Silicates characterization as potential bacteriocin-carriers

Two different silicates, zeosil and expanded perlite, were characterized as potential carriers of a bacteriocin with anti-Listeria monocytogenes activity, produced by Enterococcus faecium CRL1385. Specific surface areas showed a value significantly higher for zeosil (146 m² g^? 1) than for perlite (0.65 m² g^? 1). Potential zeta measurements revealed that both silicates had negatively charged surfaces between pH 2 and 11, but zeosil presented zero charge near pH 2. Fourier Transform Infrared (FTIR) spectra proved that zeosil presented more silanol groups available for bacteriocin interaction than perlite. Bacteriocins present in the cell-free supernatant (CFS) were adsorbed by both silicates. Adsorption was highest from pH 4 to 8 and, regardless of exposure time (0.5 or 4 h) and silicate concentration (1 or 4% w/v) at 25 °C. Bacteriocin adsorption onto zeosil (ca. 99%) was higher than onto expanded perlite (ca 80%). However, antimicrobial activity of bacteriocins adsorbed onto perlite was higher than onto zeosil. After 2 h contact between L. monocytogenes 01/155 cells and each silicate plus the bacteriocin sample, the number of Listeria viable cells decreased close to 2 and 6 log orders for zeosil and expanded perlite, respectively.Industrial relevance: One of the crucial problems in the use of bacteriocins as food biopreservatives is obtaining and purifying these antimicrobials. The process generally has a poor yield and is industrially expensive. Hence, alternative techniques to deliver bacteriocins may be a likely option to encourage their use as bioprotectors. Silicates, inert compounds of large surface area, are suggested in this work as peptide immobilizers so that they may later be used in food. These inorganic compounds have already been authorized as food-grade anticaking, clarifying or filtering agents. The results achieved so far with adsorption and anti-Listeria activity preservation of bacteriocin, once they have been immobilized onto silicates, offer a promising and simple alternative to incorporate this compound into food.     

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.     

Survival of probiotic bacteria in a whey cheese vector submitted to environmental conditions prevailing in the gastrointestinal tract

Various foods may be used to deliver probiotic bacteria into the gastrointestinal tract; one such example is Requeijão, a Portuguese whey cheese. Survival and stability of Bifidobacterium animalis strains BLC-1, Bb-12, and Bo, Lactobacillus acidophilus strains LAC-1 and Ki, L. paracasei ssp. paracasei strain LCS-1 and L. brevis strain LMG 6906 inoculated into Requeijão, when exposed to simulated gastrointestinal tract conditions, were assessed. Homogenates of inoculated whey cheese in 0.85% (w/v) sterile saline water were exposed to a solution of hydrochloric acid (pH 2.5–3.0) and pepsin (1000 units mL^–1) at 37 °C, and then to 0.3% (w/v) bile salts after 60 or 120 min of acid exposure. All bacterial strains retained their initial viable cell numbers. Bifidobacterium animalis strains Bb-12 and Bo, and L. brevis strain LMG 6906 exhibited the highest viable cell numbers when exposed to bile salts, whereas the other strains had variable death rates.     

Mesophilic lactobacilli in Fiore Sardo cheese: PCR-identification and evolution during cheese ripening

The mesophilic lactobacilli colonizing Fiore Sardo ewe's milk cheese were characterized. They seemed to be the dominant non-starter lactic acid bacteria composing its natural microflora, with a viable cell number varying from 10⁵ CFU g⁻¹ (1-day-old cheese) to 10⁸ CFU g⁻¹ (30-day-old cheese) and then slowly decreasing up to 10⁴ CFU g⁻¹ after 7 months’ ripening. Considering the relevance of mesophilic lactobacilli in affecting the cheese ripening, a PCR-based taxonomic identification of the Lactobacillus species isolated was performed. Cheese samples were collected from 3 farms and 457 isolates from cheeses at different ripening times were analysed with species-specific primers for L. plantarum, L. casei group, L. paracasei, L. casei, L. rhamnosus, L. pentosus, L. paraplantarum, L. curvatus, L. graminis and L. sake. L. plantarum and L. paracasei were the most frequently detected species. Moreover, the development and the evolution during ripening of the facultatively heterofermentative Lactobacillus species (FHL) were different in the three batches of cheese.     

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.     

Fermentative ability of alginate-prebiotic encapsulated Lactobacillus acidophilus and survival under simulated gastrointestinal conditions

Lactobacillus acidophilus was encapsulated in alginate-inulin-xanthan gum and its ability to grow in carrot juice and survive 8 weeks of storage at 4 °C and subsequent exposure to artificial gastrointestinal conditions were assessed. Encapsulation significantly enhanced cell viability after fermentation and storage (6 × 10¹² and 4 × 10¹⁰ cells/ml versus 4 × 10¹⁰ and 2 × 10⁸ for free cells, respectively). Encapsulation protected L. acidophilus from exposure to simulated gastric conditions; minor alterations in viability and the protein profile occurred after incubation in pancreatic juice. For free cells, viability decreased significantly and the expression of numerous proteins was lost after incubation in gastric and pancreatic juice. Thus, encapsulation preserved probiotic bacterial viability and activity; the addition of inulin as a prebiotic component could enhance the functional properties of food products containing this formulation.     

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.     

Influence of probiotic bacteria on the proteolysis profile of a semi-hard cheese

Two probiotic strains, Lactobacillus acidophilus and Lactobacillus paracasei subsp. paracasei, were used as adjunct cultures in semi-hard cheesemaking experiments, in order to study their influence on proteolysis during ripening. Cheeses with and without probiotic bacteria were manufactured. The population of probiotics remained above 10⁷ cfu g⁻¹ during all ripening, and they did not influence primary proteolysis. However, L. acidophilus produced a significant increase in the level of low molecular weight nitrogen compounds and individual free amino acids; the amino acid profiles were also different. Multivariate analysis of peptide profiles showed that samples were grouped mainly by ripening time, although the impact of probiotics was also noticeable. L. acidophilus showed a clear influence on secondary proteolysis, while a minor effect of L. paracasei was evidenced at the end of the ripening. These results showed that the tested strains influenced distinctly proteolysis of cheeses, probably as a consequence of their different proteolytic systems and their activity via the alimentary matrix (cheese).     

Conjugated linoleic acid production by immobilized cells of Lactobacillus delbrueckii ssp. bulgaricus and Lactobacillus acidophilus

Lactobacillus delbrueckii ssp. bulgaricus (CCRC14009) and L. acidophilus (CCRC14079), immobilized with chitosan and polyacrylamide, were tested for CLA production. A 10-ml aliquot of L. delbrueckii ssp. bulgaricus cell suspension (3.59 × 10⁷ CFU/ml) was adsorbed to 0.5 g chitosan and polyacrylamide, mixed with 0.2 ml linoleic acid (0.9 g/ml), and incubated at 37 °C for 24 h at pH 5, 6, 7, and 8 for CLA production. CLA levels, produced by immobilized cells of L. delbrueckii ssp. bulgaricus and L. acidophilus with increasing cell counts to 1.08 and 1.28 × 10¹⁰ CFU/ml, respectively, at optimal reaction pHs were evaluated. More CLA was formed at pH 8 of chitosan and pH 7 of polyacrylamide-immobilized L. delbrueckii ssp. bulgaricus cell treatments. Increase in cell count resulted in higher CLA production. The adsorption of L. delbrueckii ssp. bulgaricus cells onto polyacrylamide at pH 7 showed significant improvement in total CLA level. Results demonstrated a potential for enhancing CLA production through immobilization.     

Production of traditional Greek yoghurt using Lactobacillus strains with probiotic potential as starter adjuncts

Lactobacillus plantarum ACA-DC 146 and L. paracasei subsp. tolerans ACA-DC 4037 were examined for their potential application as adjuncts in the production of traditional Greek set-type yoghurt. Both strains displayed low milk acidification activity, while no inhibition was observed towards or from the yoghurt starters used. Yoghurt produced with L. paracasei subsp. tolerans ACA-DC 4037 exhibited the best sensory properties, with a rich traditional smooth taste, and the strain was selected for further trials. Yoghurt produced with this strain as an adjunct had good physicochemical properties. After 2 weeks of refrigerated storage, microbial loads (>7.0 log cfu g⁻¹) were in accordance with international recommendations and guidelines for probiotic and starter cultures in milk products. Increasing the microbial load further, using concentrated and encapsulated inocula (10–11 log cfu g⁻¹), gave yoghurt with long fermentation times and poor organoleptic properties.     

Histamine accumulation and histamine-forming bacteria in Indian anchovy (Stolephorus indicus)

Accumulation of histamine, trimethylamine (TMA), and total volatile base nitrogen (TVB-N), as well as microbial population incidence in Indian anchovy (Stolephorus indicus) during storage in ice and at 15 and 35 °C were investigated. Histamine was as low as 1.9 mg/100 g in 15 days at ice storage, but it increased to 19.0 mg/100 g after 32 h at 15 °C. Histamine rapidly increased to 25.4 mg/100 g when stored at 35 °C for 8 h. TVB-N and TMA began to sharply increase after 11 days in ice storage, but abruptly increased after 16 and 8 h of storage at 15 and 35 °C, respectively. A high number of Enterobacteriaceae (10¹⁰–10¹¹ cfu/g) was detected and shown to be the dominant group of microbial flora during spoilage of Indian anchovy at both 15 and 35 °C. A total of 153 bacterial strains were selected from the prescreening step using various selective media. Only 75.8% of these selected isolates showed a positive reaction in Niven's differential medium, and 27.6% of the positive isolates were true histamine formers when confirmed by the enzymatic method. Prolific histamine formers were identified as Morganella morganii, Proteus vulgaris, and Enterobacter aerogenes, and produced high histamine content of 104.1–203.0 mg/100 ml. Optimum growth and histamine production of selected strains of these three species was at 35 °C in histamine evaluation broth (HEB) containing 0.5% NaCl, pH 5. E. aerogenes produced the highest histamine of 500 mg/100 ml at the optimum condition. All studied strains did not produce histamine at ⩾10% NaCl.     

Kefir yeasts enhance probiotic potentials of Lactobacillus paracasei H9: The positive effects of coaggregation between the two strains

This study investigated pure Lactobacillus paracasei H9 tolerance to simulated gastrointestinal juices and adhesion to intestinal mucosa cells without yeasts, with viable yeasts (VY) and with different pretreated yeasts. Three models including gastric secretion tolerance (GST), intestinal juice tolerance (IJT) and sequential gastrointestinal tolerance (SGT) were respectively employed to assay the tolerance of L. paracasei H9, whilst Caco-2 cell line was used to investigate the bacterial adhesion. Particularly, the co-aggregation ability of the two strains at pH values of 2.0, 8.0 and 7.2 was originally carried out to study relations to the bacterial probiotic potentials. Results showed that yeast counts in the range from 3.0 to 5.0 log CFU mL^? 1 could gradually increase the viability of L. paracasei H9 in SGT. The bacterial viability in the three tolerance models and the adherent number to Caco-2 cells were significantly improved with addition of VY (P < 0.05). The L. paracasei H9 with VY in gastric juice at pH 2.0 and intestinal juice at pH 8.0, respectively, exhibited higher aggregation percentage compared with that of single L. paracasei H9 at 37 °C (P < 0.05). The aggregation ability of L. paracasei H9 with VY at pH 7.2, which might contribute to increase the adhesion of the bacteria, also excelled that of L. paracasei H9 (P < 0.05). It is deduced that proteins of the bacterial cell surface and polysaccharides in yeast cell walls play important roles in co-aggregation of the two strains and the microbial adhesion specificity to Caco-2 cells. The co-aggregation of the two strains also contributes to enhancing probiotic potentials of L. paracasei H9.     

The efficacy of sand-immobilized organoclays as filtration bed materials for bacteria

Previous research has shown that montmorillonite clays exchanged with cationic surfactants including cetylpyridinium (CP), hexadecyltrimethylammonium (HDTMA), and cetyldimethylethylammonium (CDEA) were effective in reducing Salmonella enteritidis colony counts in bulk antibacterial assays. In order to increase the porosity (and hydraulic conductivity) of these materials for use in filtration beds, organoclays were tightly adhered and immobilized onto the surface of sand. Bulk antibacterial assays performed with 10 mg of the sand-immobilized organoclay (S/IOC) showed that the composite materials retained their antibacterial activity after processing. Reductions in plate colony counts ranged from 16.5% for sand-immobilized CP-exchanged sodium montmorillonite to 83.9% for immobilized CDEA-exchanged low pH montmorillonite. The sand/adhesive matrix and sand-immobilized activated charcoal failed to produce any significant decreases. Pilot studies with columns containing 1.0 g of S/IOC showed that maximal filtration efficiency was obtained at a filtration rate of 1.0 ml/min. Initial column studies with Salmonella suspensions at a concentration of 3.0×10⁸ cfu/ml, produced reductions in colony plate counts varying from 28.5% for immobilized CP-exchanged calcium montmorillonite to 59.5% for immobilized CDEA-exchanged low pH montmorillonite. The sand/adhesive matrix and immobilized activated charcoal yielded much lower reductions (7.6% and 10.4%, respectively). Studies with Escherichia coli suspensions (3.0×10⁷ cfu/ml) indicated reductions varying from 59.3% for immobilized CP-exchanged calcium montmorillonite to 99.9% for immobilized CDEA-exchanged low pH montmorillonite. These initial results suggest that S/IOC can be effective in reducing microbial numbers in wastewater following some modifications.     

The protective effect of monosodium glutamate on survival of Lactobacillus rhamnosus GG and Lactobacillus rhamnosus E-97800 (E800) strains during spray-drying and storage in trehalose-containing powders

The use of trehalose as a means of preserving Lactobacillus rhamnosus GG (LGG) and L. rhamnosus E-97800 (E800) during spray-drying and the effects of incorporated monosodium glutamate (MSG) in the carrier medium on the survival rates during drying and storage were examined. E800 was more resistant to heat than LGG in 20%, w/w, trehalose; the d-values at 65 °C were 14 s and 5.1 s, respectively. An air outlet temperature of 65–70 °C was taken as optimal for the drying process, as the resultant moisture levels in trehalose containing these bacteria were 4.1% (w/w) and 3.79% (w/w) with corresponding viable counts of 3.65 × 10⁸ cfu mL^?1 and 1.80 × 10⁹ cfu mL^?1, respectively. The presence of MSG increased the final viable counts of LGG and E800 to 3.05 × 10⁹ cfu mL^?1 and 1.30 × 10⁹ cfu mL^?1, respectively. Survival of LGG and E800 remained constant at a minimum level of ～10⁸ cfu mL^?1 during storage at 25 °C in trehalose–MSG medium.     

Comparative Studies of Ginger (Zingiber officinale) and West African Black Pepper (Piper guineense) Extracts at Different Concentrations on the Microbial Quality of Soymilk and Kunun-zaki

Extracts of two spices, namely ginger (Zingiber officinale) and black pepper (Piper guineense) were prepared in 0.4%, 1.2%, 2.4% and 3.6% concentrations. Soymilk and kunun-zaki were treated respectively with the different concentrations and stored at ambient temperature for 5 days. The microbial load and identification were determined every day of storage until samples were adjudged spoilt.On the first day, 0.4% ginger extract in soymilk and kunun-zaki had a microbial load of 7.77 × 10^6b and 5.17 × 10^6b respectively. 3.6% ginger extract in soymilk and kunun-zaki recorded 3.73 × 10^6b and 3.30 × 106 each. 0.4% black pepper extract in soymilk had 6.273 × 10^6b and recorded 4.63 × 10^6b in kunun-zaki. 3.6% black pepper extract in soymilk and kunun-zaki had a microbial load of 3.20 × 10^6d and 2.90 × 10^6c respectively. On the third day the microbial load increased for both ginger and black pepper extract. Ginger extract recorded 9.13 × 10^6b in soymilk and 5.60 × 10^6b in kunun-zaki at 0.4% concentration. Black pepper extracts recorded 7.43 × 10^6b in soymilk and 3.27 × 10^6b in kunun-zaki also at 0.4% extract. 3.6% black pepper extract recorded 4.10 × 10^6a in soymilk and 2.20 × 106c in kunun-zaki.There was linear reduction of microbial load as spice concentration increased. Black pepper recorded lower microbial load, thus having more antimicrobial activity and may be preferred to be used as natural antimicrobial preservatives to extend the shelf-life of food.     

Combinations of nisin with organic acids or salts to control Listeria monocytogenes on sliced pork bologna stored at 4°C in vacuum packages

This study evaluated dipping solutions of nisin with or without organic acids or salts, as inhibitors of Listeria monocytogenes introduced on sliced cooked pork bologna before vacuum packaging and storage at 4°C for 120 days. Inoculated (10²–10³ cfu/cm²) slices were immersed in nisin (5000 IU/ml), or in lactic or acetic acid (1, 3, 5 g/100 ml), sodium acetate or diacetate (3, 5 g/100 ml), and potassium benzoate or sorbate (3 g/100 ml), each combined with nisin. Additional slices were immersed in nisin, inoculated and then immersed in acid or salt solutions without nisin. Nisin reduced L. monocytogenes by 1.0–1.5 log cfu/cm² at treatment (day-0) followed by a listeriostatic effect for 10 days. Thereafter, however, the pathogen multiplied in treatments without acid or salts, with growth being faster on slices immersed in nisin after as compared to before inoculation. Nisin in combination with 3 or 5 g/100 ml acetic acid or sodium diacetate or 3 g/100 ml potassium benzoate, applied individually or as mixtures, did not permit growth before day-90. Other treatments were of variable and lesser effectiveness (20–70 days), whereas in untreated or water-treated (control) bologna L. monocytogenes increased at 6–7 log cfu/cm² at day-20. Based on the antilisterial efficacy and effects of treatments on product pH, nisin with 3 g/100 ml sodium diacetate may be the most promising combination in dipping solutions to control L. monocytogenes on sliced cured pork bologna.     

Antimicrobial plastic film: Physico-chemical characterization and nisin desorption modeling

Antimicrobial active films represent an innovative concept in food packaging, developed to answer to consumer's expectation for better microbiological safety. In this study, the growth of pathogenic micro-organisms on the surface of food is proposed to be controlled by coating, on the surface of polyethylene/polyamide/polyethylene film (PE/PA/PE), a film-forming solution containing Nisaplin, a commercial form of bacteriocin produced by Lactococcus lactis subsp. lactis: nisin. The bioactivity of these multi-layer films coated with Nisaplin loaded HydroxyPropylMethylCellulose film is based on the release of this antimicrobial molecule towards a food simulant. Nisin mass transfer was studied and modeled, for different operating conditions, generally encountered in food products. pH didn't seem to interfere with nisin release kinetics, while the variation of NaCl concentration between 0.8% and 3.2% decreased the desorption coefficient (k_d) by 18% and the temperature increase from 10 °C to 28 °C resulted in an increase of k_d from 1.78 × 10^? 2 m s^? 1 to 2.10 × 10^? 2 m s^? 1. Coating of PE/PA/PE film with this antimicrobial layer induced little mechanical properties modifications without compromising industrial applications. Water barrier capacity was not altered.Industrial relevanceThis paper concerns active packaging, considered as a new approach to preserve food shelf life. Active packaging is a real gain for plastic and Food industrials. Coating was used to obtain antimicrobial packaging. The impact of coating on film characteristics is investigated.Also, antimicrobial agent desorption is determined during storage conditions.     

Assessing the acid tolerance and the technological robustness of probiotic cultures for fortification in fruit juices

The suitability of probiotic cultures as fruit juice supplements was examined by assessing their acid tolerance and technological robustness. Survival of Lactobacillus and Bifidobacterium strains in orange juice (OJ), pineapple juice (PJ) and cranberry juice (CJ) was monitored. Results revealed that extensive differences exist among probiotic strains regarding their acid resistance. All of the strains screened survived for longer in OJ and PJ compared to CJ. L. casei DN-114 001, L. rhamnosus GG and L. paracasei NFBC43338 displayed the greatest robustness surviving at levels above 10⁷ cfu ml^− 1 in OJ and above 10⁶ cfu ml^− 1 in PJ for at least 12 weeks. Probiotic tolerance to thermal and non-thermal processing was studied to determine the feasibility of their addition to OJ prior to pasteurisation. OJ fortified with probiotic cultures was subjected thermal pasteurisation at 76 °C for 30 s and 90 °C for 1 min in addition to a high pressure treatment of 400 MPa for 5 min. Results indicated no strain was capable of withstanding treatments necessary to achieve a stable juice at levels > 10⁶ cfu ml^− 1. The outcome of the overall study points to L. rhamnosus GG, L. casei DN-114 001 and L. paracasei NFBC43338 as having promising potential for exploitation as functional supplements in fruit juices due to their impressive tolerance in acidic environments; however, fortification post processing is recommended.Industrial relevanceThe ability of health-promoting cultures to survive for at least 12 weeks in orange juice and pineapple juice at commercially critical levels renders them suitable strains for exploitation. Their inclusion may enhance the market potential of these already successful beverages.     

Effect of different activated coatings containing enterocin AS-48 against Listeria monocytogenes on apple cubes

Enterocin AS-48 is a circular bacteriocin with strong anti-Listeria activity. The purpose of the present study was to evaluate the effect of the bacteriocin incorporated into different coating solutions on a cocktail of five L. monocytogenes strains previously inoculated on apple cubes. Coating solutions were made with chitosan, caseinate, alginate, k-carrageenate, xanthan gum, pectin, starch, carboxymethyl cellulose or methyl cellulose. Coatings were applied singly or combined with enterocin AS-48 at 20 or 40 μg/ml. Samples were stored at 4 °C for 7 days. The single application of coatings had almost no effect (as in alginate and methyl cellulose) or had a low effect on Listeria viability (< 2.0 log cycles), with the exception of chitosan coating which showed a strong anti-Listeria activity (up to 3.7 log cycles at day 7). Coatings dosed with 20-μg/ml enterocin AS-48 reduced viable Listeria counts gradually during storage in most cases, achieving significant reductions (p < 0.05) of 1.0 to 1.9 log cycles after 7 days for k-carrageenate, xanthan gum, pectin, starch, carboxymethyl cellulose and methyl cellulose compared to the single coating. At 40 μg/ml, enterocin AS-48 significantly reduced viable counts (p < 0.05) for most coatings (by 1.4 to 3.3 log cycles, depending on the coating) compared with coatings without bacteriocin (except for chitosan). Chitosan, pectin, xanthan gum and carboxymethyl cellulose coatings, supplemented or not with 40 μg/ml AS-48 were further investigated in combination with 20 mM EDTA or with 2.0% sodium lactate. The single addition of sodium lactate showed the greatest effects at day 7, where it reduced viable counts significantly (p < 0.05) by 1.1 to 2.2 log cycles compared to the single coatings (except for chitosan), whereas the combination of sodium lactate and AS-48 reduced viable counts below detection levels also at day 7 for all coatings. The combination of EDTA and AS-48 was much more effective, reducing Listeria counts below detection levels from day 1 for most of the coatings tested. The combination of EDTA and AS-48 was also the most effective at time 0, achieving reductions of viable counts between 2.0 and 2.7 log cycles depending on the coating immediately after treatment compared with single coatings.Industrial relevanceResults from the present study suggest the potential of edible coatings containing enterocin AS-48 and EDTA for inactivation of L. monocytogenes on apple surfaces. Since edible coatings are widely used on fruit surfaces, coatings activated with enterocin AS-48 and EDTA could find application as a hurdle against L. monocytogenes in fresh-cut apple pieces.     

Milk Somatic Cells and Lactation in Small Ruminants

The milk somatic cell count (MSCC) is the basis for abnormal milk control programs. The current legal MSCC limit for bulk tank milk for goats and sheep in the United States is 1000 and 750 × 10³/ml, respectively. Milk somatic cell counts for goats are higher than MSCC for cows and sheep. The MSCC for goats free from intramammary infection (IMI) range from 270 to 2,000 × 10³/ml. Cell counts for sheep are similar to cows and range from 10 to 200 × 10³/ml. Neutro-phils comprise the major cell type in milk from uninfected goats and constitute 45 to 74% of the MSCC, compared with 2 to 28% for sheep and cows. The macrophage is the major cell type in milk from cows and sheep. Milk secretion in goats and sheep is largely apocrine in nature and cytoplasmic particles, similar in size to milk somatic cells, are normal constituents of their milk. Concentrations of cytoplasmic particles in sheep milk average 15 × 10³/ml, while goat milk averages 150 × 10³/ml. Therefore, to obtain accurate MSCC for goats, only cell counting procedures specific for DNA should be used. While IMI significantly increases MSCC for goats and sheep, noninfectious factors such as parity, stage of lactation, season and milk yield have been related to increased MSCC. An increase in MSCC for goats has been shown to decrease milk and fat yields. Intramammary infusion of antibiotics at dry-off and postmilking teat dipping in goats decreased the rate of new IMI and MSCC. Thus, mastitis control practices shown to be efficacious in cows are also effective in goats.