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At least 19 recordsLinked to original sources
PubMed2004

Genetic characterization of non-spoilage variant isolated from beer-spoilage Lactobacillus brevis ABBC45.

AIMS: To characterize the non-spoilage variant obtained from beer-spoilage Lactobacillus brevis ABBC45C and to identify a potential genetic marker capable of discriminating beer-spoilage L. brevis strains from non-spoilers. METHODS AND RESULTS: A non-spoilage variant was obtained from beer-spoilage L. brevis ABBC45C by repeatedly subculturing the strain at 37 degrees C. Genetic characterization of the variant revealed that 12,605 bp portion of one plasmid, designated pRH45II, was lost in the variant. The sequence analysis indicates the presence of 12 ORFs in the deleted region of pRH45II. The PCR and Southern hybridization study revealed that the homologues of ORF5 found in the deleted region were present in all of the beer-spoilage L. brevis strains examined in this study. In contrast, the homlogues appeared to be absent in non-spoilage L. brevis strains. CONCLUSIONS: The presence or absence of ORF5 homologues was found to be highly correlated with the beer-spoilage ability of L. brevis strains, indicating this ORF is potentially a useful genetic marker capable of differentiating beer-spoilage strains among L. brevis. SIGNIFICANCE AND IMPACT OF THE STUDY: A non-spoilage variant was successfully isolated from beer-spoilage L. brevis ABBC45C. This study could facilitate the understanding of mechanisms underlying beer-spoilage ability of L. brevis.

Bacterial Proteins↗
PubMed2002

Food spoilage--interactions between food spoilage bacteria.

Food spoilage is a complex process and excessive amounts of foods are lost due to microbial spoilage even with modern day preservation techniques. Despite the heterogeneity in raw materials and processing conditions, the microflora that develops during storage and in spoiling foods can be predicted based on knowledge of the origin of the food, the substrate base and a few central preservation parameters such as temperature, atmosphere, a(w) and pH. Based on such knowledge, more detailed sensory, chemical and microbiological analysis can be carried out on the individual products to determine the actual specific spoilage organism. Whilst the chemical and physical parameters are the main determining factors for selection of spoilage microorganisms, a level of refinement may be found in some products in which the interactive behavior of microorganisms may contribute to their growth and/or spoilage activity. This review gives three such examples. We describe the competitive advantage of Pseudomonas spp. due to the production of iron-chelating siderophores, the generation of substrates for spoilage reactions by one organism from another microorganism (so-called metabiosis) and the up-regulation of phenotypes potentially involved in spoilage through cell-to-cell communication. In particular, we report for the first time the widespread occurrence of N-acyl homoserine lactones (AHL) in stored and spoiling fresh foods and we discuss the potential implications for spoilage and food preservation.

Bacteria↗
PubMed1964

BACTERIOLOGY OF SPOILAGE OF FISH MUSCLE. II. INCIDENCE OF SPOILERS DURING SPOILAGE.

A test medium consisting of a sterile raw press juice from fish muscle was used to determine the incidence of spoilage bacteria on stored fillets of English sole (Parophrys vetulus). The initial load of spoilers was shown to be consistently below 10%. This percentage rose but slightly toward the middle of the spoilage runs, and actually declined when spoilage was most apparent both organoleptically and chemically. Further evidence implicating the Pseudomonas and Achromobacter groups in the spoilage of fresh fish is presented.

Achromobacter↗
PubMed2004

A rapid method for the evaluation of both extrinsic and intrinsic contamination and resulting spoilage of water-in-oil emulsions.

AIMS: To develop a method for studying the microbial spoilage of water-in-oil emulsions and to use this to investigate (i) the intrinsic stability of water-in-oil formulations and (ii) Pseudomonas aeruginosa SP1-induced spoilage of a proprietary emulsion. METHODS AND RESULTS: Aliquots of test emulsion were placed into wells of a microtitre plate and the opacity (492 nm) monitored at 120-min intervals over several hours. Cracking of the emulsion was associated with marked reductions in opacity. Rate and extent of change in O.D. could be used as indicators of spoilage. Spoilage of a laboratory emulsion formulation was investigated where microorganisms with demonstrated spoilage potential were incorporated either into the water phase prior to emulsification or where the proportion of contaminated water droplets was varied by dilution of contaminated emulsion with a sterile formulation. Results suggested that the route of introduction was a critical determinant of the probability of gross spoilage. Ps. aeruginosa SP1-induced spoilage of a proprietary formulation was found to be independent of growth in the formulation; rather it was attributed to the presence of a heat-labile extracellular spoilage-factor that was protease labile and possessed both lipase and polysorbate hydrolytic activity. Such spoilage potential was unique to one Ps. aeruginosa culture filtrate amongst five pseudomonads tested. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is both rapid and reproducible, enables evaluation of the effects of route of contamination upon emulsion spoilage and has potential application in formulation development for cosmetic, pharmaceutical and food products.

Colony Count, Microbial↗
PubMed2004

Genetic characterization and specific detection of beer-spoilage Lactobacillus sp. LA2 and related strains.

AIMS: Lactobacillus sp. LA2 (DSM15502) and related strains (LA2 group) possess strong beer-spoilage ability. The 16S rDNA sequence of LA2 strain is virtually indistinguishable from that of L. collinoides, generally considered to be nonbeer-spoilage bacteria. The aim of this study was to identify the genetic marker to distinguish between Lactobacillus sp. LA2 group and L. collinoides and to provide a rapid means of identifying beer-spoilage strains belonging to Lactobacillus sp. LA2 group. METHODS AND RESULTS: The 16-23S rDNA intergenic spacer (ITS) regions of Lactobacillus sp. LA2 and L. collinoides JCM1123T were sequenced to identify a genetic marker to distinguish between the two groups. As a result, 300 and 500 bp ITS regions of Lactobacillus sp. LA2 were found to be almost identical with those of L. collinoides JCM1123T. Sequence comparison analysis between Lactobacillus sp. LA2 and L. collinoides JCM1123T revealed that the two contiguously located nucleotides are absent in both ITS regions of Lactobacillus sp. LA2. Based on the sequence difference, we have designed specific PCR primers with a minor modification to the primer sequence that can differentiate between beer-spoilage Lactobacillus sp. LA2 group and nonbeer-spoilage L. collinoides. CONCLUSIONS: The PCR-based method has been developed to identify Lactobacillus sp. LA2 group, providing a rapid and sensitive means of determining the beer-spoilage ability of detected bacterial strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The substitution of one nucleotide, located at the third position to the 3'-end in the primer sequence, enhanced the specificity of the PCR method while retaining sufficient sensitivity. The nucleotide gap identified in this study appeared to serve as a useful genetic marker that can differentiate 12 beer-spoilage Lactobacillus sp. LA2 group strains from its close relatives that exhibit no beer-spoilage ability.

Bacterial Typing Techniques↗
PubMed2005

Random amplified polymorphic DNA-PCR based cloning of markers to identify the beer-spoilage strains of Lactobacillus brevis, Pediococcus damnosus, Lactobacillus collinoides and Lactobacillus coryniformis.

AIMS: Beer-spoilage ability of lactic acid bacteria such as Lactobacillus brevis is a strain-dependent phenomenon in which the mechanism has not yet been completely clarified. In order to systematically identify genes that contribute to beer-spoilage, large-scale random amplified polymorphic DNA (RAPD)-based cloning methods was carried out. METHODS AND RESULTS: A systematic RAPD polymerase chain reaction (PCR) analysis using 600 primers was performed on beer-spoilage and on nonspoilage strains of L. brevis. Among 600 primers, three were found to amplify a single locus highly specific to beer-spoilage strains. DNA sequencing of this locus revealed a three-part operon encoding a putative glycosyl transferase, membrane protein and teichoic acid glycosylation protein. PCR analysis of typical beer-spoilage lactic acid bacteria suggested that this locus is highly specific to beer-spoilage strains. CONCLUSION: The cloned markers are highly specific to identify the beer-spoilage strains not only in L. brevis but also in Pediococcus damnosus, Lactobacillus collinoides and Lactobacillus coryniformis. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper proves that RAPD-PCR is an efficient method for cloning the strain-specific genes from bacteria. The markers described here is one of the most useful tools to identify the beer-spoilage strains of lactic acid bacteria.

Beer↗
PubMed2006

horC confers beer-spoilage ability on hop-sensitive Lactobacillus brevis ABBC45cc.

AIMS: To determine whether horC confers beer-spoilage ability and to evaluate the validity of horC as a trans-species genetic marker for differentiating the beer-spoilage ability of lactic acid bacteria (LAB). METHODS AND RESULTS: Hop-sensitive Lactobacillus brevis ABBC45cc was transformed with an expression plasmid, pHYchorBC, containing putative multidrug resistance gene horC and its putative regulator horB, and the transformant was designated as ABBC45cc/pHYchorBC. As a control, ABBC45cc was transformed with pHYchorB that contains horB, and the transformed strain was designated as ABBC45cc/pHYchorB. As a result of beer-spoilage assay of these transformants, ABBC45cc/pHYchorBC exhibited beer-spoilage ability, whereas ABBC45cc/pHYchorB did not. Furthermore ABBC45cc/pHYchorBC showed higher hop resistance than ABBC45cc/pHYchorB, accounting for the differences in beer-spoilage ability observed between the two transformants. ABBC45cc/pHYchorBC also exhibited higher resistance to various structurally unrelated drugs, compared with ABBC45cc/pHYchorB. CONCLUSIONS: horC was shown to confer hop resistance and beer-spoilage ability on ABBC45cc by presumably encoding a multidrug transporter. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that horC plays an important role in hop resistance and beer-spoilage ability supports the validity of horC as a trans-species genetic marker for differentiating the beer-spoilage ability of LAB.

Bacterial Proteins↗
PubMed2005

Isolation of a hop-sensitive variant of Lactobacillus lindneri and identification of genetic markers for beer spoilage ability of lactic acid bacteria.

We have isolated a hop-sensitive variant of the beer spoilage bacterium Lactobacillus lindneri DSM 20692. The variant lost a plasmid carrying two contiguous open reading frames (ORF s) designated horB(L) and horC(L) that encode a putative regulator and multidrug transporter presumably belonging to the resistance-nodulation-cell division superfamily. The loss of hop resistance ability occurred with the loss of resistance to other drugs, including ethidium bromide, novobiocin, and cetyltrimethylammonium bromide. PCR and Southern blot analysis using 51 beer spoilage strains of various species of lactic acid bacteria (LAB) revealed that 49 strains possessed homologs of horB and horC. No false-positive results have been observed for nonspoilage LAB or frequently encountered brewery isolates. These features are superior to those of horA and ORF 5, previously reported genetic markers for determining the beer spoilage ability of LAB. It was further shown that the combined use of horB/horC and horA is able to detect all 51 beer spoilage strains examined in this study. Furthermore sequence comparison of horB and horC homologs identified in four different beer spoilage species indicates these homologs are 96.6 to 99.5% identical, which is not typical of distinct species. The wide and exclusive distribution of horB and horC homologs among beer spoilage LAB and their sequence identities suggest that the hop resistance ability of beer spoilage LAB has been acquired through horizontal gene transfer. These insights provide a foundation for applying trans-species genetic markers to differentiating beer spoilage LAB including previously unencountered species.

Anti-Bacterial Agents↗
PubMed1995

Qualitative and quantitative characterization of spoilage bacteria from packed fish.

The large cells recently suggested to be responsible for spoilage of packed cod, have been identified as Photobacterium phosphoreum. The spoilage activity of these cells, of Shewanella putrefaciens and of other microorganisms isolated form spoiled packed cod has been studied. Both qualitative and quantitative tests were used for characterization of the microbial spoilage activity. The importance of the different groups of microorganisms was evaluated by comparison of microbial spoilage activity determined in model substrates and in product experiments. The yield factor for production of trimethylamine (YTMA/CFU) and the cell concentration determined at the time of off-odour detection were used as quantitative measurements of microbial spoilage activity. On average cells of P. phosphoreum produced 30 times more TMA than cells of S. putrefaciens, YTMA/CFU of the two organisms were 10(-8.0) mg-N TMA/cfu and 10(-9.5) mg-N TMA/cfu, respectively. With these yield factors the level of TMA found in spoiled packed cod (30 mg-N TMA/100g) corresponds to about 10(7) cfu/g of P. phosphoreum and to 10(8)-10(9) cfu/g of S. putrefaciens. 10(7) cfu/g of P. phosphoreum were actually found in spoiled packed cod suggesting this organism could be responsible for spoilage. High cell concentrations of more than 10(8) cfu/g of S. putrefaciens were required for production of detectable off-odours and is was concluded that this organism is without importance for spoilage of packed cod.

Animals↗
PubMed1996

Microbiological spoilage of fish and fish products.

Spoilage of fresh and lightly preserved fish products is caused by microbial action. This paper reviews the current knowledge in terms of the microbiology of fish and fish products with particular emphasis on identification of specific spoilage bacteria and the qualitative and quantitative biochemical indicators of spoilage. Shewanella putrefaciens and Pseudomonas spp. are the specific spoilage bacteria of iced fresh fish regardless of the origin of the fish. Modified atmosphere stored marine fish from temperate waters are spoiled by the CO2 resistant Photobacterium phosphoreum whereas Gram-positive bacteria are likely spoilers of CO2 packed fish from fresh or tropical waters. Fish products with high salt contents may spoil due to growth of halophilic bacteria (salted fish) or growth of anaerobic bacteria and yeasts (barrel salted fish). Whilst the spoilage of fresh and highly salted fish is well understood, much less is known about spoilage of lightly preserved fish products. It is concluded that the spoilage is probably caused by lactic acid bacteria, certain psychotrophic Enterobacteriaceae and/or Photobacterium phosphoreum. However, more work is needed in this area.

Animals↗
PubMed1996

Microbial and biochemical spoilage of foods: an overview.

During harvesting, processing and handling operations food may become contaminated with a wide range of microorganisms. Subsequently, during distribution and storage only a small fraction of these will develop and cause serious deteriorations. Which microorganisms will develop or what (bio)chemical reactions occur is dependent upon food derived or environmental factors. This paper will describe the main mechanisms involved in the loss of food quality for the most important food commodities. Food spoilage may be caused by a wide range of reactions including some that are mainly physical or chemical, others due to action of enzymes or microorganisms. The primary factors associated with food spoilage are associated with intrinsic food properties (e.g., endogenous enzymes, substrates, sensitivity for light, oxygen) and (cross)contamination during harvesting, slaughter and processing in combination with temperature abuse. For fresh foods the primary quality changes may be categorized as (i) bacterial growth and metabolism resulting in possible pH-changes and formation of toxic compounds, off-odours, gas and slime-formation, (ii) oxidation of lipids and pigments in fat-containing foods resulting in undesirable flavours, formation of compounds with adverse biological effects or discoloration. Although interrelated with the microbial spoilage, the last category is 'purely' chemical in nature and will, all other things being equal, increase in importance with decreasing temperature. Little is known about the relationship between microbial activity and (bio)chemical spoilage parameters under different packaging and storage conditions. Although there is much progress in the characterisation of the total microflora and metabolites developing during spoilage, not much is known about the identification of specific microorganisms in relation to food composition. Despite the fact that food spoilage is a huge economical problem world wide, it is obvious that the mechanisms and interaction leading to food spoilage are very poorly understood.

Color↗
PubMed1996

Specific spoilage organisms in breweries and laboratory media for their detection.

The Gram positive bacteria are generally regarded as the most hazardous beer spoilage organisms in modern breweries, especially the lactobacilli: L. brevis, L. lindneri, L. curvatus, L. casei, L. buchneri, L. coryneformis, L. plantarum, L. brevisimilis, L. malefermentans and L. parabuchneri and the pediococci: P damnosus, P. inopinatus and P. dextrinicus. Micrococcus kristinae is the only species within the micrococci relevant to brewing. The Gram negative strictly anaerobic bacteria are apparently increasing in importance and include Pectinatus cerevisiiphilus, Pectinatus frisingensis and Selenomonas lacticifex, reported as obligate beer spoilage organisms: Zymophilus raffinosivorans as a potential beer spoilage organism; Megasphaera cerevisiae as an obligate spoilage organism of low alcohol beer and Zymomonas mobilis as capable of spoiling primed beer. With improved process technology the importance of aerobic bacteria has decreased and the same applies for the Gram negative aerobic bacteria Hafnia protea and Enterobacter cloacae which are capable of surviving beer fermentation. Beer spoilage organisms include several so-called wild yeasts, of which Saccharomyces species are generally considered the most important. Even though the detection of beer spoilage organisms by cultivation in laboratory media does not always provide the specificity and the sensitivity required, the use of selective media and incubation conditions still appears to be the method preferred by breweries. The media used depend on the type of sample, the specificity required and, for detection of wild yeasts, to some extent, the characteristics of the culture yeast. Among the media reported so far no single medium can be used to detect all members within a group of specific beer spoilage organisms and further work on the development of improved substrates are required both for bacteria and wild yeasts.

Bacteria↗
PubMed2006

Effect of bacterial interactions on the spoilage of cold-smoked salmon.

Cold-smoked salmon is a lightly preserved fish product in which a mixed microbial flora develops during storage and where the interactive behaviour of micro-organisms may contribute to their growth and spoilage activity. The aim of this study was to assess the effect of the bacterial interactions between the main species contaminating the cold-smoked salmon on bacterial growth, chemical and sensory changes, and spoilage. First, Carnobacterium piscicola, Photobacterium phosphoreum, Lactobacillus sakei, Vibrio sp., Brochothrix thermosphacta and Serratia liquefaciens-like were inoculated as pure cultures on sterile cold-smoked salmon. All bacterial species grew well; Vibrio sp. was the fastest and L. sakei strains developed very rapidly as well with a high maximum cell density on cold-smoked salmon blocks (up to 10(9) cfu g(-1) after 10 days at 8 degrees C). Based on sensory analysis, Vibrio sp. was identified as non-spoilage bacteria, C. piscicola as very lightly and B. thermosphacta as lightly spoiling. L. sakei and S. liquefaciens-like were found to be the most spoiling bacteria. Secondly, C. piscicola and L. sakei, two species frequently occurring in the lactic flora of the product, were inoculated together and each of them in mixed cultures with respectively P. phosphoreum, Vibrio sp., B. thermosphacta, and S. liquefaciens-like. The growth of L. sakei was shown to strongly inhibit most of the co-inoculated strains i.e. P. phosphoreum, B. thermosphacta, S. liquefaciens-like and, to a lesser extent, Vibrio sp. The growth of C. piscicola seemed to be enhanced with B. thermosphacta and to develop earlier with P. phosphoreum and Vibrio sp. Conversely, S. liquefaciens-like and P. phosphoreum were weakly inhibited by C. piscicola. The main observation resulting from the sensory evaluation was the delay in the appearance of the spoilage characteristics in the mixed cultures with L. sakei, in particular L. sakei/ S. liquefaciens-like. On the other hand, the spoilage activity of the non-spoiler strains Vibrio sp. or the moderate spoilage strains B. thermosphacta and C. piscicola was increased when they were associated together. It is concluded that the spoilage behaviour of micro-organisms in mixed culture is significantly different from pure culture and explain the difficulty to find robust quality indices for this product.

Animals↗
PubMed2003

Identification of lactic acid bacteria from spoilage associations of cooked and brined shrimps stored under modified atmosphere between 0 degrees C and 25 degrees C.

AIMS: To evaluate spoilage and identify lactic acid bacteria (LAB) from spoilage associations of cooked and brined shrimps stored under modified atmosphere packaging (MAP) at 0, 5, 8, 15 and 25 degrees C. METHODS AND RESULTS: Bacterial isolates (102) from spoilage associations of cooked and brined MAP shrimps were characterized by phenotypic tests and identified as lactic acid bacteria (78 isolates), other Gram-positive bacteria (13 isolates) and Gram-negative bacteria (11 isolates). A selection of 48 LAB isolates were further characterized and identified by phenotypic tests and SDS-PAGE electrophoresis of whole cell proteins. Selected clusters of LAB isolates were analysed by plasmid profiling, pulsed field gel electrophoresis and 16S rRNA gene sequencing. Enterococcus faecalis was identified in spoilage associations at 15 degrees C and 25 degrees C, and its metabolic activity corresponded to chemical changes in spoiled products. Carnobacterium divergens, a non-motile Carnobacterium sp. nov. and Lactobacillus curvatus were the LAB species observed in spoilage associations of products stored at 0 degrees C, 5 degrees C and 8 degrees C. CONCLUSIONS: Enterococcus spp. and Carnobacterium spp. were the dominant parts of spoilage associations of cooked and brined MAP shrimps stored at high and low temperatures, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The SDS-PAGE technique and simple biochemical keys allowed the majority of LAB isolates from spoilage associations of cooked and brined MAP shrimps to be identified at the species level.

Animals↗
PubMed2005

Post harvest spoilage of sweetpotato in tropics and control measures.

Sweetpotato storage roots are subjected to several forms of post harvest spoilage in the tropical climate during transportation from farmers' field to market and in storage. These are due to mechanical injury, weight loss, sprouting, and pests and diseases. Sweetpotato weevil is the single most important storage pest in tropical regions for which no control measures or resistant variety are yet available. Several microorganisms (mostly fungi) have been found to induce spoilage in stored sweetpotatoes. The most important among them are Botryodiplodia theobromae, Ceratocystis fimbriata, Fusarium spp., and Rhizopus oryzae. The other less frequently occurring spoilage microorganisms include Cochliobolus lunatus (Curvularia lunata), Macrophomina phaseolina, Sclerotium rolfsii, Rhizoctonia solani, Plenodomus destruens. Microbial spoilage of sweetpotato is found associated with decrease in starch, total sugar, organic acid (ascorbic acid and oxalic acid) contents with concomitant increase in polyphenols, ethylene, and in some instances phytoalexins. Several methods are used to control microbial spoilage. Curing to promote wound healing is found as the most suitable method to control microbial spoilage. Curing naturally occurs in tropical climates where mean day temperature during sweetpotato harvesting season (February-April) invariably remains at 32-35 degrees C and relative humidity at 80-95%. Sweetpotato varieties varied in their root dry matter content, and low root dry matter content attributed for their high curing efficiency. Curing efficiency of varieties also differed in response to curing periods. Fungicide treatment, bio-control, gamma irradiation, hydro warming, and storage in sand and saw dust were found to have intermediate impacts in controlling spoilage and enhancing shelf life of sweetpotato roots. Breeding program has to be chalked out to develop new varieties suitable to curing under tropical conditions in addition to developing varieties having multi-spectrum resistance to major post harvest rot pathogens and sweetpotato weevils.

Food Contamination↗
PubMed2004

Presence of acylated homoserine lactones (AHLs) and AHL-producing bacteria in meat and potential role of AHL in spoilage of meat.

Quorum-sensing (QS) signals (N-acyl homoserine lactones [AHLs]) were extracted and detected from five commercially produced vacuum-packed meat samples. Ninety-six AHL-producing bacteria were isolated, and 92 were identified as Enterobacteriaceae. Hafnia alvei was the most commonly identified AHL-producing bacterium. Thin-layer chromatographic profiles of supernatants from six H. alvei isolates and of extracts from spoiling meat revealed that the major AHL species had an R(f) value and shape similar to N-3-oxo-hexanoyl homoserine lactone (OHHL). Liquid chromatography-mass spectrometry (MS) (high-resolution MS) analysis confirmed the presence of OHHL in pure cultures of H. alvei. Vacuum-packed meat spoiled at the same rate when inoculated with the H. alvei wild type compared to a corresponding AHL-lacking mutant. Addition of specific QS inhibitors to the AHL-producing H. alvei inoculated in meat or to naturally contaminated meat did not influence the spoilage of vacuum-packed meat. An extracellular protein of approximately 20 kDa produced by the H. alvei wild-type was not produced by the AHL-negative mutant but was restored in the mutant when complemented by OHHL, thus indicating that AHLs do have a regulatory role in H. alvei. Coinoculation of H. alvei wild-type with an AHL-deficient Serratia proteamaculans B5a, in which protease secretion is QS regulated, caused spoilage of liquid milk. By contrast, coinoculation of AHL-negative strains of H. alvei and S. proteamaculans B5a did not cause spoilage. In conclusion, AHL and AHL-producing bacteria are present in vacuum-packed meat during storage and spoilage, but AHL does not appear to influence the spoilage of this particular type of conserved meat. Our data indicate that AHL-producing H. alvei may induce food quality-relevant phenotypes in other bacterial species in the same environment. H. alvei may thus influence spoilage of food products in which Enterobacteriaceae participate in the spoilage process.

4-Butyrolactone↗