Are fructophilic lactic acid bacteria (FLAB) beneficial to humans?

Fructophilic lactic acid bacteria (FLAB) are heterofermentative and related to the genera Fructilactobacillus, Convivina, Leuconostoc, Oenococcus and Weissella. Although they generally prefer fructose above glucose, obligate heterofermentative species will ferment glucose in the presence of external electron acceptors such as pyruvate and fructose. Little is known about the presence of FLAB in the human gut, let alone probiotic properties. In this review we discuss the possible role FLAB may have in the human gastro-intestinal tract (GIT) and highlight the advantages and disadvantages these bacteria may have in individuals with a diet high in fructose.

Does the Future of Antibiotics Lie in Secondary Metabolites Produced by Xenorhabdus spp.? A Review.

The over-prescription of antibiotics for treatment of infections is primarily to blame for the increase in bacterial resistance. Added to the problem is the slow rate at which novel antibiotics are discovered and the many processes that need to be followed to classify antimicrobials safe for medical use. Xenorhabdus spp. of the family Enterobacteriaceae, mutualistically associated with entomopathogenic nematodes of the genus Steinernema, produce a variety of antibacterial peptides, including bacteriocins, depsipeptides, xenocoumacins and PAX (peptide antimicrobial-Xenorhabdus) peptides, plus additional secondary metabolites with antibacterial and antifungal activity. The secondary metabolites of some strains are active against protozoa and a few have anti-carcinogenic properties. It is thus not surprising that nematodes invaded by a single strain of a Xenorhabdus species are not infected by other microorganisms. In this review, the antimicrobial compounds produced by Xenorhabdus spp. are listed and the gene clusters involved in synthesis of these secondary metabolites are discussed. We also review growth conditions required for increased production of antimicrobial compounds.

Effect of a Multi-Species Probiotic on the Colonisation of Salmonella in Broilers.

Newly hatched broiler chickens are the most susceptible to Salmonella infections, especially during the first 24 h. At this age, the gut microbiome is not fully developed and offers little protection in the form of competitive exclusion. In this study, one group of newly hatched, Salmonella-free broilers were colonised with a multi-species probiotic (2.0 × 1010 to 8.9 × 1010 CFU per kg feed) for 28 days, consisting of Lactobacillus crispatus, Lactobacillus salivarius, Lactobacillus gallinarum, Lactobacillus johnsonii, Enterococcus faecalis and Bacillus amyloliquefaciens. Broilers in another group received oxytetracycline (200 mg/kg feed), instead of the probiotic, for 28 days. On days 9 and 10, broilers in both groups were gavaged with 9 × 107 CFU Salmonella Enteritidis A9, a pathogenic strain isolated from infected broilers. On day 14, Salmonella was detected in the ceca of 95% of broilers treated with the multi-species probiotic, but 2 weeks later, almost half of the birds (45%) had no Salmonella in their ceca. Similar results were recorded after 28 days of treatment with oxytetracycline. Only 10% of Salmonella-infected birds not treated were Salmonella-free on day 28. Growth performance, immune organ weight (spleen and bursa of Fabricius) and whole blood cell counts of birds treated with the multi-species probiotic and oxytetracycline, respectively, were similar to untreated and uninfected birds throughout the 28-day trial (p > 0.05). On day 14, serum lysozyme levels of broilers exposed to the probiotic were lower (8.0 µg/mL) compared with those of broilers treated with oxytetracycline (11.0 µg/mL). Although the multi-species probiotic and oxytetracycline stimulated the immune system, probiotics are safer to use than antibiotics and should be the preferred choice of treatment.

Differentiation between Bacillus amyloliquefaciens and Bacillus subtilis isolated from a South African sugarcane processing factory using ARDRA and rpoB gene sequencing.

A total of 104 exopolysaccharide (gum)-producing bacteria were isolated from the juice screen and juice tank in a sugarcane processing factory at times of low- and high dextran concentrations in the produced sugar. Dextran is an indicator of cane deterioration and sucrose loss after harvesting of the cane. The isolates were identified as Bacillus amyloliquefaciens (96 isolates) and Bacillus subtilis (eight isolates) based on restriction enzyme banding patterns of amplified 16S rRNA genes and rpoB gene sequence analysis. Exopolysaccharide production in sugarcane is normally associated with dextran produced by Leuconostoc mesenteroides. B. amyloliquefaciens, and to a lesser extent B. subtilis, could, however, also be responsible for exopolysaccharide (slime or gum) production in cane processing factories.

Xenorhabdus khoisanae SB10 produces Lys-rich PAX lipopeptides and a Xenocoumacin in its antimicrobial complex.

BACKGROUND: Xenorhabdus spp. live in close symbiosis with nematodes of the Steinernema genus. Steinernema nematodes infect an insect larva and release their symbionts into the haemocoel of the insect. Once released into the haemocoel, the bacteria produce bioactive compounds to create a semi-exclusive environment by inhibiting the growth of bacteria, yeasts and molds. The antimicrobial compounds thus far identified are xenocoumacins, xenortides, xenorhabdins, indole derivatives, xenoamicins, bicornutin and a number of antimicrobial peptides. The latter may be linear peptides such as the bacteriocins xenocin and xenorhabdicin, rhabdopeptides and cabanillasin, or cyclic, such as PAX lipopeptides, taxlllaids, xenobactin and szentiamide. Thus far, production of antimicrobial compounds have been reported for Xenorhabdus nematophila, Xenorhabdus budapestensis, Xenorhabdus cabanillasii, Xenorhabdus kozodoii, Xenorhabdus szentirmaii, Xenorhabdus doucetiae, Xenorhabdus mauleonii, Xenorhabdus indica and Xenorhabdus bovienii. Here we describe, for the first time, PAX lipopeptides and xenocoumacin 2 produced by Xenorhabdus khoisanae. These compounds were identified using ultraperformance liquid chromatography, linked to high resolution electrospray ionisation mass spectrometry and tandem mass spectrometry. RESULTS: Cell-free supernatants of X. khoisanae SB10 were heat stable and active against Bacillus subtilis subsp. subtilis, Escherichia coli and Candida albicans. Five lysine-rich lipopeptides from the PAX group were identified in HPLC fractions, with PAX1' and PAX7 present in the highest concentrations. Three novel PAX7 peptides with putative enoyl modifications and two linear analogues of PAX1' were also detected. A small antibiotic compound, yellow in colour and λmax of 314 nm, was recovered from the HPLC fractions and identified as xenocoumacin 2. The PAX lipopeptides and xenocoumacin 2 correlated with the genes and gene clusters in the genome of X. khoisanae SB10. CONCLUSION: With UPLC-MS and MSe analyses of compounds in the antimicrobial complex of X. khoisanae SB10, a number of PAX peptides and a xenocoumacin were identified. The combination of pure PAX1' peptide with xenocoumacin 2 resulted in high antimicrobial activity. Many of the fractions did, however, contain labile compounds and some fractions were difficult to resolve. It is thus possible that strain SB10 may produce more antimicrobial compounds than reported here, as suggested by the APE Ec biosynthetic complex. Further research is required to develop these broad-spectrum antimicrobial compounds into drugs that may be used in the fight against microbial infections.

Bacteriocin production and adhesion properties as mechanisms for the anti-listerial activity of Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA.

Probiotics play an important role in maintaining a healthy and stable intestinal microbiota, primarily by preventing infection. Probiotic lactic acid bacteria (LAB) are known to be inhibitory to many bacterial enteric pathogens, including antibiotic-resistant strains. Whilst the positive role that probiotics have on human physiology, specifically in the treatment or prevention of specific infectious diseases of the gastro-intestinal tract (GIT) is known, the precise mechanistic basis of these effects remains a major research goal. In this study, molecular evidence to underpin the protective and anti-listerial effect of Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA against orally administered Listeria monocytogenes EGDe in the GIT of mice is provided. Bacteriocins plantaricin 423 and mundticin ST4SA, produced by L. plantarum 423 and E. mundtii ST4SA, respectively, inhibited the growth of L. monocytogenes in vitro and in vivo. Bacteriocin-negative mutants of L. plantarum 423 and E. mundtii ST4SA failed to exclude L. monocytogenes EGDe from the gastrointestinal tract (GIT) of mice. Furthermore, L. plantarum 423 and E. mundtii ST4SA failed to inhibit recombinant strains of L. monocytogenes EGDe in vivo that expressed the immunity proteins of the two bacteriocins. These results confirmed that bacteriocins plantaricin 423 and mundticin ST4SA acted as anti-infective mediators in vivo. Compared to wild type strains, mutants of L. plantarum 423 and E. mundtii ST4SA, in which the adhesion genes were knocked out, were less effective in the exclusion of L. monocytogenes EGDe from the GIT of mice. This work demonstrates the importance of bacteriocin and adhesion genes as probiotic anti-infective mechanisms.

Polyacrylonitrile (PAN) nanofibres spun with copper nanoparticles: an anti-Escherichia coli membrane for water treatment.

Copper nanoparticles (CNPs) were mixed with polyacrylonitrile (PAN) and electrospun into nanofibres (CuPAN nanofibres). PAN nanofibres containing 1.0, 3.0 and 5.0% copper (w/v) displayed beads-on-string morphology with protrusions of copper particles. The diameter of the CuPAN nanofibres differed according to the copper content, ranging from 386 nm (1.0%, w/v, copper) to 922 nm (5.0%, w/v, copper). No chemical interaction of copper with PAN was observed when studied with X-ray diffraction, ATR-FTIR (attenuated total reflection-Fourier transform infrared) spectroscopy and TGA (thermogravimetric analysis). None of the CuPAN nanofibres showed signs of degradation after 7 days in water. Bacteria suspended in random mobility buffer and filtered through a 3% CuPAN nanofibre membrane (25 mm diameter, 75-80 µm thickness), at a filtration rate of 20 ml min-1, reduced the cell numbers of enterotoxigenic Escherichia coli (ETEC) from 3.3 × to 2.1 × 106 cfu ml-1 and methicillin-resistant Staphylococcus aureus (MRSA) from 1.2 × 10 to 1.3 × 103 cfu ml-1. Membranes produced with 1.0, 3.0 and 5.0% (w/v) CuPAN inhibited the growth of enteroaggregative E. coli (EAEC), enterohemorrhagic E. coli (EHEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), ETEC and MRSA, as shown with LIVE/DEAD™ BacLight™ staining. Real-time bactericidal activity of CuPAN membranes was recorded by staining the cells with SYTO 9 and PI, followed by flow cytometry. Filter membranes made from CuPAN fibres may be used to reduce pathogenic E. coli cell numbers in potable water.

Our gut microbiota: a long walk to homeostasis.

The microbiome of the human gastrointestinal tract (GIT) consists of billions of bacteria, fungi and viruses, of which bacteria play the most important role in nutrition, immune development, production of vitamins and maintaining a well-balanced (homeostatic) microbial population. Many papers have been published on the microbiota in the human GIT, but little is known about the first group of bacteria that colonises an infant. The intestinal tract of an unborn is, despite general belief, not sterile, but contains bacteria that have been transferred from the mother. This opens a new research field and may change our understanding about the role bacteria play in early life, the selection of strains with probiotic properties and the treatment of diseases related to bacterial infections. Differences in bacterial populations isolated from meconia may provide answers to the prevention of certain forms of diabetes. More research is now focusing on the effect that a genetically diverse group, versus a much simpler microbial population, may have on the development of a homeostatic gut microbiome. The effect different bacterial species have on the gut-associated lymphoid tissue and cascade of immune responses has been well researched, but we still fail in identifying the ideal group of intestinal bacteria and if we do, it will certainly not be possible to maintain homeostasis with so many challenges the gut faces. Changes in diet, antibiotics, food preservatives and stress are some of the factors we would like to control, but more than often fail to do so. The physiology and genetics of the GIT changes with age and so the microbiome. This review summarises factors involved in the regulation of a gut microbiome.

Safety assessment of antibiotic and probiotic feed additives for Gallus gallus domesticus.

Antibiotics in feed select for resistant strains and is thus a threat to human health. In this study, the effect of a multi-strain probiotic and antibiotics on the growth and health of broilers was studied. Equal numbers of broilers received on a daily basis either a multi-strain probiotic or a combination of sulphadiazine, colistin and trimethoprim, whereas the control group received standard feed. The villi of immature broilers (19 days old) administered antibiotics had a larger surface area and their lymphocyte and basophil counts were higher compared to broilers from the probiotic and control groups. The cecal microbiomes of mature broilers (29 days old) that received probiotics had higher levels of Enterobacteriaceae, but lower numbers of Clostridiales, Brucellaceae, Synergistaceae, Erysipelotrichaceae and Coriobacteriaceae compared to the antibiotic-treated group. A decline in the bioluminescence of Listeria monocytogenes observed for broilers on probiotics suggested that the probiotic may be used to control bacterial infections. No significant differences in total red blood cell, haemoglobin and haematocrit content, and mean values for corpuscular volume, corpuscular haemoglobin and corpuscular haemoglobin numbers were recorded amongst broilers from the different treatment groups. This study provides valuable information on the health and performance of broilers when administered probiotics and antibiotics as additives.

Surfactin-loaded polyvinyl alcohol (PVA) nanofibers alters adhesion of Listeria monocytogenes to polystyrene.

Surfactin-loaded polyvinyl alcohol (PVA) nanofibers were spun using gravity electrospinning. Scanning electron microscopy (SEM) images showed that nanofibers spun with surfactin are free from bead formation and uniform in diameter. The average nanofiber diameters were decreased (273±39nm, 259±39nm and 217±33nm) with increasing levels of surfactin (0.5, 1.0 and 1.5%, w/v) into PVA (10%, w/v). The 10% (w/v) PVA had average fiber diameter of 303±33nm. Atomic force microscopy (AFM) analysis showed that fibers spun with surfactin are not smooth as PVA fibers. The surface average roughness (Sa) estimated for surfactin loaded nanofibers (0.5%: 19.0nm, 1.0%: 20.4nm and 1.5%: 20.7nm) was higher as compared with PVA (10%:15.8nm). Scanning transmission electron microscopy (STEM) showed no matrix differences between PVA and surfactin-loaded PVA nanofibers. Fourier transform infrared (FTIR) microscopy revealed uniform distribution of surfactin in PVA. Based on differential scanning calorimetry (DSC) analyses, surfactin decreased the crystallinity of PVA during spinning. No antimicrobial activity was detected against methicillin-resistant Staphylococcus aureus (MRSA) strain Xen 30, Listeria monocytogenes EDGe, Escherichia coli Xen 14, and Pseudomonas aeruginosa PA01. However, the adhesion of L. monocytogenes to polystyrene in presence of surfactin-loaded nanofibers decreased significantly (OD595: 0.012±0.001) as compared with control (OD595: 0.022±0.002), suggesting that these nanofibers may be used in wound dressings or in the coating of prosthetic devices to prevent biofilm formation and secondary infections.

Polyethylene oxide (PEO)-hyaluronic acid (HA) nanofibers with kanamycin inhibits the growth of Listeria monocytogenes.

Listeria monocytogenes is well known to cause prosthetic joint infections in immunocompromised patients. In this study, polyethylene oxide (PEO) nanofibers, containing kanamycin and hyaluronic acid (HA), were prepared by electrospinning at a constant electric field of 10kV. PEO nanofibers spun with 0.2% (w/v) HA and 1% (w/v) kanamycin had a smooth, bead-free structure at 30-35% relative humidity. The average diameter of the nanofibers was 83±20nm. Attenuated total reflectance (ATR)-Fourier transform infrared (FTIR) spectroscopy indicated that kanamycin was successfully incorporated into PEO/HA matrix. The presence of kanamycin affects the thermal properties of PEO/HA nanofibers, as shown by differential scanning calorimetry (DSC) and thermogravimetric analyses (TGA). The kanamycin-PEO-HA nanofibers (1mg; 47±3µg kanamycin) inhibited the growth of L. monocytogenes EDGe by 62%, as compared with PEO-HA nanofibers, suggesting that it may be used to coat prosthetic implants to prevent secondary infections.

First Report of the Isolation of the Symbiotic Bacterium Photorhabdus luminescens subsp. laumondii Associated with Heterorhabditis safricana from South Africa.

Photorhabdus luminescens subsp. laumondii is closely associated with the entomopathogenic nematode Heterorhabditis bacteriophora and has, to date, not been isolated from other nematode species. This study is the first report of P. luminescens subsp. laumondii from two South African isolates of entomopathogenic nematodes, Heterorhabditis safricana SF281 and H. bacteriophora SF351. Both symbiotic bacterial strains are phenotypically closely related to P. luminescens subsp. laumondii previously isolated and described from H. bacteriophora. The genetic relatedness between P. luminescens subsp. laumondii strains SF281B and SF351B was confirmed by comparing 16S rDNA, recA, gyrB and gltX sequences with sequences of P. luminescens subsp. laumondii, including the type strain (TT01T) and strain E21.

Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 excludes Listeria monocytogenes from the GIT, as shown by bioluminescent studies in mice.

Listeria monocytogenes is an opportunistic food-borne pathogen and is life-threatening to individuals with a weakened immune system. The aim of this study was to determine if Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA could prevent colonisation of L. monocytogenes in the gastro-intestinal tract (GIT). Mice were gavaged with L. plantarum 423, E. mundtii ST4SA, and a combination of the two strains, for 6 consecutive days and orally infected with a bioluminescent strain of L. monocytogenes (strain EGDe) on the last day of treatment. 30 min after infection, high cell numbers of L. plantarum 423, E. mundtii ST4SA and L. monocytogenes EGDe were isolated from faeces. L. monocytogenes EGDe cells were absent from the small intestine of L. plantarum 423-treated mice 4 h after infection and from the large intestine 2 h later. No bioluminescent, and thus metabolically active, cells of L. monocytogenes EGDe were recorded in the GIT of mice treated with E. mundtii ST4SA, suggesting that their growth was repressed. L. plantarum 423 and E. mundtii ST4SA colonised the colon the strongest. These strains may be considered for the competitive exclusion of L. monocytogenes from the GIT.

First report of the symbiotic bacterium Xenorhabdus indica associated with the entomopathogenic nematode Steinernema yirgalemense.

The entomopathogenic nematode Steinernema yirgalemense is considered a promising agent in the biocontrol of insects. However, little is known about the bacteria living in symbiosis with the nematode. In this study, we have identified the only available bacterial strain (157-C) isolated from S. yirgalemense, as a member of the species Xenorhabdus indica. Identification was based on 16S rDNA, recA, dnaN, gltX, gyrB and infB gene sequence analyses. The relatedness of strain 157-C to the type strain of X. indica (DSM 17 382) was confirmed with DNA-DNA hybridization. The phenotypic characteristics of strain 157-C are similar to those described for the type strain of X. indica. This is the first report associating X. indica with S. yirgalemense.

Malting of barley with combinations of Lactobacillus plantarum, Aspergillus niger, Trichoderma reesei, Rhizopus oligosporus and Geotrichum candidum to enhance malt quality.

Good quality malt is characterised by the presence of high levels of fermentable sugars, amino acids and vitamins. To reach the starch-rich endosperm of the kernel, ß-glucan- and arabinoxylan-rich cell walls have to be degraded. ß-Glucanase is synthesized in vast quantities by the aleurone layer and scutellum during germination. Secretion of hydrolytic enzymes is often stimulated by addition of the plant hormone gibberellic acid (GA3) during germination. We have shown an enhanced ß-glucanase and α-amylase activity in malt when germinating barley was inoculated with a combination of Lactobacillus plantarum B.S1.6 and spores of Aspergillus niger MH1, Rhizopus oligosporus MH2 or Trichoderma reesei MH3, and L. plantarum B.S1.6 combined with cell-free culture supernatants from each of these fungi. Highest malt ß-glucanase activity (414 Units/kg malt) was recorded with a combination of L. plantarum B.S1.6 and spores of A. niger MH1. Highest α-amylase activities were recorded with a combination of L. plantarum B.S1.6 and spores of R. oligosporus MH2 (373 Ceralpha Units/g malt). Highest FAN levels were recorded when L. plantarum was inoculated in combination with spores of either R. oligosporus MH2 or T. reesei MH3 (259 and 260 ppm, respectively). This is the first study showing that cell-free culture supernatants of Aspergillus, Rhizopus and Trichoderma have a stimulating effect on ß-glucanase and α-amylase production during malting. A combination of L. plantarum B.S1.6, and spores of A. niger MH1 and R. oligosporus MH2 may be used as starter cultures to enhance malt quality.

The Effects of Continuous In Vivo Administration of Nisin on Staphylococcus aureus Infection and Immune Response in Mice.

Mice were intraperitoneally infected with 2 × 10(8) cfu Staphylococcus aureus Xen 36 and treated with 2,130 AU (arbitrary units) nisin (equivalent to 27.7 µg pure nisin), a class Ia lantibiotic, over 7 days. The metabolic activity of S. aureus Xen 36, concluded from changes in cell bioluminescence, declined for the first 3.5 h, but increased over the next 24 h and remained at this level for the remainder of the 7-day trial. Similar results were obtained with heat-inactivated (25 min at 121 °C) nisin, suggesting that the decline in metabolic activity of S. aureus Xen 36 cannot be attributed to the bacteriostatic activity of nisin. The decline in lymphocyte numbers in infected mice was of smaller magnitude after treatment with active nisin compared to inactive nisin, suggesting that active nisin limited the apoptosis of lymphocytes. The drastic increase in neutrophil versus lymphocyte (N:L) ratio observed in the presence of active nisin suggested that the decline in metabolic activity of S. aureus Xen 36 was due to an immune response triggered by the infection. Nisin, active or inactive, stimulated the activity of cytokines interleukin-6, interleukin-10 and tumour necrosis factor. However, the overall immune response triggered by both forms of nisin was too minute to trigger an abnormally high antigenic immune reaction.

Nisin F-loaded brushite bone cement prevented the growth of Staphylococcus aureus in vivo.

AIMS: To determine if nisin F-loaded self-setting brushite cement could control the growth of Staphylococcus aureus in vivo. METHODS AND RESULTS: Brushite cement was prepared by mixing equimolar concentrations of ß-tricalcium phosphate and monocalcium phosphate monohydrate. Nisin F was added at 5·0, 2·5 and 1·0% (w/w) and the cement moulded into cylinders. In vitro antibacterial activity was determined using a delayed agar diffusion assay. Release of nisin F from the cement was determined using BCA protein assays. Based on scanning electron microscopy and X-ray diffraction analysis, nisin F did not cause significant changes in cement structure or chemistry. Cement containing 5·0% (w/w) nisin F yielded the most promising in vitro results. Nisin F-loaded cement was implanted into a subcutaneous pocket on the back of mice and then infected with S. aureus Xen 36. Infection was monitored for 7 days, using an in vivo imaging system. Nisin F prevented S. aureus infection for 7 days and no viable cells were isolated from the implants. CONCLUSIONS: Nisin F-loaded brushite cement successfully prevented in vivo growth of S. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: Nisin F incorporated into bone cement may be used to control S. aureus infection in vivo.

Nisin F, intraperitoneally injected, may have a stabilizing effect on the bacterial population in the gastro-intestinal tract, as determined in a preliminary study with mice as model.

AIMS: To determine if nisin F has an effect on the bacterial population in the gastro-intestinal tract. METHODS AND RESULTS: Six male C57BL/6 mice were intraperitoneally injected with 200 µl sterile saline and six with nisin F (200 µl, equivalent to 640 arbitrary units). Fecal samples were collected before injection and 8, 24 and 48 h after injection, and the bacteria amplified by PCR-DGGE using 16S rDNA primers. The composition of the bacterial population in the gastro-intestinal tract (GIT) of mice that were injected with saline changed during 48 h, whereas the bacterial population in the GIT remained relatively unchanged in animals injected with nisin F. CONCLUSIONS: These results suggest that nisin F inhibits the growth of specific bacteria in the GIT within the first 4 h. Furthermore, the species remained repressed for at least 44 h after one intraperitoneal injection with nisin F. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report suggesting that nisin F may have a stabilizing effect on the bacterial population in the gastro-intestinal tract.

Influence of carbohydrates on the isolation of lactic acid bacteria.

AIMS: To determine the influence of carbohydrates on enrichment isolation of lactic acid bacteria from different niches. METHODS AND RESULTS: Lactic acid bacteria in three traditional fermented products in southern Africa (amasi, mahewu and tshwala) and in three fresh samples (two flowers and a fruit) were enrichment cultured in media supplemented with 13 different carbohydrates. Diversity of lactic acid bacteria was determined by PCR-denaturing-gradient gel electrophoresis. Carbohydrates used in enrichment media had a big impact on the isolation of lactic acid bacteria from fermented products. Depending on the carbohydrates tested, the number of species detected ranged from one to four in amasi, one to five in mahewu and one to three in tshwala. Fructose and mannitol selected for relatively higher numbers of lactic acid bacteria in fermented products. Specific relationships between substrates and lactic acid bacteria have been noted. On the other hand, small influences were found among carbohydrates tested in flowers and fruit. CONCLUSION: Carbohydrates have a big impact on the isolation of a variety of lactic acid bacteria in fermented food. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that reports the influence of carbohydrates on the enrichment of lactic acid bacteria.

The ability of nisin F to control Staphylococcus aureus infection in the peritoneal cavity, as studied in mice.

AIMS: To determine the ability of nisin F to control systematic infection caused by Staphylococcus aureus, using C57BL/6 mice as a model. METHODS AND RESULTS: Twelve mice were intraperitoneally injected with 1 × 10(8) viable cells of Staph. aureus Xen 36 containing the modified Photorhabdus luminescence luxABCDE operon on plasmid pAUL-A Tn4001. After 4 h, six mice were intraperitoneally injected with 640 arbitrary units (AU) nisin F, and six were injected with sterile saline. Six mice, not infected with Staph. aureus, were treated with nisin F, and six not infected were left untreated. The viability of Staph. aureus Xen 36 was monitored over 48 h by recording photon emission levels. Nisin F suppressed Staph. aureus for 15 min in vivo. No abnormalities were recorded in blood analyses and internal organs of mice treated with nisin F. CONCLUSIONS: Nisin F suppressed the growth of Staph. aureus in the peritoneal cavity for at least 15 min. Re-emergence of Staph. aureus bioluminescence over the next 44 h suggests that nisin F was inactivated, most probably by proteolytic enzymes. SIGNIFICANCE AND IMPACT OF THE STUDY: A single dosage of nisin F administered in the peritoneal cavity controlled the growth of Staph. aureus for at least 15 min in vivo.