had a strong effect on reducing Enterobacteriaceae throughout smoking (r = − 0.73) and ripening (r = − 0.59), while it failed to control the growth of S. aureus. The main hurdle preventing the development of S. aureus in linguiça is the pH, and other factors contributing to its control are: longer ripening days (p = 0.019), low S. aureus in raw meat (p = 0.098), properly-washed casings (p = 0.094), and less contamination during mixing (p = 0.199). In the case of L. monocytogenes, at least three hurdles hinder its development in linguiça: low a_w (p = 0.004), low pH (p = 0.040) and nitrite (p = 0.060), and other factors contributing to its control are: longer ripening (p = 0.072) and maceration (p = 0.106) periods, lower a_w at the end of smoking (p = 0.076) and properly-washed casings (p = 0.099). Results have shown that there is a need to standardise the productive process of linguiça, to optimise the initial acidification process, and to reinforce proper programmes of quality control of ingredients and good hygiene practices, so as to minimise the introduction of Enterobacteriaceae and pathogens from external sources.     

Effect of acidification on the activity of probiotics in yoghurt during cold storage

The viability of Lactobacillus acidophilus LAFTI^® L10, Bifidobacterium lactis LAFTI^® B94, and L. paracasei LAFTI^® L26 and their proteolytic activities were assessed in yoghurt at different termination pH of 4.45, 4.50, 4.55, and 4.60 in the presence of L. delbrueckii ssp. bulgaricus Lb1466 and Streptococcus thermophilus St1342 during 28 days of storage at 4 °C. All strains achieved the recommended level of 6.00 log cfu g⁻¹ of the product with L. acidophilus LAFTI^® L10 and L. paracasei LAFTI^® L26 exceeding the number to 8.00 and 7.00 log cfu g⁻¹, respectively. Lactobacilli strains showed a good cellular stability maintaining constant concentration throughout storage period regardless of termination pH. On the other hand, the cell counts of B. lactis LAFTI^® B94 decreased by one log cycle at the end of storage. The presence of probiotic organisms enhanced proteolysis significantly in comparison with the control batch containing L. delbrueckii ssp. bulgaricus Lb1466 and S. thermophilus St1342 only. The proteolytic activity varied due to termination pH, but also appeared to be strain related. The increased proteolysis improved survival of L. delbrueckii ssp. bulgaricus Lb1466 during storage resulting in lowering of pH and production of higher levels of organic acids, which might have caused the low cell counts for B. lactis LAFTI^® B94.     

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes in milk by caprylic acid and monocaprylin

Listeria monocytogenes and Escherichia coli O157:H7 are major foodborne pathogens implicated in various outbreaks involving pasteurized or unpasteurized milk, and various dairy products. The objective of this study was to determine the antibacterial effect of caprylic acid (CA, C_8:0) and its monoglyceride, monocaprylin (MC) on L. monocytogenes and E. coli O157:H7 in whole milk. A five-strain mixture of E. coli O157:H7 or L. monocytogenes was inoculated in autoclaved milk (10⁶ CFU/ml) containing 0, 25, or 50 mM of CA or MC. At 37°C, all the treatments, excepting 25 mm CA, reduced the population of both pathogens by approximately 5.0 log CFU/ml in 6 h. At 24 h of storage at 8°C, MC at both levels and CA at 50 mM decreased L. monocytogenes and E. coli O157:H7, respectively by >5.0 log CFU/ml. At 48 h of 4°C storage, populations of L. monocytogenes and E. coli O157:H7 were decreased to below detection level (enrichment negative) by 50 mm of MC and CA, respectively. Results indicate that MC could potentially be used to inhibit L. monocytogenes and E. coli O157:H7 in milk and dairy products, but sensory studies need to be conducted before recommending their use.     

Efficacy of plant extracts in inhibitingAeromonas hydrophilaandListeria monocytogenesin refrigerated,cooked poultry

Alcohol extracts of angelica root, banana purée, bay, caraway seed, carrot root, clove (eugenol), marjoram, pimento leaf and thyme were applied to cooked chicken to determine their antimicrobial activities against Aeromonas hydrophilaand Listeria monocytogenes.Skinless chicken breast meat was cooked to an internal temperature of 85°C, allowed to cool to c. 5°C, then treated by surface application with plant extracts. Low (10 cfu g⁻¹)or high (10⁵ cfu g⁻¹)populations of A. hydrophilaand L. monocytogeneswere applied and samples were stored at either 5 or 15°C for up to 14 or 7 days, respectively. Eugenol and pimento extracts were most effective in inhibiting growth of both bacteria. A. hydrophilawas the more sensitive to the two treatments, with 4 log₁₀ cfu g⁻¹less growth occurring at 14 days at 5°C on eugenol-treated breast meat than on control samples. These results suggested that plant extracts might be useful as antimicrobials in cooked, ready-to-eat chicken meat.     

Identification of cultivable lactic acid bacteria isolated from Algerian raw goat's milk and evaluation of their technological properties

One hundred and fifty-eight strains of lactic acid bacteria isolated from Algerian raw goat's milk were identified and technologically characterized. Five genera were found: Lactobacillus (50.63%), Lactococcus (25.94%), Streptococcus (14.56%), Leuconostoc (7.59%) and Pediococcus (1.26%). The predominant species were Lactococcus lactis (32 strains), Streptococcus thermophilus (23 strains), Lactobacillus bulgaricus (19 strains), Lb. helveticus (16 strains) and Lb. plantarum (14 strains).Approximately 39% of the lactic acid bacteria isolated produced more than 0.6% lactic acid (w/v) after 18 h of incubation, and belonged to the Lactococcus and Lactobacillus genera. The highest proteolytic activity was approximately 3 mg tyrosine l⁻¹ for mesophilic strains and nearly 5 mg tyrosine l⁻¹ for thermophilic lactobacilli after 72 h. High aromatic activity (more than 0.8 mg diacetyl l⁻¹ after 16 h) was detected in 14% of the strains.Nine strains were used to make dairy products (a yoghurt-like product and Edam-type cheese) on a pilot scale in the laboratory. The best-liked organoleptic characteristics were noted in a yoghurt produced with a mixed culture made up of S. thermophilus (strain 16TMC+) and Lb. helveticus (strain 20TMC) and in a cheese made with a starter composed of Lc. lactis subsp. lactis (strain 10MCM) and L. lactis subsp. lactis (V.P. +) (strain 19MCM).     

Microbiological,compositional, biochemical and textural characterisation of Caciocavallo Pugliese cheese during ripening

A microbiological, compositional, biochemical and textural characterisation of the pasta filata Caciocavallo Pugliese cheese during ripening is reported. Fully ripened cheese contained a total of ca. log 8.0 cfu g⁻¹ mesophilic bacteria and ca. log 6.0 cfu g⁻¹ presumptive staphylococci, while the number of thermophilic and mesophilic rod and coccus lactic acid bacteria varied during ripening. A two-step RAPD-PCR protocol was used to differentiate biotypes. The natural whey starter was composed mainly of Lactobacillus delbrueckii, Lb. fermentum, Lb. gasseri, Lb. helveticus and Streptococcus thermophilus strains. After day 1 of ripening, Lb. delbrueckii became dominant and some strains of Enterococcus durans and E. faecalis appeared. Non-starter lactic acid bacteria, such as Lb. parabuchneri and Lb. paracasei subsp. paracasei formed a large part of the lactic microflora at 42 and 60 d of ripening. The level of pH 4.6-soluble nitrogen increased from the outer to the inner of the cheese and also increased in each section as ripening progressed, attaining values of 18–15%. Urea-PAGE electrophoresis showed that degradation of α_s1-casein was more rapid than that of β-casein throughout ripening and the rates at which both caseins were degraded greatly increased from the outside to the inside of the cheese. Based on the primary proteolysis products, both chymosin and plasmin appeared to be active. RP-HPLC profiles of the 70% ethanol-soluble, pH 4.6-soluble nitrogen, showed a large number of peaks, indicating a heterogeneous mixture of proteolytic products. There were both age- and section-related changes in the area of the different peptide peaks. Butyric (C4:0), caproic (C6:0), palmitic (C16:0) and oleic (C18:1) acids were the free fatty acids found at the highest concentrations. The level of short chain fatty acids (e.g., butyric and caproic) decreased from the middle and inner to outer sections of the cheese. Peptidase activity in the curd was pronounced, increased during ripening and varied with the cheese section. The greatest increase of the peptidase activity coincided with a change in the lactic microflora and with the prevalence of non-starter lactic acid bacteria. Microbial esterases were supposed to be active together with rennet paste. Little change in the firmness and fractures stress during maturation were found by textural analyses of the raw cheese. The flowability was similar to that of typical low-moisture Mozzarella cheese, while stretchability was lower. The heat-induced changes in phase angle of Caciocavallo Pugliese cheese indicated a phase transition from largely elastic rheological characteristics in unheated cheese to a more viscous and fluid character in melted cheese.     

The influence of multi stage alginate coating on survivability of potential probiotic bacteria in simulated gastric and intestinal juice

The probiotics, Lactobacillus acidophilus PTCC1643 and Lactobacillus rhamnosus PTCC1637, were encapsulated into uncoated calcium alginate beads and the same beads were coated with one or two layers of sodium alginate with the objective of enhancing survival during exposure to the adverse conditions of the gastro-intestinal tract. The survivability of the strains, was expressed as the destructive value (decimal reduction time). Particle size distribution was measured using laser diffraction technique. The thickness of the alginate beads increased with the addition of coating layers. No differences were detectable in the bead appearance by scanning electron microscopy (SEM). The alginate coat prevented acid-induced reduction of the strains in simulated gastric juice (pH 1.5, 2 h), resulting in significantly (P < 0.05) higher numbers of survivors. After incubation in simulated gastric (60 min) and intestinal juices (pH 7.25, 2 h), number of surviving cells were 6.5 log cfu mL^?1 for L. acidophilus and 7.6 log cfu mL^?1 for L. rhamnosus by double layer coated alginate microspheres, respectively, while 2.3 and 2.0 log cfu mL^?1 were obtained for free cells, respectively.