Microbiological Evaluation of Pacific Shrimp Processing

Microbiological evaluation of Pacific shrimp (Pandalus jordani) processing was made from samples obtained at five key processing points. The microbial count of raw shrimp ranged from 1.3 x 10(6) to 3.0 x 10(6). The initial microbial flora, in order of predominance, was Acinetobacter-Moraxella, Flavobacterium, Pseudomonas, gram-positive cocci, and Bacillus species. No yeasts were isolated. Differences in processing practices influenced both microbial count and the shrimp flora. The microbial load, however, always increased after peeling and sorting operations and decreased after cooking, washing, and brining steps. Significantly, the gram-positive cocci were recovered with increasing frequency after each processing step, reaching 76% of the total load in a final product. Most of them, however, were coagulase-negative.     

Foot odor due to microbial metabolism and its control

To characterize foot odor, we analyzed its components by sensory tests, isolated microorganisms that produce it, and evaluated the mechanism of the occurrence of foot odor. As a result, foot odor was found to be derived from isovaleric acid, which is produced when Staphylococcus epidermidis, a resident species of the normal cutaneous microbial flora, degrades leucine present in sweat. In addition, Bacillus subtilis was detected in the plantar skin of subjects with strong foot odor, and this species was shown to be closely associated with increased foot odor. Therefore, we screened various naturally occurring substances and fragrant agents that inhibit microbial production of foot odor without disturbing the normal microbial flora of the human skin. As a result, we identified citral, citronellal, and geraniol as fragrant agents that inhibit the generation of isovaleric acid at low concentrations.     

Molecular characterization of Lactobacillus curvatus and Lact. sake isolated from sauerkraut and their application in sausage fermentations

Lactobacillus curvatus and Lact. sake are best adapted to meat fermentations and dominate the flora during the whole process. In fermenting sauerkraut, Leuconostoc mesenteroides subsp. mesenteroides is the major organism only during the early phase. In this environment Lact. curvatus and Lact. sake provide up to 50% of the microbial flora especially of the later phase, depending on the process conditions. Strains of Lact. curvatus and Lact. sake isolated from fermenting sauerkraut were identified by hybridization with species specific 23S rRNA-targeted oligonucleotide probes and further characterized. In 59 of 72 strains, plasmid DNA was detected. Small cryptic plasmids of 20 strains were found to be homologous with pLc2, a 2·6 kb plasmid from Lact. curvatus LTH683, which was originally isolated from meat. The ability to compete was investigated in fermenting sausages of two strains each of Lact. curvatus and Lact. sake isolated from sauerkraut. One strain each of Lact. curvatus and Lact sake was found to outnumber the meat-borne flora and govern the process.     

In Vitro Communities Derived from Oral and Gut Microbial Floras Inhibit the Growth of Bacteria of Foreign Origins

The gastrointestinal (GI) tract is home to trillions of microbes. Within the same GI tract, substantial differences in the bacterial species that inhabit the oral cavity and intestinal tract have been noted. While the influence of host environments and nutritional availability in shaping different microbial communities is widely accepted, we hypothesize that the existing microbial flora also plays a role in selecting the bacterial species that are being integrated into the community. In this study, we used cultivable microbial communities isolated from different parts of the GI tract of mice (oral cavity and intestines) as a model system to examine this hypothesis. Microbes from these two areas were harvested and cultured using the same nutritional conditions, which led to two distinct microbial communities, each with about 20 different species as revealed by PCR-based denaturing gradient gel electrophoresis analysis. In vitro community competition assays showed that the two microbial floras exhibited antagonistic interactions toward each other. More interestingly, all the original isolates tested and their closely related species displayed striking community preferences: They persisted when introduced into the bacterial community of the same origin, while their viable count declined more than three orders of magnitude after 4 days of coincubation with the microbial flora of foreign origin. These results suggest that an existing microbial community might impose a selective pressure on incoming foreign bacterial species independent of host selection. The observed inter-flora interactions could contribute to the protective effect of established microbial communities against the integration of foreign bacteria to maintain the stability of the existing communities.     

Psychrotrophic bacterial flora of raw ewes' milk, with particular reference to gram negative rods

The microbial flora of 141 samples of raw ewes' milk was determined, before and after storage for 72 h at 4 degrees and 7 degrees C. Penicillin-resistant bacteria represented ca 61% of 1760 psychrotrophic isolates from refrigerated milk samples. Pseudomonas fluorescens and Pseudomonas fluorescent group-related strains predominated (ca 86%) in the Gram negative psychrotrophic microflora. Leuconostoc dextranicum was the most frequent Gram positive psychrotrophic species isolated.     

Comprehensive Analysis of Secondary Dental Root Canal Infections: A Combination of Culture and Culture-Independent Approaches Reveals New Insights

Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa), followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and –independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis.     

Psychrotrophic bacterial flora of raw ewes'milk, with particular reference to Gram negative rods

The microbial flora of 141 samples of raw ewes'milk was determined, before and after storage for 72 h at 4° and 7°C. Penicillin-resistant bacteria represented ca 61% of 1760 psychrotrophic isolates from refrigerated milk samples. Pseudomonas fluorescens and Pseudomonas fluorescent group-related strains predominated ( ca 86%) in the Gram negative psychrotrophic microflora. Leuconostoc dextranicum was the most frequent Gram positive psychrotrophic species isolated.     

A survey on the microbiological changes during the manufacture of dry-cured lacón, a Spanish traditional meat product

This article describes a microbiological study carried out on lacón, a dry-cured meat product made in the north-west of Spain from the fore extremity of pig. Using classical methods, aerobic mesophilic flora, salt-tolerant flora, lactic acid bacteria, Enterobacteriaceae, enterococci, moulds and yeasts were enumerated, some physicochemical parameters (pH, aw and moisture and NaCl contents) were determined and a representative number of isolates of the salt-tolerant flora (the main microbial group) were identified during the manufacture of five batches. All the microbial groups, with the exception of Enterobacteriaceae and enterococci, reached maximum counts both on the surface and in the interior of the pieces at the end of the post-salting stage and afterwards progressively dropped during the drying-ripening stage. Staphylococcus xylosus, Staph. saprophyticus, Staph. simulans, Staph. sciuri and Micrococcus luteus were the main species isolated throughout manufacturing. This study will significantly increase knowledge of the microbiology of cured meat products made from entire pieces.     

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora.     

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation.

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora.     

Characterization of micrococcaceae isolated from salt used for Spanish dry-cured ham

The microbial flora of salt used in the production of Spanish dry-cured ham was studied. The results indicated that Micrococcaceae constituted the predominant flora. Identification of the 369 isolates belonging to the family Micrococcaceae revealed that 60% belonged to genus Staphylococcus, 25% to Micrococcus, 6% to Kocuria, 5.4% to Dermacoccus and 0.5% to Stomatococcus. The species most often isolated was Staph. xylosus (28.9%), followed by M. lylae (21.4%), Staph. equorum (18.55%) and D. nishinomiyaensis (5.4%). The results indicate that the salt during salting process of dry-cured hams offers an ecosystem suitable for the survival of the staphylococci and micrococci.     

Histological and microbiological aspects of the vagina in captive howler monkeys (Alouatta caraya)

The histology and the glycogen content of the vaginal epithelium was studied in 15 adultAlouatta caraya monkeys. The histological sections revealed a thin epithelium without cornification. The application of the PAS-amylase method showed absence of glycogen. The microbial flora of the vagina was examined in eight adultAlouatta caraya. The vaginal smears showed a mixed flora with a predominance of Gram+cocci. Haemolytic streptococci were found in 100% of the tested monkeys. Other cocci, such asStaphylococcus albus, Streptococcus faecalis andMicrococcus sp. were less frequent. Of the fecal organisms,Proteus sp. was the most frequently isolated. Neither ureaplasms nor any species ofLactobacillus could be isolated. Positive cultures for nonpathogenic yeasts was a common feature. The relation between the absence of glycogen in the vaginal epithelium and the impossibility to isolate lactobacilli is discussed.     

Rare bacterium of new genus isolated with prolonged enrichment culture

Dynamic change in microbial flora was monitored with an oxygen electrode. The 1st phase microorganisms, which first grew well in LB medium, were followed by the 2nd phase microorganisms, which supposedly assimilated microbial cells of the 1st phase and their metabolites. In a similar way, a change in microbial flora was observed from the 1st phase to the 4th phase in 84 hr. Based on this observation, prolonged enrichment culture was done for as long as two months to increase the ratio of existence of rare microorganisms. From these culture liquids, four slow-growing bacteria (provisionally named Shinshu-ah1, -ah2, -ah3, and -ah4), which formed scarcely visible small colonies, were isolated. Sequence analysis of their 16S rDNA showed that Shinshu-ah1 had 97% homology with Bradyrhizobium japonicum and uncultured alpha proteobacterium clone blaii 16, Shinshu-ah2 91% with Rasbo bacterium, Alpha proteobacterium 34619, Bradyrhizobium genosp. P, Afipia felis and an unidentified bacterium, Shinshu-ah3 99% with Methylobacterium mesophilicum, and Shinshu-ah4 95% with Agromyces ramosus DSM 43045. Phylogenetic study indicated that Shinshu-ah2 had a possibility to form a new family, Shinshu-ah1 a new genus, and Shinshu-ah4 a new species.     

The gram-positive tonsillar and nasal flora of piglets before and after weaning

AIMS: To investigate gram-positive nasal and tonsillar microbial flora of piglets before and after weaning. METHODS AND RESULTS: The nasal and tonsillar gram-positive bacterial flora of 20 non-weaned piglets (2 weeks of age) and 20 weaned piglets (6 weeks of age), obtained from four different piggeries, was quantified by culture and identified by tDNA-PCR. The most widely occurring species from nasal conchae before as well as after weaning in the different piglets investigated were Streptococcus suis and Rothia nasimurium. After weaning a wide variety of Lactobacillus species appeared but in low numbers. In the tonsils, Strep. suis, Strep. dysgalactiae, S. hyicus, S. aureus, Arcanobacterium pyogenes and Actinomyces hyovaginalis were the species isolated from the largest number of pigs before and after weaning. S. aureus and most lactobacilli became more prevalent after weaning. Bacteria not known to be associated with pigs found in the present study included R. nasimurium, Strep. gallolyticus, Pediococcus pentosaceus and some Lactobacillus species. CONCLUSIONS: Over 30 different gram-positive bacterial species may occur in nasal conchae and tonsils of unweaned piglets at 2 weeks of age and of 6-week-old weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that weaning is associated with changes in prevalence of only a small minority of the highly diversified bacterial flora of the nares and tonsils of pigs.     

Intraspecific variation ofDrosophila buzzatii larval breeding success onOpuntia: A yeast-plant-insect relationship

The cactophilic species,Drosophila buzzatii, normally breeds in decaying pockets ofOpuntia cladodes, in which there is a complex interaction with the microbial flora, especially yeast species. Isofemale lines were used to estimate genetic variation among larvae reared on their natural feeding substrate. Four naturally occurring cactophilic yeast species isolated from the same Tunisian oasis as theDrosophila population were used. Two fitness components were studied for each line, viability and developmental time. Genetic variations amongD. buzzatii lines were observed for both traits. A significant yeast species x isofemale line interaction for viability was also evidenced, suggesting the occurrence of specialized genotypes for the utilization of breeding substrates. This genetic heterogeneity in the natural population may favor a better adaptation to the patchily distribution of yeasts.     

胃肠道微生态系统及其功能研究

随着生物技术和生理科学研究的不断深入,人们对胃肠道微生物生态及其功能的研究取得了重大进步。对胃肠道微生物的多样性、不同生理时期微生物菌群的演化和特定微生物种属的研究有新的发现。胃肠道微生物生态系统的生理功能得到不断的揭示,成为许多学者和多个学科关注的焦点。     

豆天蛾幼虫肠道细菌的初步研究

目的从微生态学角度探索豆天蛾的营养生理活动,了解其肠道菌群构成,找出具有利用价值的菌类,为豆天蛾的资源化开发和生物防治提供一定理论依据和技术基础。方法按照传统的培养分离方法从豆天蛾幼虫肠道消化道内分离纯化获得23个不同菌株,分别对其培养性状、菌体形态、染色反应和生理生化性状进行了系统研究。结果得到以葡萄球菌属（Staphylococcus）、地衣芽胞杆菌（B．lichenifomis）、丙酸杆菌属（Propionibacterium）为主的细菌23种,并记录了各种细菌的存在情况及数量。结论豆天蛾的菌群种类与其生活习性有关,为进一步研究适合豆天蛾幼虫生长和繁殖的人工饲料提供数据参考。     

Observations on the Microbiology of Cooked Chicken Carcasses

S ummary . The residual microbial flora and the flora developing during storage at 1–3° and at 16°, of chicken carcasses cooked in a circulating moist air oven operated at 85°, have been studied. All parts of the carcasses reached and maintained 85° for at least 50 min, and the residual flora consisted largely of spore forming bacteria. The predominant residual species were Bacillus subtilis and Clostridium bifermentans. Non-sporing bacteria were not detected after cooking nor after storage at 1–3° for up to 7 days. Storage at 16° for 3 days markedly increased the number of non-sporing organisms although off-odours typical of spoilage were not apparent until at least 10 days. Staphylococcus aureus and Salmonella spp. were not detected after cooking and storage and Cl. welchii was rarely isolated. It is concluded that poultry cooked by this method present a minimal risk of food-borne infection or intoxication by these organisms if contamination after cooking is avoided, the carcasses are cooled rapidly to c , 3° and stored at this temperature or frozen.     

Bacterial Populations Associated with Different Regions of the Human Colon Wall

The microorganisms associated with the undiseased human colon wall were examined in material obtained from four sudden-death victims. In traffic accident subjects (aged 45 and 16 years) the anaerobe-aerobe ratio was about 10⁴:1 in all areas of the colon examined, whereas in acute heart failure subjects (aged 74 and 46 years) the ratio was as low as 1.2:1. The flora of each individual was distinct and complex. Although the predominant anaerobes isolated were Bacteroides and Fusobacterium spp., which composed over 50% of the flora in some samples, the species isolated (indicated by morphology and glucose fermentation products) varied between individuals. Other major types observed were gram-positive nonsporing rods, including Bifidobacterium spp., and anaerobic cocci (between 8 and 20% of isolates). Clostridia were only isolated in significant numbers from one individual. Scanning electron microscopy showed that most of the organisms were present below the top surface of the mucin layer overlying the mucosa. The use of several different preparative procedures for microscopy showed a complex microbial structure within the mucus, but major variations in the bacterial populations in different areas of the colon were not found. Spiral-shaped organisms up to 60 μm long in the form of double helices were found in two subjects by scanning electron microscopy but were not isolated during the parallel bacteriological investigation. The differences between this and previous studies are discussed in relation to experimental procedures and also in contrast to results with animals that showed a particularly specialized flora associated with the colon wall.     

Microbial Flora of Irradiated Dungeness Crabmeat and Pacific Oysters

The microorganisms in Dungeness crabmeat (Cancer magister) and Pacific oysters (Crassostrea gigas) were identified by the replica-plating and computer analysis method. The initial flora of the shellfish and the flora change during storage at 7 C were determined. The microbial flora shifts in both shellfish were also determined after irradiation at 0.1 and 0.4 Mrad and during subsequent storage at 7 C. The Achromobacter species predominated in the initial flora of crabmeat (77.0%). The predominant position of this group increased to 99.2% after 0.1 Mrad and 100% after 0.4 Mrad. A large percentage of Lactobacillus was detected in oysters (55.0%). The Lactobacillus species were the predominant survivors after 0.1 Mrad (92.4%) but the predominant survivors after 0.4 Mrad were Achromobacter species (99.3%).