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PubMed1987

Pediococci and biotechnology.

Nomenclature changes of pediococci postdate the publication of Bergey's Manual. Pediococci possess both a "group" and a "type" antigen. They are gram positive, asporogenous, nonmotile, generally catalase negative, but may possess catalase-like activity. The pediococci may have either a cytochrome or a flavoprotein enzyme system. Anaerobically they are homofermentative using the PEP:PTS and the EMP pathway. Catalase positive strains utilize glucose aerobically and anaerobically while lactose and glycerol are only used aerobically. Some pentoses are fermented to lactate and acetate. Absolute requirement for folinic acid and nearly all amino acids is observed. Pediococci grow luxuriously in All Purpose Tween (APT) broth and are isolated on Rogosa SL agar. Detection can be done by electrical impedance and fluorescent antibody techniques. The Arrhenius concept was utilized in selecting metabolically efficient strains. Antibiotics, antioxidants, some chloride salts and some spices are detrimental to the pediococci. On the other hand, some chloride salts, manganese, and some spices are stimulant. Dialysis-fermentation and immobilization of pediococcal cells were recorded. Some strains decarboxylate histidine to histamine. The resting cell metabolism and the production of bacteriocin have been utilized in antibiosis. An intra and intergeneric genetic transfer system of plasmids from pediococci was by a conjugation-like mechanism. Formation of bacteriocin and fermentation of carbohydrates were linked to plasmids. Lytic bacteriophages to pediococci have not yet been identified. Industrial cultures are mainly frozen concentrates. Linear equations were developed to model the fermentative activity of pediococci and the effects of environmental factors.

Antibiosis↗
PubMed2004

The passage of lactic acid bacteria from silage into rumen fluid, in vitro studies.

Inoculated silages sometimes improve cattle performance, possibly because of probiotic effects of lactic acid bacteria (LAB) silage inoculants. The cause of improved animal performance following feeding with inoculated silage is unclear. One issue in studying this phenomenon is to find out whether LAB pass from silage into the rumen fluid and survive in it. The purpose of the present study was to determine whether LAB from inoculated and uninoculated silages pass into the rumen fluid in vitro. Wheat and corn silages, uninoculated or inoculated with 1 of 10 commercial silage inoculant LAB, were prepared in glass jars. After ensiling, a 2.5-g silage sample was added to 25 mL of heat-sterilized or strained rumen fluid together with 5 g/L glucose, and incubated for 48 h at 39 degrees C. Analysis of the incubated rumen fluid included pH measurement, enumeration of LAB, and determination of lactic acid and volatile fatty acids (VFA). The pH of the rumen fluid decreased during incubation; both heat-sterilized and strained rumen fluid contained large numbers of LAB. The heat-sterilized rumen fluid contained lactic acid in addition to VFA, whereas the strained rumen fluid contained only VFA. The results indicate that LAB pass from silage samples into the rumen fluid in vitro and survive there. Their interactions with rumen microorganisms should be studied further to understand how some silage inoculant LAB exhibit probiotic effects in dairy cattle.

Animals↗
PubMed2004

The influence of treatment with dual purpose bacterial inoculants or soluble carbohydrates on the fermentation and aerobic stability of bermudagrass.

This study determined the effectiveness of an inoculant (BB), molasses, or a mixture of either BB and molasses (BBM) or BB and fibrolytic enzymes (BBE) for improving the fermentation and aerobic stability of bermudagrass. A 6-wk regrowth of Tifton 85 bermudagrass was conserved in quadruplicate mini silos alone or after treatment application. The inoculant contained a mixture of P. pentosaceus 12455, 1 x10(5) cfu/g of fresh forage, L. buchneri 40788, 4 x10(5) cfu/g of fresh forage, and beta-glucanase, alpha-amylase, and xylanase; BBE contained similar bacteria and enzymes as BB, but greater enzyme activities. Chemical composition was quantified after 2, 4, 7, 30, and 60 d of ensiling. Microbial composition and aerobic stability were measured after 60 d of ensiling, at which point the pH of additive-treated silages was consistently lower and DM recovery was higher than in untreated silages. The BB, BBM, and molasses-treated silages had less ammonia N than untreated silages, and BB, BBM, and BBE-treated silages had less residual water-soluble carbohydrates than untreated silages. All silages had high acetic acid (47.5 g/kg DM) and low lactic acid (1.7 g/kg DM) concentrations. However, untreated and BBE-treated silages had more butyric acid and ammonia N, suggesting that a clostridial fermentation had occurred. These butyric forages were more aerobically stable (27 d) but less desirable for feeding than those ensiled with BB or molasses, which were stable for 6.9 d. In conclusion, BB and molasses treatments improved the digestibility and fermentation of bermudagrass and produced higher quality silages that were stable for 6.9 d. Mixing BB with molasses or the inoculant tested was not more beneficial than BB or molasses alone.

Acetic Acid↗
PubMed2006

Nonstarter lactic acid bacteria biofilms and calcium lactate crystals in Cheddar cheese.

A sanitized cheese plant was swabbed for the presence of nonstarter lactic acid bacteria (NSLAB) biofilms. Swabs were analyzed to determine the sources and microorganisms responsible for contamination. In pilot plant experiments, cheese vats filled with standard cheese milk (lactose:protein = 1.47) and ultrafiltered cheese milk (lactose:protein = 1.23) were inoculated with Lactococcus lactis ssp. cremoris starter culture (8 log cfu/mL) with or without Lactobacillus curvatus or Pediococci acidilactici as adjunct cultures (2 log cfu/mL). Cheddar cheeses were aged at 7.2 or 10 degrees C for 168 d. The raw milk silo, ultrafiltration unit, cheddaring belt, and cheese tower had NSLAB biofilms ranging from 2 to 4 log cfu/100 cm2. The population of Lb. curvatus reached 8 log cfu/g, whereas P. acidilactici reached 7 log cfu/g of experimental Cheddar cheese in 14 d. Higher NSLAB counts were observed in the first 14 d of aging in cheese stored at 10 degrees C compared with that stored at 7.2 degrees C. However, microbial counts decreased more quickly in Cheddar cheeses aged at 10 degrees C compared with 7.2 degrees C after 28 d. In cheeses without specific adjunct cultures (Lb. curvatus or P. acidilactici), calcium lactate crystals were not observed within 168 d. However, crystals were observed after only 56 d in cheeses containing Lb. curvatus, which also had increased concentration of D(-)-lactic acid compared with control cheeses. Our research shows that low levels of contamination with certain NSLAB can result in calcium lactate crystals, regardless of lactose:protein ratio.

Animals↗
PubMed1989

Microbial inoculation of alfalfa haylage: ensiling characteristics and milk production response when fed to early lactation dairy cows.

Third-cutting alfalfa hay harvested at bud stage and wilted to approximately 65% moisture was treated with a live bacterial inoculant at the rate of 300,000 cfu/g fresh alfalfa. Treated alfalfa was packed in polyethylene bags. Samples were taken at time of ensiling and d 1, 2, 3, 4, 7, and 28 postensiling. Mean pH was lower in the treated haylage (5.09 and 5.71 for treatment and control, respectively). Mean temperatures were higher in the treated haylage (30.0 and 28.0 degrees C for treatment and control, respectively). Mold count, water-soluble carbohydrate, alpha amino nitrogen, CP, and ADF were not affected by treatment. Regardless of treatment, pH, mold counts, and water-soluble carbohydrates declined with time. There was no significant difference between treatments for DM intake, milk production, and milk composition.

Analysis of Variance↗
PubMed1995

Additives containing bacteria and enzymes for alfalfa silage.

First-cutting alfalfa was wilted, harvested from alternate rows, left untreated or treated with additives containing lactic acid bacteria and enzymes (cellulase, amylase, and pectinase), and ensiled in bag silos. Inoculation increased lactic acid bacteria from 5 x 10(4) to 1 x 10(6) cfu/g of forage. Because treatments were bagged consecutively, the DM of treated silages was higher than that of untreated silage. However, after 4 d of ensiling, the pH of treated silage, about 4.3, was lower than that of untreated silage, 4.7, and remained lower throughout the ensiling period. After 177 d of ensiling, total lactate was about 25% higher, and ammonia N was about 40% lower, in treated silage. In addition, NDF and ADF contents were lower in treated than in untreated silage. Between 51 and 177 d of storage, glucose content increased in treated silage, but not in untreated silage, suggesting that some plant cell-wall hydrolysis occurred during prolonged storage. In vitro digestion of NDF did not differ among treatments during early incubation, but the extent of digestion after 36 and 48 h was lower in treated than in untreated silage. The microbial and enzyme silage additives used in this study improved fermentation characteristics and reduced fiber content of silage but decreased the in vitro digestibility of fiber.

Amylases↗
PubMed1996

Application of direct-fed microbial bacteria and fructooligosaccharides for salmonella control in broilers during feed withdrawal.

Providing direct-fed-microbial (DFM) bacteria and fructooligosaccharides (FOS) for the control of potential escalation of Salmonella colonization during simulated feed withdrawal and confinement was assessed. Eight hundred and eighty broilers (16 pens; 55 chicks per pen) were reared to 6 wk of age. Chicks were sprayed with a solution containing 10(6) nalidixic-acid resistant Salmonella typhimuriumNR cells per milliliter on the 2nd d after hatching. Because this first challenge did not yield a high infection rate, chickens were rechallenged per Os at Day 18 by providing water containing 10(7) cells of S. typhimuriumNR per milliliter. At 3 and 5 wk of age, 10 birds per pen were euthanatized and cecal Salmonella were quantified (log colony-forming units per gram). Feed was removed from all pens at 6 wk, and pens were randomly assigned to be either the treatment group or the control group. The treatment groups were provided a DFM (mixture of nine bacteria) and FOS 50 (R) (10%) in the drinking water. The control groups received drinking water only. After 6 h of feed withdrawal, chickens were cooped (eight per coop) and held 10 h. Immediately after confinement, 10 chickens were used for cecal enumeration of S. typhimuriumNR. Salmonella colonization declined from 99% at 3 wk to 44% at 5 wk. After feed withdrawal, application of the treatment, and confinement, 11 and 14% of the treated and control groups, respectively, yielded S. typhimuriumNR by direct plating from ceca (3.87 and 3.75 log 10 cfu/g, respectively). No difference (P > 0.05) in Salmonella colonization occurred between the treated and the control groups; however, enrichment of ceca (incubation in nutrient broth at 37 C for 24 h) yielded a higher incidence of S. typhimuriumNR in the control groups (32% in the treated vs 51% in the control). Ceca weights were greater in the treated group (P < 0.05). Simulated feed withdrawal and confinement did not escalate Salmonella colonization in the chicken ceca.

Animals↗
PubMed1998

Inactivation of microorganisms with microwaves at reduced temperatures.

We developed a pilot-plant nonthermal flow process using microwave energy to inactivate microorganisms. The process consists of multiple passes through the microwave generator. Each passed material goes to a receiving tank for subsequent passes. The flow rate was 0.96 to 1.26 kg/min and the dwell time per pass was 1.1 to 1.5 min. Five passes were used. The microwave energy is instantaneously and simultaneously applied to the system, and thermal energy is removed by a cooling tube within the process line in the microwave generator. The cooling tube maintains the temperature below 40 degrees C. There was significant reduction in microorganisms in water, 10% glucose solution, and apple juice, and in yeast in beer. There was a slight decrease in microorganisms in tomato juice, pineapple juice, apple cider, and beer; and no effect in skim milk.

Beer↗
PubMedDate not supplied

Enumeration of probiotic pediococci in animal feed: interlaboratory study.

An enumeration method to be used as official control under Council Directive 70/524/EEC for probiotic pediococci used as feed additives was validated for consideration for adoption as Comitée Européen de Normalisation (CEN) and ISO standards. Seventeen laboratories in 11 European countries carried out an interlaboratory study. A spread plate method following BS ISO 15214:1998 using 4 different agars [MRS, acidified MRS, MRS with triphenyl tetrazolium chloride (TTC), and a newly developed pediococci selective medium (PSM)] was validated. Precision data in terms of repeatability (r) and reproducibility (R) of the method for each medium using different feeding stuffs with a high and a low inoculation level were determined. Pediococci were present in the samples in mixtures with other probiotics. The enumeration of pediococci on all agars showed an RSDr value of 0.4-3.1% and an RSDR of 1.3-4.8%. MRS agar was preferred, followed by acidified MRS and MRS + TTC agar. All 4 media gave similar counts. Depending on the presence and concentration of other probiotic, such as enterococci, lactobacilli, and yeast, acidified MRS or MRS + TTC agar are recommended. The PSM was selective for pediococci and can be used if this species is present at a concentration more than 10-fold lower than other species that can grow on the MRS agars. The methodology with all 4 media is not applicable to mineral feed.

Agar↗