Co-production of Lactic Acid and <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> Cells from Whey Permeate with Nutrient Supplements

Lactic acid and cell production from whey permeate by Lactobacillus rhamnosus with different nutrient supplements were investigated in this study. Yeast extract was identified as the most effective nutrient affecting lactic acid production. Increase in inoculum size from 0.05% to 1% (v/v) resulted in a substantial increase in lactic acid productivity from 0.66 to 0.83 g L⁻¹ h⁻¹ (P < 0.001). The optimal temperature for lactic acid production was 37 °C, while the highest cell production was obtained at 42 °C. When whey permeate and yeast extract concentrations were 6.8% (w/v) and 3 g L⁻¹, respectively, lactic acid productivity reached 0.85 g L⁻¹ h⁻¹ after 48-h cultivation, which is 3.40 times of those without nutrient supplements.     

Effect of dietary levels of fat, α-tocopherol and astaxanthin on colour and lipid oxidation during storage of frozen rainbow trout (Oncorhynchus mykiss) and during chill storage of smoked trout

 Female rainbow trout (Oncorhynchus mykiss) with an initial weight of 0.8–0.9 kg were raised in two experiments including a total of 2550 fish divided into 17 groups. The fish were raised for 6 months on 13 different feeds (four fish groups were replicates) varying in dietary levels of fat (27% or 32%), astaxanthin (40, 70 or 100 mg astaxanthin/kg feed) and vitamin E (α-tocopherol; 100, 300 or 600 mg all-rac-α-tocopheryl acetate/kg feed). The levels of fat, astaxanthin and α-tocopherol in the fillets all increased with increasing dietary levels of each feed component. Furthermore, astaxanthin deposition was found to be significantly improved by increasing the dietary fat level from 27% to 32%, but was not affected by dietary levels of α-tocopherol. The highest deposition of α-tocopherol was found in fish fed the lowest level of astaxanthin (40 mg/kg), whereas α-tocopherol deposition was unaffected by the dietary fat level. Frozen storage (–28  °C) of gutted, cleaned and glazed raw fish for 18 months significantly reduced astaxanthin and α-tocopherol levels, while lipid oxidation, measured as thiobarbituric acid reactive substances (TBARS) was limited. In the first experiment, the highest TBARS levels were found during frozen storage in fish fed the lowest level of astaxanthin (40 mg/kg versus 70 mg/kg or 100 mg/kg); unaffected by dietary levels of α-tocopherol (100 mg/kg versus 600 mg/kg), whereas the dietary astaxanthin level (70 mg/kg versus 100 mg/kg) did not influence lipid oxidation in frozen fish in the second experiment. After brine injection, fillets of fish were smoked and a vacuum-packed (95%), sliced product in a transparent laminate was produced. The quality (pigmentation and lipid oxidation) during 3 weeks of illuminated, chill storage (3  °C) was compared for smoked products produced from fresh fish and from fish stored at –28  °C for 12 months and 18 months. Smoked fillets from fish fed 32% fat were found to be less red than those from fish fed 27% fat, and the astaxanthin content and surface redness of the smoked product decreased during chill storage. Lipid oxidation was pronounced in smoked trout, but a high level of α-tocopherol in the fillet significantly reduced lipid oxidation during chill storage of the smoked product. Lipid oxidation in smoked fillets from fish fed 32% fat was more pronounced than in fish fed 27% fat, but increasing the dietary α-tocopherol level from 300 mg/kg feed to 600 mg/kg feed effectively counteracted the negative effect of the high-fat diet on lipid oxidation in the smoked product. Astaxanthin did not affect lipid oxidation in the chill-stored smoked product, in contrast to the frozen, raw fish. Astaxanthin seems to protect against the very early stages of lipid oxidation, while α-tocopherol is more important as an antioxidant at more advanced stages of lipid oxidation. Received: 8 January 1998 / Revised version: 23 March 1998     

Gilthead seabream (Sparus aurata): suitability for freezing and commercial alternatives

The quality of portion‐size farmed gilthead seabream (Sparus aurata) during frozen storage and the influence of post‐mortem treatments were studied in order to find new ways of marketing this species. Portion‐sized gilthead seabream, fasted for 48 h prior to slaughter, were frozen and stored at ?20 °C for up to one year. Whole fish were frozen immediately after rigor mortis; gutted fish were frozen immediately after rigor and after 5 days of storage in ice. All lots were stored at ?20 °C for up to one year. The myofibrillar protein of this species was very stable and a slight decrease of solubility in salt solutions was found only after one year of frozen storage. A slight decrease in water‐holding capacity and a slight increase in shear strength were observed, but these were lower than in other species. These changes were reflected as increased toughness and reduced juiciness in sensory analysis of the cooked fillets after one year. The main changes in the cooked fillets were observed in odour and flavour. No significant detrimental effect due to the guts was detected during frozen storage. Storage in ice prior to freezing was reflected in sensory assessment of the raw fish, mainly in terms of initial higher demerit points for fishy odour, gills and eyes; however, no effect was observed in the cooked fillets. Copyright © 2004 Society of Chemical Industry     

Assessment of shelf-life of maricultured gilthead sea bream (Sparus aurata) stored in ice

Gilthead sea bream ( Sparus aurata ) were stored in melting ice (0 °C) for a period of 24 days from the time of harvest with sensory assessments of the whole raw fish and of the cooked fish flesh conducted at regular intervals. The ungutted fish was given an EC freshness grade E for up to 3 days, grade A for a further 7 days, and grade B for 4 more days after which it was graded as C (unfit). The sensory score for flavour of the cooked fillets decreased linearly with period of storage: fresh characteristic flavours were present for 2–4 days, decreasing to a relatively bland flavour after 10–12 days. Off flavours were evident by 13–15 days storage and by 18–19 days the flesh was unpalatable. With the possible exception of hypoxanthine, none of the chemicals investigated was particularly useful as an indicator of change. Changes in pH, trimethylamine and total volatile bases during the first half of the edible storage life were insignificant. Deterioration of flesh lipids, assessed by free fatty acid content and thiobarbituric acid value, appeared to present no serious problem during shelf-life. Proximate composition and sensory attributes, appropriate for routine inspection of gilthead sea bream were also determined.     

Oxidation in fish-oil-enriched mayonnaise

 Oxidative protection of mayonnaises with 16% fish oil was studied during cold storage (5  °C) after supplementation with different tocopherol systems: the ternary antioxidant system ascorbic acid, lecithin and tocopherol (A/L/T), and two commercial mixtures, an oil-soluble (Toco 70) preparation and a water-soluble (Grindox 1032) preparation. The physical structure of the fish-oil-enriched mayonnaise was manipulated by adding extra emulsifier (Panodan TR) with the purpose of investigating whether or not this affected the antioxidative activity of the tocopherol mixtures. A number of different analytical techniques HPLC (high-performance liquid chromatography, gas chromatography mass spectrometry (GC-MS), sensory analysis, confocal laser scanning microscopy and rheological measurements) were employed to elucidate the chemical, sensory, structural and rheological aspects of the oxidation process. Discriminant partial least squares regression was used to analyse the data obtained. The three tocopherol preparations not only affected the oxidative stability of the mayonnaises differently, they also influenced the rheological and structural properties of the mayonnaises in different ways. The rheological and structural properties of the mayonnaise were also affected by the addition of extra emulsifier, but this did not influence the formation of fishy and rancid off-flavours. Addition of the A/L/T system caused the immediate formation of distinct fishy and rancid off-flavours in the fresh mayonnaises. The volatile compounds trans-2-heptenal, 4-octen-3-one, 1-octen-3-ol, trans,cis-2,4-heptadienal, trans,trans-2,4-heptadienal, trans-2-octenal, nonanal and trans,cis-2,6-nonadienal were thought to contribute to the fishy and rancid flavours. Addition of Toco 70 did not affect the sensory perception of mayonnaise nor the development of volatile off-flavour compounds as evaluated by GC-MS, but the peroxide values were slightly increased in mayonnaise containing Toco 70 as compared to the other mayonnaises. Mayonnaise with Grindox 1032 seemed to have fewer fishy and rancid off-flavours than mayonnaises without antioxidant. This flavour-protective effect of Grindox 1032 was correlated to an increase in the size of the droplet diameter of mayonnaises supplemented with Grindox 1032. Received: 12 April 1999 / Revised version: 11 June 1999     

Shelf Life of Pretreated Dried Tomato

Determining the shelf life has become a factor of major importance in the development of foods designed to meet consumer demands in terms of quality and safety. The goal of the present study was to investigate the shelf life of vacuum-packed dried tomatoes, stored at both room and refrigeration temperature (4 °C) for a period of 180 days. The following determinations were performed during the storage period studied: microbiological analysis, instrumental color, lycopene, and ascorbic acid. Sorption isotherms were determined at both temperatures (room temperature and 4 °C). The microbiological quality of vacuum-packed dried tomatoes remained unchanged during 180 days for the refrigerated samples and 90 days for the samples stored at room temperature. The rate constant (k) of lycopene degradation of the refrigerated samples and the samples stored at room temperature was 3.209 × 10⁻⁵ and 12.994 × 10⁻⁵/day, respectively. The rate constant (k) of ascorbic acid degradation was 3.339 × 10⁻⁴/day for cold storage and 76.655 × 10⁻⁴/day for storage at room temperature. The tomatoes stored at room temperature were subjected to analysis over 90 days of storage, period after which both the appearance and sensory characteristics of the product fell below the levels of consumer acceptability. As for the tomatoes stored at refrigeration temperature, the original sensory characteristics were maintained throughout the entire storage period of 180 days.     

Quality of Atlantic Mackerel (Scomber scombrus L.) Fillets during Modified Atmosphere Storage

Sensory, physical, biochemical and microbial quality were investigated in mackerel fillets stored under CO₂ modified atmosphere packaging (MAP) at -2°C for 21 days. Packaging gas composition remained constant during storage. An experienced sensory panel found seaweedy and cucumber-like odors (raw) decreased whereas sourness corresponded with lactic acid bacteria increase. Both sensory (raw) fillet ‘yellowness’ and Hunter ‘b’ values increased. Visual (cooked) moistness and expressible fluid decreased but no difference was found in cooking losses. Small increases in total volatile base-nitrogen and trimethylamine-nitrogen paralleled aerobic plate counts; ethanol increased linearly. Coliforms, yeasts and molds were detected at < 10 CFU/g. CO₂ MAP storage at – 2°C can maintain Grade 1 shelf-life extension for ≥ 21 days     

Changes in biogenic amines of silver carp (Hypophthalmichthys molitrix) fillets stored at different temperatures and their relation to total volatile base nitrogen,microbiological and sensory score

BACKGROUND: Biogenic amines have received considerable attention owing to their undesirable effects in humans. There are few studies of changes in biogenic amine contents related to freshwater fish. Silver carp is an important freshwater fish species in China. This study aimed to investigate the changes in biogenic amines and their relation to total volatile base nitrogen (TVB‐N), microbiological and sensory score of silver carp fillets stored at 0, 3 and 15 °C. RESULTS: The total biogenic amine contents of all silver carp fillets (regardless of storage time and temperature) ranged from 13.05 to 318.10 mg kg^?1. Putrescine and histamine were the main biogenic amines in silver carp fillets during storage. Cadaverine was only detected after 12 days at 3 °C and after 2 days at 15 °C. Spermidine and spermine contents increased during the early storage period and then slightly decreased. CONCLUSION: Low temperature could control the quality of silver carp fillets by inhibiting the contents of biogenic amines. Putrescine showed significant correlation with TVB‐N, total aerobic content, sensory score, tryptamine and phenylethylamine. Putrescine was a good quality marker of silver carp fillets in the cold chain. Copyright © 2012 Society of Chemical Industry     

Effect of previous frozen storage on quality changes of grass carp (Ctenopharyngodon idellus) fillets during short‐term chilled storage

The effect of chilled, frozen and freeze‐chilled storage on quality of grass carp fillets and soups was evaluated by sensory score, total aerobic counts and biochemical quality. Fish fillets were stored at 4 °C for 6 days (T1), ?40 °C for 12 h and then at ?20 °C for 5 days (T2), ?40 °C for 12 h and then at ?20 °C for 5 days, followed by at 4 °C for 4 days (T3). T1 showed higher sensory score, salt‐soluble protein content, better colour and texture qualities than T2 and T3 within 3 days. All fillets kept good quality based on the acceptable limit of sensory score, total volatile basic nitrogen and total aerobic count during storage time. According to the transportation and retail time, chilled storage is appropriate when it is within 3 days. If it extends for 5 days, freeze‐chilling treatment keeps better quality, but later chilled fillets should be retailed within 4 days.     

Chemical,microbiological and sensory changes in thawed frozen fillets of sardine (Sardina pilchardus) during marination

Frozen fillets of sardine (Sardina pilchardus) were used to make marinades. The marinating process was performed in 7% acetic acid and 14% sodium chloride in barrels. The fish : solution ratio was (1.5:1). The marination was carried out at 4 °C until the end point of marinating was assessed by sensory texture analysis. The complete marinating of sardine fillets required 22 days at 4 °C. According to statistical analysis at the beginning and at the end of the storage in barrels there were no significant differences (p>0.05) in chemical analytical results. After the sardine fillets were put into the barrels, total viable count, lactic acid bacteria count, psychrotrophic bacteria count, yeast and mould counts were reduced.     

The Effect of Thyme and Garlic Oil on the Preservation of Vacuum-Packaged Hot Smoked Rainbow Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

Quality and shelf life of filleted hot smoked rainbow trout packaged in vacuum packaging (untreated) and with thyme oil (TO), and garlic oil (GO)-treated, after vacuum-packaged (VP) conditions stored at 2 °C, were investigated by measurement of microbiological, sensory and physicochemical analyses. Acceptability scores for appearance, odour, taste and texture of untreated and treated smoked trout decreased with storage time. The limit of sensory acceptance was reached after 5 weeks for the untreated samples, after 7 weeks for with thyme oil-treated samples (TO + VP) and after 6 weeks for with garlic oil-treated (GO + VP) samples. Total viable count evaluated that thyme oil- and garlic oil-treated vacuum-packaged samples had good results up to 7 weeks compared with 5 weeks in control vacuum packs. Psychrotrophic counts for untreated and treated samples exceeded 6 log cfu/g after 5 and 6 weeks of cold storage, respectively. Treated samples presented the lowest lactic acid bacteria counts. Total volatile basic nitrogen and trimethylamine nitrogen values gave acceptable results for up to 5 weeks for the untreated samples, 7 weeks for thyme- and garlic oil-treated samples. Treated with thyme oil samples presented the lowest thiobarbituric acid index values. The results obtained from this study showed that the shelf life of hot smoked rainbow trout stored in cold storage (2 °C), as determined by overall acceptability of all data, is 5 weeks for vacuum-packaged samples and 7 weeks for thyme- and garlic oil-treated vacuum-packaged smoked fish.     

Comparison of the quality of sous-vide and conventionally processed carrots

 The quality of "sous-vide" and conventionally processed carrots was evaluated by sensory analysis. Furthermore, changes in colour, odour, taste and texture of the "sous-vide" products during storage over 21 days at 2  °C were determined with instrumentation. According to the panel's evaluation, the sensory quality of the "sous-vide" processed carrots remained almost constant over the storage period. However, the shear force values indicated a strengthening of the texture. The colour changed from typical orange to red-orange. A slight decrease of the flavour and aroma intensity was recorded by the panel, which was confirmed by the data gained with the Electronic nose and the AromaScan. The sous-vide processed products were always preferred and therefore obtained a higher valuation even at the end of storage. In contrast to the conventionally processed carrots, only a slight reduction in the content of sucrose, fructose and glucose was determined for the sous-vide product. Received: 3 February 1998 / Revised version: 17 April 1998     

Comparison of the quality of sous-vide and conventionally processed carrots

 The quality of "sous-vide" and conventionally processed carrots was evaluated by sensory analysis. Furthermore, changes in colour, odour, taste and texture of the "sous-vide" products during storage over 21 days at 2  °C were determined with instrumentation. According to the panel's evaluation, the sensory quality of the "sous-vide" processed carrots remained almost constant over the storage period. However, the shear force values indicated a strengthening of the texture. The colour changed from typical orange to red-orange. A slight decrease of the flavour and aroma intensity was recorded by the panel, which was confirmed by the data gained with the Electronic nose and the AromaScan. The sous-vide processed products were always preferred and therefore obtained a higher valuation even at the end of storage. In contrast to the conventionally processed carrots, only a slight reduction in the content of sucrose, fructose and glucose was determined for the sous-vide product. Received: 3 February 1998 / Revised version: 17 April 1998     

Effect of frozen storage on the quality of whole fish and fillets of horse mackerel (Trachurus trachurus) and mediterranean hake (Merluccius mediterraneus)

 Whole fish and fillets of horse mackerel (Trachurus trachurus) and mediterranean hake (Merluccius mediterraneus) were assessed for quality (physical, chemical and sensory attributes) changes throughout 12 months of frozen storage at −18 °C. The pH, expressible water (EXW), quantities of trimethylamine (TMA), dimethylamine (DMA), formaldehyde (FA), the total volatile base nitrogen (TVB-N) the thiobarbituric acid number (TBA), peroxide value (PV) and amount of free fatty acids (FFA) increased, while sensory attributes (odour, taste, texture) decreased during the frozen storage period. A comparison of quality scores between whole fish and fillets of horse mackerel and mediterranean hake showed that there were no significant differences (P>0.05) in attribute scores. There were, however, significant differences (P<0.05) in pH, EXW, TMA, DMA, FA, TVB-N, TBA, FFA and PV. Received: 19 April 1996/Revised version: 7 September 1996     

Effect of pH on fluorescence formation related to fish deterioration

 Earlier studies have investigated fluorescence at different excitation/emission maxima during common types of fish processing. A shift towards higher wavelength maxima was observed and measured as the ratio between two of the maxima tested. This fluorescence ratio (δF) correlated with increased fish damage. The present work is focused on the effect that pH can have on the formation of fluorescent compounds in fish muscle systems. Minced hake muscle was homogenised with 0.1 M phosphate buffer of different pH values (5, 6, 7 and 8) and was stored at 30  °C for up to 30 days. Lipid damage, measured as the δF value of the aqueous reaction medium and the fish lipid extract, indicated little difference between the effect of different pH values under the conditions employed in the present experiment. The effetct of formaldehyde (FA) in the same reaction medium was also evaluated. It was observed that FA had a positive effect on the fluorescence shift occurring in the aqueous reaction medium, so that a higher δF value was observed for pH 7 and pH 8, especially for the latter. It is concluded that changes in the δF value during fish storage and/or processing are of special interest as, in addition to FA formation and lipid oxidation, significant pH increases are expected to occur as a result of damage. Received: 2 February 1998 / Revised version: 6 April 1998     

The impact of strawflower and mistletoe extract on quality properties of rainbow trout fillets

Many plants, including strawflower and mistletoe, contain antioxidants and antimicrobials, which can increase the shelf life of seafood. The main aim of this study was to investigate the effect of mistletoe and strawflower extracts at doses of 0.5% (w/v) on the sensory, chemical and microbiological properties of rainbow trout fillets during 27 days of storage at 2 ± 1 °C. The phenolic compounds in these plants have been studied, but their effects on food quality and storage properties have not been reported. We found that extract of mistletoe did not extend the shelf life of the fillets; however, the strawflower extract show high antimicrobial activity in fish fillets. The shelf life of rainbow trout was 20 days for the control and fish treated with mistletoe extract and 23 days for fish treated with strawflower extract. The antioxidant effect of extracts on fish fillets was weak, whereas strawflower extract had high antimicrobial effect. Peroxide value (PV) and thiobarbituric acid (TBA) values fluctuated during storage periods around main values below 14 meq O₂ kg^?1 and 0.6 MA kg^?1, respectively. Putrescine, cadaverine, spermine, spermidine, serotonin, tyramine and dopamine were main amine, whereas histamine accumulated at low levels (<2 mg per 100 g). Strawflower suppressed biogenic amine accumulation in fish fillets. At the limit of acceptability, total viable count (TVC), Enterobacteriaceae and lactic acid bacteria count remained below 7.6, 6.83 and 8.01 log CFU g^?1, respectively. The results of this study show that ethanol extracts of strawflower improve the shelf life of rainbow trout.     

贮藏条件对红薯叶酸菜品质的影响

目的:探寻乳酸菌发酵红薯叶酸菜的贮藏条件及贮藏期间品质变化规律。方法:真空包装后以常温25℃、低温4℃、80℃—15 min巴氏杀菌后4℃、85℃-10 min巴氏杀菌后4℃贮藏,比较4个条件下红薯叶酸菜pH值、乳酸含量、亚硝酸盐含量、菌落数、色差值及感官评分。结果:25℃下贮藏亮度下降最快,第60天色泽明显变深;4℃下贮藏第30天的酵母菌数量最多,第45天的乳酸菌数量最多;80℃-15 min杀菌后贮藏第90天的大肠杆菌数量最少,感官评分达到食用临界值;85℃-10 min杀菌后贮藏第75天的总菌数量最少,感官评分及格。pH值与亚硝酸盐含量呈极显著负相关,与L值、感官评分呈极显著正相关;乳酸菌数与酵母菌、大肠杆菌、总菌数呈极显著正相关,其中感官评分在第一主成分中的载荷量最大。结论:80℃-15 min杀菌后4℃贮藏,其主要微生物为乳酸菌,其他指标均符合安全标准,感官品质稳定,更适合作为酸菜的贮藏条件,贮藏期可达90 d。     

Improving the keeping quality of pomegranate fruit by intermittent warming

 Mollar pomegranates (Punica granatum, Punicaceae) were stored at 0  °C or 5  °C and 95% relative humidity (RH) for 80 days. Intermittent warming (IW) cycles of 1 day at 20  °C every 6 days, during which time the fruit had been stored at 0  °C or 5  °C, followed by a shelf-life period of 7 days at 15  °C and 70% RH were applied. IW during storage at 0  °C was the best treatment for maintaining the red skin colour as at harvest. However the red colour of the arils (hue angle) was kept better under warming at 5  °C. Although significant changes in the individual anthocyanins were observed in all treatments, particularly after the shelf-life period, the total anthocyanin content of the juice at harvest was maintained. While storage at 0  °C avoided decay although increased the risk of chilling injuries such us pitting and husk scald, 5  °C-storage reduced these injuries but fungal attacks were not inhibited. After the shelf-life period, IW alleviated chilling injuries without any incidence of decay. Warming treatments gave very good results with respect to storage and the keeping quality of pomegranates, particularly when applied during storage at 0  °C. Received: 10 February 1998 / Revised version: 24 April 1998     

Modeling Microbial Spoilage and Quality of Gilthead Seabream Fillets: Combined Effect of Osmotic Pretreatment,Modified Atmosphere Packaging,and Nisin on Shelf Life

ABSTRACT: The objective of the study was the kinetic modeling of the effect of storage temperature on the quality and shelf life of chilled fish, modified atmosphere-packed (MAP), and osmotically pretreated with the addition of nisin as antimicrobial agent. Fresh gilthead seabream (Sparus aurata) fillets were osmotically treated with 50% high dextrose equivalent maltodextrin (DE 47) plus 5% NaCl. Water loss, solid gain, salt content, and water activity were monitored throughout treatment and treatment conditions were selected for the shelf life study. Untreated and osmotically pretreated slices with and without nisin (2 × 10⁴ IU/100 g osmotic solution), packed in air or modified atmosphere (50% CO₂–50% air), and stored at controlled isothermal conditions (0, 5, 10, and 15 °C) were studied. Quality assessment and modeling were based on growth of several microbial indices, total volatile nitrogen, trimethylamine nitrogen, lipid oxidation (TBARS), and sensory scoring. Temperature dependence of quality loss rates was modeled by the Arrhenius equation, validated under dynamic conditions. Pretreated samples showed improved quality stability during subsequent refrigerated storage, in terms of microbial growth, chemical changes, and organoleptic degradation. Osmotic pretreatment with the addition of nisin in combination with MAP was the most effective treatment resulting in significant shelf life extension of gilthead seabream fillets (48 days compared to 10 days for the control at 0 °C).     

Lactic Acid Concentration and Microbial Spoilage in Anaerobically and Aerobically Stored Ground Beef

Lactic acid concentration of coarsely ground beef, packaged in low oxygen permeability casings, is inversely related to sensory acceptability of the meat following regrinding and aerobic storage for six additional days. Samples, reground and stored aerobically after periods of anaerobic storage, were assayed for total aerobic bacterial counts, percentage of gram-positive microflora, and pH. Lactic acid concentration ranged from 130 to 1055 mg per 100g ground beef and increased with anaerobic storage. During 6 days of aerobic storage, both increases and decreases in concentration were noted and related to changes in microbial flora. A positive correlation was found between the proportion of gram-positive bacteria (lactic acid bacteria) and lactic acid and pH.