Bacteriological quality and risk assessment of the imported and domestic bottled mineral water sold in Fiji.

Considering the popularity of bottled mineral water among indigenous Fijians and tourists alike, a study was carried out to determine the bacteriological quality of different bottled waters. A risk assessment was also carried out. Seventy-five samples of bottled mineral water belonging to three domestic brands and 25 samples of one imported brand were analysed for heterotrophic plate count (HPC) bacteria and faecal coliforms. HPC counts were determined at 22 degrees C and 37 degrees C using R2A medium and a membrane filtration technique was used to determine the faecal coliform (FC) load in 100 ml of water on mFC agar. Between 28 and 68% of the samples of the various domestic brands failed to meet the WHO standard of 100 colony forming units (cfu) per 100 ml at 22 degrees C and 7% of these also tested positive for faecal coliforms. All imported bottled mineral water samples were within WHO standards. A risk assessment of the HPC bacteria was carried out in terms of beta haemolytic activity and antibiotic resistance. More than 50% of the isolates showed beta haemolytic activity and were multi-drug resistant. While the overall quality of the product was generally good, there is a need to enforce stringent quality standards for the domestic bottlers to ensure the safety of consumers.

Microbiological effect of mupirocin and chlorhexidine for Staphylococcus aureus decolonization in community and nursing home based adults.

OBJECTIVE: To compare the presence of Staphylococcus aureus and pathogenic Gram-negative rods (GNR) in the anterior nares, posterior pharynx and three skin sites in community-based adults and nursing home-based adults before and after treatment with nasal mupirocin and topical chlorhexidine. METHODS: S. aureus-colonized adults were recruited from the community (n=26) and from nursing homes (n=8). Eligible participants were cultured for S. aureus and GNR during two study visits and then received intranasal mupirocin and topical chlorhexidine for 5days, with a 2-month follow-up period. RESULTS: After decolonization, we found sustained decreases of S. aureus colonization in nose, throat and skin sites over 4-8weeks in both populations. Intranasal mupirocin did not increase GNR colonization in nose or throat. Chlorhexidine did not decrease GNR colonization in skin sites. CONCLUSIONS: Decolonization with mupirocin and chlorhexidine leads to a sustained effect on S. aureus colonization without affecting GNR colonization.

Purification and antipathogenic activity of lipid transfer proteins (LTPs) from the leaves of Arabidopsis and spinach.

Two homogeneous proteins active in vitro against the bacterial pathogen Clavibacter michiganensis subsp. sepedonicus were obtained from a crude cell-wall preparation from the leaves of Columbia wild-type Arabidopsis. The N-terminal amino acid sequences of these proteins allowed their identification as lipid transfer proteins (LTP-a1, LTP-a2); the LTP1-a1 sequence was identical to that deduced from a previously described cDNA (EMBL M80566) and LTP-a2 was quite divergent (44% identical positions). These proteins were not detected in the cytoplasmic fraction by Western-blot analysis. Proteins LTP-s1 and LTP-s2 were similarly obtained from spinach leaves; LTP-s1 was 91% identical to a previously purified spinach LTP (Swiss Prot P10976), and LTP-s2 was moderately divergent (71% identical positions). About 1/3 of the total LTPs were detected in the cytoplasmic fraction from spinach by Western-blot analysis. Concentrations of these proteins causing 50% inhibition (EC-50) were in the 0.1-1 microM range for the bacterial pathogens C. michiganensis and Pseudomonas solanacearum and close to 10 microM for the fungal pathogen Fusarium solani.

Aureobacterium resistens sp. nov., exhibiting vancomycin resistance and teicoplanin susceptibility.

Two similar strains of a coryneform bacterium were isolated from human clinical material. Both strains were resistant to vancomycin but susceptible to teicoplanin. Detailed biochemical, chemotaxonomical, and molecular genetic investigations revealed that both isolates were members of a hitherto undescribed species of genus Aureobacterium. The name Aureobacterium resistens sp. nov. is proposed for the new bacterium and the type strain is CCUG 38312.

Effects of environmental factors on leucine catabolism by Carnobacterium piscicola.

The metabolites from leucine degradation are involved in dry fermented sausage aroma. The catabolism of leucine by a strain of Carnobacterium piscicola was studied directly in the growth medium with 3H-labelled leucine to investigate the effect of five parameters: phase of growth, pH, oxygen, glucose and alpha-ketoisocaproic acid. Resting cells (RC) were also incubated with 3H-labelled leucine. The radioactive metabolites from leucine catabolism were analysed by high performance liquid chromatography (HPLC). At pH 5.4 and 7.2, the main metabolites detected were 3-methyl butanal, 3-methyl butanol and alpha-ketoisocaproic acid. At pH 6.5, the leucine catabolism was maximum and was characterised by a high production of 3-methyl butanoic acid. Leucine catabolism was most important during the exponential phase of growth. The addition of alpha-ketoisocaproic acid at 1%, glucose at levels of 0.5% to 2% and shaking of the growth medium increased leucine catabolism.

Antimicrobial effect of natural preservatives in a cooked and acidified chicken meat model.

The inhibitory effect of Microgard 100, Microgard 300, nisin, Alta 2002, Perlac 1902, sodium lactate and essential oil of mustard on microorganisms experimentally inoculated was screened in an acidified chicken meat model (pH = 5.0) and stored for 2 weeks at a none restrictive growth temperature of 22 degrees C. All antimicrobials tested were used at the highest concentration recommended by their manufacturer. Sausage batter made with mechanically deboned chicken was inoculated with a mixed culture of Escherichia coli ATCC 25922, Brochothrix thermosphacta CRDAV452, and a protective culture Lactobacillus alimentarius BJ33 (FloraCan L-2). A final cell concentration of 3-4 log CFU g (-1) was targeted after cooking at a core temperature of 55 degrees C for each microorganism in order to assess cell count variation effectively. Composition, water activity (a(w)), pH and redox potential of the sausage model was also evaluated. The E. coli population decreased steadily during storage and was close or below detection level (< 1 log CFU g (-1)) for all treatments, including the control, after 14 days. Sodium lactate was most effective against B. thermosphacta; population was 4 log lower than the control after 14 days of storage. When essential oil of mustard was used, aerobic mesophilic bacteria and lactic acid bacteria were significantly lower than the control after 2 days of storage (P < or = 0.05). The other antimicrobial agents tested had no significant effect on the aerobic mesophilic bacteria, E. coli, B. thermosphacta and lactic acid bacteria counts, when compared to the control.

Interaction of monolaurin, eugenol and sodium citrate on growth of common meat spoilage and pathogenic organisms.

Interactions of monolaurin, eugenol (phenolic compound) and sodium citrate (chelator) on the growth of six organisms including common meat spoilage (Lactobacillus curvatus, Lactobacillus sake, Leuconostoc mesenteroides, Brochothrix thermosphacta) and pathogenic (Escherichia coli O157:H7 and Listeria monocytogenes) organisms were investigated. The combinations of 100 to 250 ppm monolaurin with 500 and 1000 ppm eugenol, and 0.2 and 0.4% sodium citrate were more effective than each component separately. More than one combination prevented detectable growth of each organism. Lactic acid bacteria (LAB) and E. coli O157:H7 were most resistant and L. monocytogenes and B. thermosphacta most sensitive to control by the chosen combinations. The presence of sodium citrate was necessary to yield potent inhibition of Lb. curvatus and Lb. sake growth by the monolaurin and eugenol combinations.

An improved protocol for electroporation of Oenococcus oeni ATCC BAA-1163 using ethanol as immediate membrane fluidizing agent.

AIMS: To finalize an effective and reproducible electroporation procedure to transform Oenococcus oeni ATCC BAA-1163 strain. METHODS AND RESULTS: The vector pGID052 was selected to optimize the electroporation procedure. Transformation efficiency was 5.8 x 10(3) per microg of DNA. Transformation was improved when competent cells were prepared with exponential phase cultures; optimum electroporation parameters were an electric pulse of 125 kV cm(-1), under a resistance of 200 omega and the presence of 10% (v/v) ethanol in the electroporation buffer (EPB). CONCLUSIONS: An effective protocol to transform O. oeni ATCC BAA-1163 strain by electroporation has been obtained by addition of ethanol to the EPB. A heterologous expression was obtained in O. oeni ATCC BAA-1163 by introducing a recombinant vector encoding a truncated form of ClpL2 protein. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a successful electroporation of O. oeni ATCC BAA-1163. The major improvement was the addition of ethanol to the EPB, which has never been reported before as means of enhancing the incorporation of foreign DNA molecules into prokaryote cells by electroporation. This method constitutes a useful tool for the genetic study of this lactic bacterium.

Novel chemolithotrophic, thermophilic, anaerobic bacteria Thermolithobacter ferrireducens gen. nov., sp. nov. and Thermolithobacter carboxydivorans sp. nov.

Three thermophilic strains of chemolithoautotrophic Fe(III)-reducers were isolated from mixed sediment and water samples (JW/KA-1 and JW/KA-2(T): Calcite Spring, Yellowstone N.P., WY, USA; JW/JH-Fiji-2: Savusavu, Vanu Levu, Fiji). All were Gram stain positive rods (approximately 0.5 x 1.8 microm). Cells occurred singly or in V-shaped pairs, and they formed long chains in complex media. All utilized H(2) to reduce amorphous iron (III) oxide/hydroxide to magnetite at temperatures from 50 to 75 degrees C (opt. approximately 73 degrees C). Growth occurred within the pH(60C) range of 6.5-8.5 (opt. pH(60C) 7.1-7.3). Magnetite production by resting cells occurred at pH(60C) 5.5-10.3 (opt. 7.3). The iron (III) reduction rate was 1.3 mumol Fe(II) produced x h(-1) x ml(-1) in a culture with 3 x 10(7) cells, one of the highest rates reported. In the presence or absence of H(2), JW/KA-2(T) did not utilize CO. The G + C content of the genomic DNA of the type strain is 52.7 +/- 0.3 mol%. Strains JW/KA-1 and JW/KA-2(T) each contain two different 16S rRNA gene sequences. The 16S rRNA gene sequences from JW/KA-1, JW/KA-2(T), or JW/JH-Fiji-2 possessed >99% similarity to each other but also 99% similarity to the 16S rRNA gene sequence from the anaerobic, thermophilic, hydrogenogenic CO-oxidizing bacterium 'Carboxydothermus restrictus' R1. DNA-DNA hybridization between strain JW/KA-2(T) and strain R1(T) yielded 35% similarity. Physiological characteristics and the 16S rRNA gene sequence analysis indicated that the strains represent two novel species and are placed into the novel genus Thermolithobacter within the phylum 'Firmicutes'. In addition, the levels of 16S rRNA gene sequence similarity between the lineage containing the Thermolithobacter and well-established members of the three existing classes of the 'Firmicutes' is less than 85%. Therefore, Thermolithobacter is proposed to constitute the first genus within a novel class of the 'Firmicutes', Thermolithobacteria. The Fe(III)-reducing Thermolithobacter ferrireducens gen. nov., sp. nov. is designated as the type species with strain JW/KA-2(T) (ATCC 700985(T), DSM 13639(T)) as its type strain. Strain R1(T) is the type strain for the hydrogenogenic, CO-oxidizing Thermolithobacter carboxydivorans sp. nov. (DSM 7242(T), VKM 2359(T)).

Septicemia due to Solobacterium moorei in a patient with multiple myeloma.

We report a case of bacteremia caused by Solobacterium moorei, an anaerobic, non-sporulated Gram-positive bacillus in a patient with a multiple myeloma. The source of infection was presumably related to multiple dento-alveolar abscesses. This is the first recovery of S. moorei from blood cultures.

Endophthalmitis due to Microbacterium species: case report and review of microbacterium infections.

Microbacterium species (formerly CDC [Centers for Disease Control and Prevention] coryneform group A-4 and A-5 bacteria) are widely distributed in the environment and rarely cause infections in humans. We present a case of endophthalmitis due to Microbacterium species that occurred after accidental trauma and review the literature on microbacterium infections. If the infected tissue or medical device is removed and antimicrobial therapy (preferably with beta-lactams or glycopeptides) is instituted, the prognosis is usually favorable for patients with microbacterium infections.

Reduction of Brochothrix thermosphacta on beef surfaces following immobilization of nisin in calcium alginate gels.

Lean and adipose beef carcass tissues inoculated with Brochothrix thermosphacta (BT) (approx. 4.50 log10 cfu cm-2) were left untreated (U) or treated with 100 micrograms ml-1 nisin (N), calcium alginate (A) or 100 micrograms ml-1 nisin immobilized in a calcium alginate gel (AN). Tissue samples were refrigerated after treatments and bacterial populations and nisin activity were determined at 0, 1, 2 and 7 d. U, A and N treatments of lean and adipose tissues did not suppress bacterial growth ( > 6 log10 cfu cm-2 by day 7) while treatments of lean and adipose tissues with AN suppressed bacteria ( > 2.42 log10 cfu cm-2 by day 7). Bacteriocin titres from both tissues were higher in AN vs N samples after the 7 d incubation. This study demonstrates that immobilization of nisin in a gel may be a more effective delivery system of a bacteriocin to the carcass surface than direct application.

Isolation and properties of a bacteriocin-producing Carnobacterium piscicola isolated from fish.

A facultative psychrotrophic lactic acid bacterium isolated from fresh fish was identified as Carnobacterium piscicola on the basis of carbohydrate utilization, G + C content and 16S RNA analysis. Its bacteriocin, designated carnocin UI49, is produced during the mid-exponential phase of growth at temperatures between 15 degrees C and 34 degrees C. Carnocin UI49 is active against a large number of closely-related lactic acid bacteria including carnobacteria, lactobacilli, pediococci and lactococci. Furthermore, the bacteriocin has a bactericidic mode of action which results in lysis of sensitive cells. Maximum bactericidal activity is observed at 34 degrees C with a decrease in activity down to 15 degrees C where it is completely abolished.

In vitro susceptibilities of aerobic and facultative non-spore-forming gram-positive bacilli to HMR 3647 (RU 66647) and 14 other antimicrobials.

The comparative in vitro activity of the ketolide HMR 3647 (RU 66647) and those of structurally related macrolide-lincosamide-streptogramin compounds (erythromycin, roxithromycin, azithromycin, clarithromycin, josamycin, lincomycin, pristinamycin, and quinupristin-dalfopristin) as well as those of benzylpenicillin, doxycycline, vancomycin, teicoplanin, levofloxacin, and rifapentine against 247 aerobic and facultative non-spore-forming gram-positive bacilli were determined by an agar dilution method. The ketolide was active against most organisms tested except Corynebacterium striatum, coryneform CDC group 12, and Oerskovia spp. The frequency of resistance to erythromycin and other macrolides as well as that to lincomycin was high. Pristinamycin and, to a lesser extent, quinupristin-dalfopristin were very active, but resistance to these agents was present in some strains of Rhodococcus equi, Listeria spp., C. striatum, Erysipelothrix rhusiopathiae, and Oerskovia spp. HMR 3647 was very active against all erythromycin-sensitive and many erythromycin-nonsusceptible strains, especially Corynebacterium minutissimum, Corynebacterium pseudodiphtheriticum, Corynebacterium amycolatum, and Corynebacterium jeikeium. In vitro resistance to benzylpenicillin was common, but doxycycline, vancomycin, and teicoplanin were very active against most organisms tested except E. rhusiopathiae, against which glycopeptide antibiotics were not active. The in vitro activity of levofloxacin was remarkable, but resistance to this agent was common for C. amycolatum, Corynebacterium urealyticum, C. jeikeium, and Oerskovia spp. strains. Rifapentine was also very active in vitro against many organisms, but resistance to this agent was always present in E. rhusiopathiae and was very common in C. striatum and C. urealyticum.

Incorporation of nisin into a meat binding system to inhibit bacteria on beef surfaces.

In two separate experiments, the bacteriocin, nisin, was incorporated into a commercially available meat binding system (Fibrimex) and applied to meat surfaces as a way of inhibiting the meat spoilage organism, Brochothrix thermosphacta during extended refrigerated storage. In experiment 1, pre-rigor lean beef carcass tissue (BCT) was inoculated with B. thermosphacta, left untreated (U), treated with 10 micrograms ml-1 nisin (N), Fibrimex (F) or Fibrimex containing 10 micrograms ml-1 nisin (FN), held aerobically at 4 degrees C for up to 7 d, and populations of B. thermosphacta and nisin activity determined. Experiment 2 determined the effects of the same treatments but on post-rigor, frozen and thawed lean BCT that was inoculated, vacuum-packaged, and stored at 4 degrees C for up to 14 d. In both experiments, N- and FN-treated tissues exhibited significantly lower populations of B. thermosphacta compared to U- and F-treated tissues, for the duration of refrigerated storage. Nisin activity was detected up to 7 d in N- and FN-treated samples from experiment 1. However, activity was detected only to days 0 and 2 in FN- and N-treated samples, respectively, from experiment 2. These studies indicate that the addition of a bacteriocin to a meat binding system and application to meat surfaces may be useful in reducing undesirable bacteria in restructured meat products.

Evaluation of stannous fluoride and chlorhexidine sprays on plaque and gingivitis in handicapped children.

The objective of this study was to evaluate the effect of twice-daily oral sprays of 2 ml chlorhexidine (0.2%) and 2 ml stannous fluoride (0.2%) as the sole oral hygiene measure on plaque and gingivitis in handicapped children. 52 institutionalized mentally handicapped individuals (aged 10-26 years) were divided into 4 groups to participate in a 9-week, double-blind, randomized clinically controlled trial, which included a cross-over. For the first 3 weeks, groups 3 and 2 had their mouths sprayed with chlorhexidine and stannous fluoride, respectively. Following a 3-week wash-out interval, groups 1 and 4 were sprayed with chlorhexidine and stannous fluoride, respectively. The alternate groups received a placebo, water. Plaque (PI) and gingival indices (GI) were recorded. Subgingival plaque samples were collected and counts of spirochaetes, motile rods and cocci were taken using darkfield microscopy. By the 9th week of trial, the PI and GI were reduced by 48% and 52% for the stannous fluoride group. In the chlorhexidine group, reductions of 75% (PI) and 78% (GI) were achieved. Pair-wise comparisons of placebo, stannous fluoride and chlorhexidine using the U-test of Mann-Whitney revealed significant (p less than 0.05) differences for both PI and GI scores in the placebo/stannous fluoride and placebo/chlorhexidine pair by the 9th week of the trial. In the stannous fluoride/chlorhexidine pair, only the PI was significantly reduced for those on chlorhexidine. Coccoid cells were more dominant at sites with low PI and GI scores (stannous fluoride and chlorhexidine groups), while spirochaetes and motile rods were more frequent at sites with high PI and GI scores (placebo group).

Vancomycin-resistant Aureobacterium species cellulitis and bacteremia in a patient with acute myelogenous leukemia.

A 39-year-old male with acute myelogenous leukemia and concomitant porphyria cutanea tarda was admitted to the hospital for consolidation chemotherapy of his leukemia. During his hospitalization, he developed cellulitis of the left hand and persistent bacteremia with a yellow-pigmented, nonfermenting coryneform bacterium that was identified as Aureobacterium sp. The portal of entry for the Aureobacterium infection was probably through the skin lesions due to porphyria cutanea tarda. The infection developed while the patient was receiving vancomycin prophylaxis, and the vancomycin MIC for the isolate was 32 micrograms/ml.

Improved antimicrobial activity of nisin-incorporated polymer films by formulation change and addition of food grade chelator.

AIMS: The following polymers were developed: polyethylene (PE), a PE and polyethylene oxide (70% PE and 30% PEO; PE + PEO) blend, PE and nisin (PE + nisin), PE, nisin, and EDTA (PE + nisin + EDTA), and PE + PEO with nisin (PE + PEO + nisin). METHODS AND RESULTS: Of the polymers tested, PE and PE + PEO did not exhibit any antimicrobial activity against Brochothrix thermosphacta (BT); however, PE + nisin, PE + nisin + EDTA, and PE + PEO + nisin did. Beef surfaces were experimentally inoculated with 3.50 log10 cfu/cm2 of BT, vacuum packaged with each of the five polymers, and held at 4 degrees C for 21 d. After 3 d at 4 degrees C, BT was reduced > 1.70 log(10) by PE + nisin and > 3.50 log(10) with PE + nisin + EDTA or PE + PEO + nisin. By 21 d at 4 degrees C, BT was reduced to 0.30 log(10) cfu/cm(2) when treated with PE + PEO + nisin. CONCLUSION: It appears that PE + PEO + nisin or PE + nisin + EDTA were more effective for reducing BT, as compared to polymers composed of PE + nisin. SIGNIFICANCE AND IMPACT OF THE STUDY: Nisin-incorporated polymers may control the growth of undesirable bacteria, thereby extending the shelf life and possibly enhancing the microbial safety of meats.