A numerical taxonomic study of lactic acid bacteria from vacuum-packed beef, pork, lamb and bacon

S haw B. G. & H arding C harmaigne D. 1984. A numerical taxonomic study of lactic acid bacteria from vacuum packed beef, pork, lamb and bacon. Journal of Applied Bacteriology 56 , 25–40.
A numerical taxonomic study using 79 unit characters has been performed on 100 isolates of lactic acid bacteria from refrigerated vacuum-packed beef, pork, lamb and bacon. Three clusters were observed at 78% S which contained all the strains apart from three unidentifiable streptobacteria, one Leuconostoc , and one strain of Pediococcus pentosaceus . One cluster (III) consisted of only one strain of Leuc. paramesenteroides and six unidentifiable Leuconostoc strains. The two largest clusters (I and II) were both composed entirely of streptobacteria. Cluster I contained 31 strains (G + C content 33–2–36–9 moles %) which were not identifiable with any described species. Cluster II contained 57 strains (G + C content 40–7–43–7 moles %) which were provisionally identified with Lactobacillus sake or Lact. bavaricus according to the lactic acid isomer produced. The division of nearly all the streptobacteria into two clearly defined clusters has resolved problems which have existed in the classification of lactic acid bacteria from vacuum-packed meat.     

Classification of some lactic acid bacteria from vacuum-packed meats by direct probe mass spectrometry

The technique of direct probe mass spectrometry (DPMS) has been applied to the classification of 40 strains of lactic acid bacteria from refrigerated vacuum-packed beef, pork, lamb and bacon. Relationships between strains were examined by multi-variate statistical techniques using sets of ions selected for reproducibility and sample discrimination. Five groups were distinguished which corresponded closely to those detected in a previous numerical taxonomic study. Two groups contained all 12 representatives of a cluster of unidentifiable non-aciduric streptobacteria whose sub-division is supported by other taxonomic evidence. All twenty-one strains from a cluster of aciduric streptobacteria provisionally identified with Lacto-bacillus sake were contained in two further groups. The sub-division of these acid-uric strains revealed by DPMS has not been verified by other techniques and requires further investigation. The fifth group contained Leuconostoc strains. The study demonstrates the value of DPMS in confirming and clarifying classification schemes obtained by conventional methods.     

Classification of some lactic acid bacteria from vacuum-packed meats by direct probe mass spectrometry

The technique of direct probe mass spectrometry (DPMS) has been applied to the classification of 40 strains of lactic acid bacteria from refrigerated vacuum-packed beef, pork, lamb and bacon. Relationships between strains were examined by multi-variate statistical techniques using sets of ions selected for reproducibility and sample discrimination. Five groups were distinguished which corresponded closely to those detected in a previous numerical taxonomic study. Two groups contained all 12 representatives of a cluster of unidentifiable non-aciduric streptobacteria whose sub-division is supported by other taxonomic evidence. All twenty-one strains from a cluster of aciduric streptobacteria provisionally identified with Lactobacillus sake were contained in two further groups. The sub-division of these aciduric strains revealed by DPMS has not been verified by other techniques and requires further investigation. The fifth group contained Leuconostoc strains. The study demonstrates the value of DPMS in confirming and clarifying classification schemes obtained by conventional methods.     

Diversity of enterococcal bacteriocins and their grouping in a new classification scheme

Enterococci are lactic acid bacteria of importance in food, public health and medical microbiology. Many strains produce bacteriocins, some of which have been well characterized. This review describes the structural and genetic characteristics of enterocins, the bacteriocins produced by enterococci. Some of these can be grouped with typical bacteriocins produced by lactic acid bacteria according to traditional classification, whereas others are atypical and structurally distinct from the general classes of bacteriocins. These atypical enterocins recently played an important role in and prompted reclassification of the class II bacteriocins into a new scheme. In this review, a more simplified classification scheme for enterocins based on amino acid sequence homologies is proposed. Enterocins are of interest for their diversity and potential for use as food biopreservatives. The emergence of multiple antibiotic-resistant enterococci among agents of nosocomial disease and the presence of virulence factors among food isolates requires a careful safety evaluation of isolates intended for potential biotechnical use. Nevertheless, enterococcal bacteriocins produced by heterologous hosts or added as cell-free preparations may still be attractive for application in food preservation.     

Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential

Two traditional fermented food 'tapai' (fermented tapioca) and 'tempoyak' (fermented durian flesh), chilli puree and fresh goat's milk were used as sources for the isolation of lactic acid bacteria (LAB). A total of 126 isolates were obtained and by sequential screening for catalase activity and Gram-staining, 55 were determined to be LAB out of which 16 were established to be homofermentative by the gel plug test. Seven isolates were identified by use of the API 50CHL kit and two lactobacilli strains and one lactococci strain were selected to study their growth and lactic acid production profiles in a time course experiment. The lactobacilli strains, both isolated from 'tapai', produced higher amounts of cells and lactic acid from glucose as compared to the lactococci strain isolated from fresh goat's milk.     

A novel enteropathogenic, strongly haemolytic spirochaete isolated from pig and mallard, provisionally designated 'Brachyspira suanatina' sp. nov

Atypical, strongly haemolytic porcine isolates of intestinal spirochaetes differing genetically from Brachyspira hyodysenteriae were identified and characterized. The isolates were subjected to culture and biochemical tests, antimicrobial susceptibility testing and molecular analyses. None of four species-specific polymerase chain reaction systems targeting genes of B. hyodysenteriae gave a positive reaction. All the atypical porcine isolates were identical in their partial 16S rRNA and nox gene sequences with a previously described isolate from a mallard (Anas platyrhynchos), and differed only slightly from another mallard isolate. All these isolates were distinctly different from all currently recognized Brachyspira species. A challenge study was carried out using recently weaned pigs. Clinical signs and macroscopic changes consistent with swine dysentery were seen both in pigs given the atypical porcine isolate and in control pigs given the reference strain of B. hyodysenteriae (B204(R)). Pigs given the genetically similar isolate from a mallard became colonized and diarrhoea was observed. This is the first study indicating that Brachyspira isolates from mallard can infect pigs and induce diarrhoea. We propose that this atypical spirochaete genotype should be regarded as a new species within the genus Brachyspira, and be provisionally designated 'Brachyspira suanatina' sp. nov.     

Characterisation of lactic acid producing <Emphasis Type="Italic">Sporolactobacillus</Emphasis> strains from tree barks in Thailand

Eight spore-forming lactic acid producing bacteria were isolated from tree barks in Thailand. They were identified as Sporolactobacillus nakayamae (Group I, three isolates), S. terrae (Group II, two isolates), S. kofuensis (Group III, one isolate) and S. inulinus (Group IV, two isolates) based on their phenotypic characteristics and 16S rRNA gene sequence analyses. Four isolates in Groups I and II produced DL lactic acid (89.60–114.61 g/L), while three isolates in Groups III and IV produced D-lactic acid (88.01–113.78 g/L). Isolate BK65-3 identified as S. inulinus produced the highest D-lactic acid concentrations (101.42 g/L), productivity (1.41 g/L/h), yields (84.52%) and optical purity of D-lactic acid (100%).     

Isolation and identification of lactic acid bacteria from soil using an enrichment procedure

AIMS: To survey, and identify and classify the ecological distribution of lactic acid bacteria from soil in Japan and Taiwan. METHODS AND RESULTS: Acid-producing bacteria were isolated from 68 soil samples, collected from Japan and Taiwan, in the rhizospheres of fruit trees, from the floor of a henhouse and around a horse farm. All isolates were identified by physiological and genetic tests. Thirty-two of the 54 isolates were identified as lactic acid bacteria (LAB), 16 as spore-forming lactic acid bacteria, five as Clostridium and one as Bacillus. These lactic acid bacteria represent five genera: Lactobacillus, Lactococcus, Enterococcus, Leuconostoc and Weissella. CONCLUSIONS: A high rate of isolating lactic acid bacteria was obtained from soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that soil may be a common source for the isolation of lactic acid bacteria.     

Assessment of aflatoxin and cyclopiazonic acid production by Aspergillus flavus isolates from Hungary

Thirty-two isolates of Aspergillus flavus were obtained from various sources in Hungary. All isolates were morphologically identified as A. flavus and three atypical variants were confirmed as A. flavus by comparing their DNA with an ex type culture of A. flavus. None of these isolates produced aflatoxins when tested on coconut agar or grown on rice medium and culture extracts examined by thin layer chromatography. Also, none of the isolates converted sterigmatocystin, O-methyl sterigmatocystin, norsolorinic acid, or sodium acetate to aflatoxin. However, 59% of the isolates produced cyclopiazonic acid based on thin layer chromatographic analysis of culture extracts. The isolates that lack the ability to produce both aflatoxin and cyclopiazonic acid are potential candidates for use in bicontrol studies.     

Production of histamine and tyramine by lactic acid bacteria isolated from vacuum-packed sugar-salted fish

The incidence of histamine- or tyramine-producing lactic acid bacteria was examined in several products of vacuum-packed sugar-salted fish (salmon, halibut, mackerel). No histamine-producing isolates were observed, whereas the majority of tyramine-producing isolates were identified as Carnobacterium spp. These organisms were shown to be important members of the microbial flora during storage of vacuum-packed sugar-salted salmon at 5°C. The amount of tyramine produced was reduced by lowering the temperature from 9°C to 4°C for all of five strains of carnobacteria or lactobacilli. The majority of tyramine was produced during the exponential growth phase for Carnobacterium piscicola N 5 and Lactobacillus viridescens N 69. The ability of these bacteria to produce tyramine may be used as an index of microbial quality/acceptability of stored vacuum-packed sugar-salted fish.     

Isolation and characterization of fructophilic lactic acid bacteria from fructose-rich niches

Fourteen strains of fructophilic lactic acid bacteria were isolated from fructose-rich niches, flowers, and fruits. Phylogenetic analysis and BLAST analysis of 16S rDNA sequences identified six strains as Lactobacillus kunkeei, four as Fructobacillus pseudoficulneus, and one as Fructobacillus fructosus. The remaining three strains grouped within the Lactobacillus buchneri phylogenetic subcluster, but shared low sequence similarities to other known Lactobacillus spp. The fructophilic strains fermented only a few carbohydrates and fermented d-fructose faster than d-glucose. Based on the growth characteristics, the 14 isolates were divided into two groups. Strains in the first group containing L. kunkeei, F. fructosus, and F. pseudoficulneus grew well on d-fructose and on d-glucose with pyruvate or oxygen as external electron acceptors, but poorly on d-glucose without the electron acceptors. Strains in this group were classified as “obligately” fructophilic lactic acid bacteria. The second group contained three unidentified strains of Lactobacillus that grew well on d-fructose and on d-glucose with the electron acceptors. These strains grew on d-glucose without the electron acceptors, but at a delayed rate. Strains in this group were classified as facultatively fructophilic lactic acid bacteria. All fructophilic isolates were heterofermentative lactic acid bacteria, but “obligately” fructophilic lactic acid bacteria mainly produced lactic acid and acetic acid and very little ethanol from d-glucose. Facultatively fructophilic strains produced lactic acid, acetic acid and ethanol, but at a ratio different from that recorded for heterofermentative lactic acid bacteria. These unique characteristics may have been obtained through adaptation to the habitat.     

Ultrasound imaging in an experimental model of fatty liver disease and cirrhosis in rats

Background

Atypical scrapie was first identified in Norwegian sheep in 1998 and has subsequently been identified in many countries. Retrospective studies have identified cases predating the initial identification of this form of scrapie, and epidemiological studies have indicated that it does not conform to the behaviour of an infectious disease, giving rise to the hypothesis that it represents spontaneous disease. However, atypical scrapie isolates have been shown to be infectious experimentally, through intracerebral inoculation in transgenic mice and sheep. The first successful challenge of a sheep with 'field' atypical scrapie from an homologous donor sheep was reported in 2007.

Results

This study demonstrates that atypical scrapie has distinct clinical, pathological and biochemical characteristics which are maintained on transmission and sub-passage, and which are distinct from other strains of transmissible spongiform encephalopathies in the same host genotype.

Conclusions

Atypical scrapie is consistently transmissible within AHQ homozygous sheep, and the disease phenotype is preserved on sub-passage.     

Assessment of genetic variability and relatedness among atypical Aeromonas salmonicida from marine fishes,using AFLP-fingerprinting

Atypical strains of Aeromonas salmonicida are the causal agent of atypical furunculosis or ulcer disease in various fish species, including spotted wolffish Anarhichas minor, which is a promising species in the Norwegian fish-farming industry. Isolates of atypical A. salmonicida comprise a very heterogenous group showing large variety in biochemical, molecular and virulence characteristics. The genetic variability among atypical isolates from wolffish was characterised using amplified fragment length polymorphism analysis: AFLP-fingerprinting. Additional isolates from halibut Hippoglossus hippoglossus, turbot Scophthalmus maximus, cod Gadus morhua and several salmonid fishes were included for assessment of variability and relatedness among a total of 56 atypical isolates of A. salmonicida. They were compared to reference strains of A. salmonicida subspecies and to other Aeromonas species pathogenic in fishes. AFLP-fingerprints subjected to similarity analysis yielded a grouping of the isolates into several clusters, revealing genetic heterogeneity among the isolates. There seems to be a correlation between genetic similarity among isolates and the fish host. The Icelandic isolates, mainly from cod, formed a very homogeneous subcluster, which was closely related to the wolffish isolates. All atypical isolates from spotted and common wolffish grouped together in a large cluster and appear to be very homogeneous, even though they had been isolated over a period of 8 yr at different locations in Norway. On the other hand, most of the isolates from turbot and halibut grouped together into 2 different clusters, while the 9 atypical isolates from salmonids appeared in 4 different clusters. Thus, the atypical isolates of A. salmonicida from halibut, turbot and salmonid fishes seem to be more genetically diverse than those from wolffish and cod.     

The occurrence and properties of Gemmiger formicilis and related anaerobic budding bacteria in the avian caecum

Gram negative budding bacteria with a characteristic morphology have been observed in the caeca of chickens, turkeys, ducks and guinea fowl at levels between 0–4 and 24% of the total flora. Using strict anaerobic techniques a detailed study has been made of the organisms isolated from the caeca of chickens aged between 15 and 44 d. The majority of isolates were identified as Gemmiger formicilis , the strains being divided into twomajor types according to the relative amounts of butyric acid and lactic acid produced from glucose. A second group of isolates could not be related to any known species.
Amongst properties of ecological interest studied were limiting temperatures for growth, ammonia utilization, ability to utilize various carbon sources and survival in environments outside the intestine.
Attempts to produce a selective medium for the isolation of these organisms when present at less than 2% of the total flora were unsuccessful.     

Identification ofAcetobacter strains isolated from spoiled lactic acid fermented meat food for pets

FiveAcetobacter isolates from lactic acid fermented meat food for pets were characterized by 177 morphological, physiological and biochemical traits. Four isolates were identified asA. pasteurianus, one asA. aceti. It is emphasized that access of such bacteria to lactic acid fermented foods should be avoided.     

The fermentation of cell wall substances in grass silage and in potato pulp

Summary The amount of acid formed in grass silage was greater than could have been formed from the soluble sugars present, even when only a lactic fermentation took place. This seemed to point to fermentation of cell wall substances by lactic acid bacteria. Lactic acid fermentation in potato pulp always takes place with cell wall substances as substrates, as sugars are absent. It was found that galactose, probably occurring as galactan, and also some pectic acid were fermented in potato pulp. Some lactobacilli were isolated from potato pulp; streptobacteria which could ferment galactan but no pectic or galacturonic acid, and betabacteria which could ferment galacturonic acid but no galactan or pectic acid. A number of homofermentative lactobacilli were all found to belong to the speciesStreptobacterium casei. It was shown that a strain of this species could ferment galactan in potato pulp sterilised previously with ethylene oxide. Part of this work was carried out at the Netherlands Institute for Dairy Research, Ede, Netherlands.     

Isolation and characterization of lactic acid bacteria from yan-taozih (pickled peaches) in Taiwan

Two hundred cultures of lactic acid bacteria were isolated from eight yan-taozih (pickled peaches) samples, and four cultures were isolated from the main component of these samples, fresh peaches. Phenotypic and biochemical characteristics identified eight different bacterial groups (A–H) and showed that the majority of the isolates was heterofermentative lactic acid bacteria. The most common bacterial species in yan-taozih was Leuconostoc mesenteroides, although regional diversity was also observed among the lactic acid bacteria strains isolated. The antibacterial activities of the isolates were also determined, and 20 isolates were found to show inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157^T. The results of the sensory evaluation suggested that the diversity of lactic acid bacteria has a great effect on the aroma and formation of taste in the food product. Our findings suggest that Leuconostoc mesenteroides is the most abundant lactic acid bacteria in yan-taozih (pickled peaches) and that lactic acid bacteria strains play an important role in affecting the aroma and taste of the food product.     

One-tube oxidation-fermentation methods: limitations posed by atypical fermentative reactions.

Atypical fermentation in the two-tube method of Hugh and Leifson (1953), whereby acid was produced in the sealed tube but not in the open tube, was determined to be the result of a preferential metabolism of peptone over the reactant carbohydrate. These atypical fermentations are only partially detected by one-tube oxidation-fermentation methods, and as such they constitute a source of limitation to the routine use of these abbreviated techniques for oxidative-fermentation determinations.     

新疆发酵乳品中乳酸菌的分离鉴定及其耐药性

目的对新疆传统发酵乳品中乳酸菌进行分离鉴定并检测其耐药性。方法利用传统形态学鉴定法和生化鉴定等方法对新疆发酵乳中乳酸菌进行鉴定,采用纸片扩散法对分离鉴定的菌进行耐药性分析。结果从新疆发酵乳品中共分离出8株乳酸菌,经鉴定分别为瑞士乳杆菌(Lactobacillus helveticus)、植物乳杆菌(Lactobacillus plantarum)、马乳酒样乳杆菌(Lactobacillus kefianofaciens)、乳酸乳球菌(Lactococcus lactis)、副干酪乳杆菌(Lactobacillus paracasei)、副干酪乳杆菌类坚韧亚种(Lactobacillus paracasei subsp.tolerans)、哈尔滨乳杆菌(Lactobacillus harbinensis)、希氏乳杆菌(Lactobacillus hilgardii),并且发现8株乳酸菌对万古霉素、庆大霉素、阿莫西林、多西环素、环丙沙星、阿奇霉素、头孢他啶、头孢孟多具有一定敏感性。结论新疆发酵乳品中以乳杆菌居多,对常见抗生素具有一定的敏感性。     

Widespread distribution of the bacteriocin-producing lactic acid cocci in Miso-paste products

AIMS: A survey was conducted on the ecological distribution of bacteriocin-producing lactic acid bacteria in Miso-pastes, a typical fermented food in Japan. METHODS AND RESULTS Nine Miso-pastes were sampled for isolation of bacteriocin-producers. Almost all isolated enterococcal strains produced bacteriocins but no isolated tetragenococci did so. The bacteriocin-producing isolates were divided into nine groups by phenotypic tests. As the phenotypic characters were highly diverse, these strains could not be identified to species level on the basis of their phenotypes. The nine representative strains from each group were identified by 16S rRNA analysis. These bacteriocin-producers with one exception (Lactococcus sp.) were identified as strains of the Enterococcus faecium 'species group'. The bacteriocins of the nine strains were classified into five types according to their antibacterial spectral patterns and their SDS-PAGE profiles. The bacteriocins inhibited undesirable bacteria in Miso-pastes, such as Bacillus subtilis, but did not inhibit the useful Tetragenococcus halophila. CONCLUSIONS: The bacteriocin-producing lactic acid cocci were widespread at high frequencies in Miso-pastes. They were considered to play an important role in preventing the growth of undesirable bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that bacteriocin-producers act effectively as safe biopreservatives and may contribute to the biopreservation in Miso-pastes.