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.     

Effect of pre-incubation conditions on growth and survival of Staphylococcus aureus in sliced cooked chicken breast

In this work, the effect of pre-incubation conditions (temperature: 10, 15, 37 °C; pH 5.5, 6.5 and water activity, a_w: 0.997, 0.960) was evaluated on the subsequent growth, survival and enterotoxin production (SE) of Staphylococcus aureus in cooked chicken breast incubated at 10 and 20 °C. Results showed the ability of S. aureus to survive at 10 °C when pre-incubated at low a_w (0.960) what could constitute a food risk if osmotic stressed cells of S. aureus which form biofilms survive on dried surfaces, and they are transferred to cooked meat products by cross-contamination. Regarding growth at 20 °C, cells pre-incubated at pH 5.5 and a_w 0.960 had a longer lag phase and a slower maximum growth rate. On the contrary, it was highlighted that pre-incubation at optimal conditions (37 °C/pH 6.5/a_w 0.997) produced a better adaptation and a faster growth in meat products what would lead to a higher SE production. These findings can support the adoption of management strategies and preventive measures in food industries leading to avoid growth and SE production in meat products.     

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.     

Biotyping and enterotoxigenicity of Staphylococci isolated from fresh and frozen meat marketed in Jordan

A total of 286 fresh and processed meat samples marketed in Jordan were collected for isolation and typing of Staphylococci. Devriese’s system was followed for biotyping of the isolated Staphylococcus aureus subsp. aureus and enterotoxigenicity of the isolates was also determined. S. aureus subsp. aureus was found in 80.8% of the samples. Means counts for meats of the various sources ranged from 5.3 × 10² to 4.3 × 10⁴ cfu/g. However, only 33.6% of the samples had coagulase positive Staphylococci. Biotyping of S. aureus subsp. aureus isolates revealed that 12%, 24%, 8%, and 27% are human, bovine, ovine and non-host specific biovars, respectively. Only 28% of these isolates, of unknown type, were isolated from goat’s meat. Enterotoxigenic S. aureus subsp. aureus isolates accounted for 13.5% of a total of 231 Staphylococci. The non-S. aureus enterotoxigenic isolates were identified as Staphylococcus epidermidis, Staphylococcus sciuri and Staphylococcus intermedius. Staphylococci enterotoxin types A, C, and B accounted for 57%, 36% and 7% of the isolated S. aureus subsp. aureus, respectively. This study showed that the enterotoxigenic Staphylococci represent a potential hazard in meat and the Devriese’s biotyping system needs to be extended to accommodate the isolates of unknown type.     

Effects of drying methods and conditions on antimicrobial activity of edible chitosan films enriched with galangal extract

The aim of this work was to study the effects of drying methods and conditions (i.e., ambient drying, hot air drying at 40 °C, vacuum drying and low-pressure superheated steam drying within the temperature range of 70–90 °C at an absolute pressure of 10 kPa) as well as the concentration of galangal extract on the antimicrobial activity of edible chitosan films against Staphylococcus aureus. Galangal extract was added to the film forming solution as a natural antimicrobial agent in the concentration range of 0.3–0.9 g/100 g. Fourier transform infrared (FTIR) spectra and swelling of the films were also evaluated to investigate interaction between chitosan and the galangal extract. The antimicrobial activity of the films was evaluated by the disc diffusion and viable cell count method, while the morphology of bacteria treated with the antimicrobial films was observed via transmission electron microscopy (TEM). The antimicrobial activity, swelling and functional group interaction of the antimicrobial films were found to be affected by the drying methods and conditions as well as the concentration of the galangal extract. The electron microscopic observations revealed that cell wall and cell membrane of S. aureus treated by the antimicrobial films were significantly damaged.     

Microbiological and chemical changes during the manufacture of Kefir made from cows’ milk,using a commercial starter culture

The counts of total aerobic mesophilic bacteria, lactic acid bacteria (in three different culture media, M17 agar, MSE agar, and Rogosa agar) and yeasts, and some biochemical parameters (levels of lactose, glucose, galactose, L(+)- and D(−)-lactic acids, ethanol, titratable acidity and pH) were determined during 196 h of fermentation in five batches of Kefir made from cows’ milk using a commercial starter culture. Lactococcus spp. predominated during the first 48 h of fermentation (∼8 log₁₀ cfu g⁻¹); Lactobacillus spp. became the predominant species after 48 h (∼8.5 log₁₀ cfu g⁻¹). During the first 24 h of fermentation, the lactose content decreased from a mean value of 4.92% (w/w) to 4.02% (w/w); the concentration of L(+)-lactic acid increased from 0.01% to 0.76% (w/w) and the pH decreased to 4.24 over the same period. After 24 h of fermentation, the changes in the levels of lactose and L(+)-lactic acid, and in pH, occurred more slowly. Neither glucose nor galactose were detected during fermentation. The production of ethanol was limited, reaching a mean final value of 0.018% (w/w).     

Viscosity of model yogurt systems enriched with barley β-glucan as influenced by starter cultures

The effect of barley β-glucan (BG) on the growth of two yogurt starter cultures (SCs) each consisting of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was investigated. Model yogurt systems were prepared with or without the incorporation of barley BG and lactose to mimic natural levels in milk and allowed to ferment for 8 h at 42 °C. The fermentation efficacy of the SCs was monitored through pH measurements. Samples were collected and analyzed for pH and viscosity at 0, 2, 4, and 8 h. Viscosity was determined using a rotational viscometer at shear rates of 1.29–129 s⁻¹ and 20 °C. The pH measurements indicated no adverse effect on the growth of SCs as the acidification process was consistent with normal pH development, but viscosity measurements suggested that BG was depolymerized by SCs when lactose became a limiting nutrient during fermentation.     

X-ray diffraction analysis of lactose crystallization in freeze-dried lactose–whey protein systems

Water sorption, time-dependent crystallization and XRD patterns of lactose and lactose–WPI mixtures were studied with glass transition data. The results indicated that the sorbed water of lactose–WPI mixtures was fractional and water content of individual amorphous components in lactose–WPI mixtures at each a_w from 25 °C to 45 °C could be calculated. Crystallization occurred in pure lactose whereas partial crystallization was typical of lactose–WPI mixtures (protein content ≤ 50%) at intermediate and high a_w (> 0.44 a_w) from 25 °C to 45 °C. The extents of crystallization were significantly delayed by WPI. The T_g values of lactose–WPI systems showed the composition-dependent property in systems and might indicate the occurrence of phase separation phenomena during 240 h storage. XRD showed no anhydrous β-lactose and mixed α-/β-lactose with molar ratios of 4:1 crystals in crystallized lactose–WPI systems (70:30 and 50:50 solids ratios). Reduced crystallization in the presence of WPI was more pronounced possibly because of reduced nucleation and diffusion during crystal-growth. The present study showed that WPI could present an important role in preventing sugar crystallization.     

Continuous Production of Lactic Acid from Whey Perméate by Free and Calcium Alginate Entrapped Lactobacillus helveticus

Lactobacillus helveticus strain milano was used for the continuous fermentation of lactic acid in cheese whey-yeast extract permeate medium. The best productivity of lactic acid was with the free cell system, which was 9.7 g/L per h at a dilution rate of .352 h⁻¹. Under such conditions, lactose conversion was 87.5%, based on the lactose concentration of 37.4 g/L in feed. Under high dilution rates, the cells were elongated to several times their normal size, resulting in wall growth. The cell growth on the fermentor wall caused system instability; however, it prevented cell wash-out under high dilution rates. The packed bed column system using Ca alginate entrapped cells is not suitable for practical application. Nonuniform pH control, plugging of the column by leaked cells, and decalcification of Ca alginate beads were major drawbacks of the packed bed system.     

Preliminary characterization of bacteriocins from Lactococcus lactis,Enterococcus faecium and Enterococcus mundtii strains isolated from turbot (Psetta maxima)

The aim of this work was the characterization of new strains of lactic acid bacteria (LAB) from farmed fish and with potential application as biopreservatives against both Listeria monocytogenes and Staphylococcus aureus. Twenty-five strains of LAB isolated from the muscle of farmed turbot were investigated. Genetic identification of the bacteriocin-producing LAB strains was performed by means of a PCR method using novel BAL1/BAL2 16S ribosomal-RNA-targeted primers. Maximum bacteriocin production by Lactococcus lactis ssp. lactis USC-39, Enterococcus faecium USC-46 and Enterococcus mundtii USC-51 was detected in the stationary phase of growth. Both acidification and the production of hydrogen peroxide by LAB were ruled out as the source of the inhibition. In contrast, the antimicrobial activity of all three LAB strains was inactivated by the addition of proteinase K, thus confirming the proteinaceous nature of the inhibition. The activity against L. monocytogenes was maintained in the 3.5–5.5 or 3.5–6.5 pH range, depending on the LAB strain. Likewise, inhibition of S. aureus strains was observed in the 3.5–4.5 and in the 3.5–5.5 pH ranges, depending on the LAB strain and on the S. aureus strain tested. Bacteriocin activity was stable in all three strains after heating the cell-free extract for 60 min at 100 °C, or even for 15 min at 121 °C, in all the three LAB strains. The acidic and heat-resistant bacteriocins produced by the three LAB strains isolated from turbot, able to inhibit the growth of both L. monocytogenes and S. aureus may find application as biopreservatives in fermented and/or heated food products.     

Enzymatic Production of Ethanol from Cassava Starch Using Two Strains of Saccharomyces cerevisiae

Cassava starch from TMS 30572 and Idileru were hydrolyzed with α-amylase and amylo-glucosidase before fermentation using two strains of Saccharomyces cerevisiae from palm wine and bakers’ yeast. The per cent yield of sugars and total dissolved solids were 66 % and 26% respectively while pH was 7. Spectrophotometric measurement of the cell growth revealed steady but insignificant (p ≤ 0.05) increase in cell concentrations up to 48 h fermentation time with a gradual decline by 72 h. Saccharomyces cerevisiae strain from palm wine grew best on TMS 30572 hydrolysate at 20% sugar concentration (optical density 0.663; fermentation time 48 h) while on Idileru hydrolysate it grew best at 25% (optical density 0.698; fermentation time 60 h). The pH values obtained from the fermenting hydrolysates for both yeast strains declined gradually as the fermentation progressed with the lowest pH values (3.01 for S. cerevisiae from palm wine; 3.06 for S. cerevisiae from bakers’ yeast) obtained for TMS 30572 cassava variety at 25% sugar concentration. Changes in pH were significant (p ≤ 0.05). The Saccharomyces cerevisiae strain from palm-wine had a higher conversion of available sugar into ethanol. The yield of ethanol was found to vary but the highest ethanol concentration obtained was 5.3% at 10% initial sugar concentration, which gave a sugar conversion efficiency of 37.3%. The results obtained suggest that Saccharomyces cerevisiae strains from sources other than those used conventionally can serve as good substitutes for bio-conversion processes in the industrial production of ethanol.     

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.     

Biocontrol of Staphylococcus aureus in curd manufacturing processes using bacteriophages

The ability of specific bacteriophages to inhibit Staphylococcus aureus growth in curd manufacturing processes was determined. Two lytic bacteriophages specific against S. aureus were obtained by DNA random deletion from the milk-isolated temperate phages, ΦH5 and ΦA72. A cocktail of these lytic phages, Φ88 and Φ35, at multiplicity of infection (MOI) of 100, produced a complete elimination of 3×10⁶ cfu mL⁻¹ of the pathogen in ultra-high-temperature (UHT) whole milk at 37 °C. Furthermore, the frequency of emergence of bacteriophage-insensitive mutants was reduced up to 200-fold in the presence of the two lytic phages compared with that detected with the combination of the temperate counterparts. The lytic phage derivatives, added to milk, were able to decrease rapidly the viable counts of S. aureus during curd manufacture. In acid curd, the pathogen was not detected after 4 h of incubation at 25 °C, whereas pathogen clearance was achieved within 1 h of incubation at 30 °C in renneted curd. These results indicate that lytic bacteriophages could be used as biopreservatives in the manufacture of particular dairy products.     

Volatile components and antibacterial effects of pine needle (Pinus densiflora S. and Z.) extracts

The antibacterial effects of the volatile components extracted from Pinus densiflora S. and Z., by simultaneous steam distillation and solvent extraction (SDE), were examined on six foodborne bacteria using Bioscreen C (a computer-controlled shake–incubator–reader). The SDE extracts of P. densiflora obtained after 1.5 or 2.0 h at pH 3.6 exhibited a strong growth inhibitory effect on Escherichia coli O157:H7 and their overall antibacterial activities against the various bacteria tested tended to increase with increased extraction time and with a lower extraction pH. The major volatile components of the SDE extracts obtained at pH 3.6 and 1.5 h, as determined by gas chromatography, were α-ocimene (29.3%), sabinene (10.9%), β-myrcene (9.6%), β-caryophyllene (8.0%), β-cadinene (7.3%), α-terpinolene (4.9%), 2-hexanal (4.5%), and β-pinene (4.3%). The addition of 8% or 10% (v/v) of the SDE extracts to culture broth completely inhibited the growths of Bacillus cereus, Salmonella Typhimurium, and Staphylococcus aureus. The intracellular adenosine triphosphate (ATP) concentration of S. Typhimurium treated with P. densiflora extract reduced to 0.165 μm from 0.595 μm, whereas the ATP concentration in culture supernatants was increased to 0.469 μm form 0.065 μm.     

Lactose isomerization into lactulose in an electro-activation reactor and high-performance liquid chromatography (HPLC) monitoring of the process

Lactulose production is a complex process that is affected by several operational and experimental conditions. This study investigated the production of prebiotic lactulose using an electro-activation reactor. The experiments were performed at an ambient temperature (22 ± 1 °C), and the lactose starting concentration was constantly maintained at 5% (w/v). This work used HPLC to investigate the retention behaviors of the disaccharides (lactulose and lactose) and the by-products of lactulose production (glucose, galactose and fructose) under varying experimental conditions such as electric potential difference, central cell compartment electrolyte concentration and electro-activation time. The results were then correlated with the electric current density, the electrical global resistance of the system, the pH, the temperature and the Joule heating effect. The highest lactulose yield that was obtained during the isomerization of lactose was ≈30% at a voltage of U = 28 V, an electrolyte concentration C_Na2SO4 = 0.5 M, a temperature T = 26 °C, an alkaline medium pH = 10.79, and a running time τ = 20 min.     

Antibiotic resistance and susceptibility to some food preservative measures of spoilage and pathogenic micro-organisms from spices

Antibiogram of 84 strains of Bacillus cereus, 26 strains of Clostridium perfringens, four strains of Staphylococcus aureus, 51 strains of Enterobacteriaceae, two strains of each of Salmonella and Shigella; isolated from spices, were studied against 20 different antibiotics that are commonly used against foodborne diseases, mainly gastroenteritis. All the tested strains of B. cereus, Cl. perfringens, Staph. aureus, Escherichia coli, Salmonella and Shigella were found resistant to at least 3, 4, 7, 6, 10 and 9 antibiotics, respectively. In brain–heart infusion broth supplemented with glucose, the D_100°C-values for B. cereus were 3.5–5.9 min, and the z-values were 17–18°C. The D_100°C-values for Cl. perfringens in fluid thioglycolate medium were higher (10.0–19.8 min) than those of B. cereus. The minimum inhibitory concentrations (MICs) of sodium chloride were 45–80 mg ml⁻¹. While the MIC of benzoic acid for Cl. perfringens, tested on perfringens agar (pH 7.3) plates by incubating anaerobially at 35°C for 24 h, was 1.9–2.2 mg ml⁻¹, for others, tested on nutrient agar (pH 6.8) plates by incubating at 35°C for 18 h in static aerobic condition, it was much less. Similarly, the MIC of sorbic acid for all the tested isolates, excepting Cl. perfringens, was 0.6–1.1 mg ml⁻¹. Of the eight isolates of Cl. perfringens, only three were inhibited at 2.0 mg sorbic acid ml⁻¹, while others were resistant. Sixty percent and 75% of the respective strains of B. cereus and Cl. perfringens were resistant to 5000 IU Nisaplin ml⁻¹, whereas the MIC values of Staph. aureus were between 3000 and 5000 IU ml⁻¹. While studying combined effect of selected hurdles on the growth of enterotoxigenic Cl. perfringens 16-C2, the judicious combination considered was low acid (pH 6.0), 30 mg sodium chloride ml⁻¹ and 1.25 mg benzoic acid ml⁻¹.     

Effects of high pressure treatment on glycolytic enzymes of Lactococcus lactis subsp. lactis,Streptococcus thermophilus and Lactobacillus acidophilus

High pressure processing (HPP) reduces the glycolytic activity of lactic acid bacteria (LAB) and provides a means to control further production of acidic metabolites in fermented dairy products during storage. However, there is limited information on the effects of HPP on specific enzymes of dairy starter bacteria responsible for the metabolism of lactose. The aim of this study was to determine pressure-induced inactivation of glycolytic enzymes in Lactococcus lactis subsp. lactis C10, Streptococcus thermophilus TS1 and Lactobacillus acidophilus 2400. Cultures were grown for 16 h in M17 or MRS broth containing 5% (w/v) lactose at pH 6.5 (maintained by addition of 10 M NaOH). The cells were harvested by centrifugation, washed and resuspended in 100 mM phosphate buffer (pH 6.5) and pressure-treated at 300 and 600 MPa (≤ 22 °C, 5 min). The ability of pressure-treated resting cells of Lactococcus, incubated with 5% (w/v) lactose at 30 °C, to ferment lactose was evaluated by determining titratable acidity (TA) during incubation. The activities of phospho-β-galactosidase (P-β-gal), β-galactosidase (β-gal) and lactate dehydrogenase (LDH) were determined in cell-free extracts of untreated and pressure-treated cells. Resting cells of Lactococcus treated at 600 MPa had a substantially lower rate of acidification than the controls and those treated at 300 MPa. Both P-β-gal and β-gal were significantly inactivated (p < 0.01) in the starter cultures treated at 300 or 600 MPa. The LDH in Lactococcus and Lactobacillus was highly resistant to pressure treatment at 300 MPa. In contrast, the LDH in Streptococcus was almost completely inactivated at ≥ 300 MPa.Industrial relevanceContinuing production of acidic metabolites in fermented dairy products during storage can be a technological challenge that adversely affects product quality. The current study demonstrates that high pressure processing (HPP) offers the potential of controlling this problem by inactivation of glycolytic enzymes in various mesophilic and thermophilic starter cultures. The findings of this research will assist in establishing optimised operating parameters for HPP treatment of cultured products to extend shelf-life, by reducing acid production during storage.     

The distribution of Staphylococcus sp. on bovine meat from abattoir deboning rooms

In developing countries such as South Africa, Staphylococcus aureus has been shown consistently to be one of the most important micro-organisms responsible for food poisoning outbreaks. In this study, the staphylococci in selected South African abattoirs were quantified, identified and further characterized in terms of coagulase types. The highest staphylococci counts (1.7×10⁶ cfu g⁻¹) were observed in the meat from the high throughput (Grade A) abattoir during week 3. The counts exceeded the National Guidelines (10² cfu g⁻¹) without exception and at least 50% surpassed the levels sufficient to produce toxins (10⁵ cfu g⁻¹) determined for S. aureus. Species were dominated by S. capitis, S. xylosus, S. auricularis, S. aureus and S. intermedius. In terms of the coagulase types of S. aureus, type V was the most dominant and type VI the least. It became evident that the hygiene practices implemented by the abattoirs investigated in this study were not effective enough in reducing the contamination levels of the staphylococci from carcasses. It is therefore recommended that the sampled abattoirs revise their manufacturing strategies in order to reduce the levels of staphylococcal contamination which have been shown to be transferred through food handlers, surfaces, equipment and the environment.     

Influence of pH and NaCl on monolaurin inactivation of Streptococcus iniae

Streptococcus iniae, a Gram-positive bacterium, has been recognized as a cause of human and fish streptococcosis. Aquacultured fish are susceptible to infection and have been epidemiologically linked with wound infections of fish handlers. Therefore, there is a need to develop methods of prevention and/or control of this pathogen. This study determined the minimum inhibitory concentration of monolaurin (glycerol monolaurate) against S. iniae strains PW and BU and utilized gradient plates at 37°C to determine the combined influence of pH and NaCl on monolaurin inactivation of S. iniae. The minimum inhibitory concentration of monolaurin against strain PW was 12.4±0.3 μg ml⁻¹ and against strain BU was 12.1±0 μg ml⁻¹. Compared to growth in the absence of monolaurin, the incorporation of 6 μg ml⁻¹ monolaurin into the salt (1.12–6.95%)–pH (4.26–8.88) gradient plates prevented growth of strain PW at pH values <6.00 and strain BU at pH values <6.20, regardless of the salt concentration. The combination of monolaurin with environmental variables such as pH and salt proved to be an effective tool for in vitro control of S. iniae.     

Quantitative Changes in Oligosaccharides During Fermentation and Storage of Yogurt Inoculated Simultaneously with Starter Culture and β-Galactosidase Preparation

Low lactose yogurt was manufactured by simultaneous inoculation with β-galactosidase of Aspergillus oryzae from two commercial sources and yogurt starter culture. Addition of 4.7 units enzyme/100 ml resulted in 64 and 58% depletion of the lactose by the two enzyme preparations compared with 35% in control yogurt at 8 h. Lactose was completely hydrolyzed with 93 units enzyme/100 ml in the same period. With 4.7 units/100 ml from our preparation, the oligosaccharide percentage reached its maximum value of 15% of the total sugar in 2 h, slightly decreased to 12% after 8 h, and was 7% after 10 d storage at 5°C. The oligosaccharide percentage with 4.7 units/100 ml from the other preparation was 4 to 8% of the total sugar during fermentation and storage. The oligosaccharide content in low lactose yogurt was 4 to 19 times higher than that obtained with the control yogurt (.3 to 1.7%).