Number Of Bacterial Genera Research Articles

RETRACTED: Enhanced biofiltration coupled with ultrafiltration process in marine recirculating aquaculture system: Fast start-up of nitrification and long-term performance

In marine recirculating aquaculture systems (RAS), the biological nitrification system usually suffers from long time start-up and proliferation of potential pathogenic bacteria under the high salinity stress. This study aims to develop a strategy for fast start- up and long-term biosecurity of marine RAS by using a combined biofiltration system (ceramic rings (CR) - granular activated carbon (GAC) - ultrafiltration (UF) process) in an actual marine RAS for selenotoca multifasciata. The ammonia nitrogen and dissolved organic matter (DOM) removal performance, membrane fouling characteristics and microbial communities during long-term operation (200 days) were investigated. The CR-GAC-UF process minimized the start-up time of the nitrification system to 14 days under low ammonia concentrations (below 2 mg/L). Long-term operation of the marine RAS with ammonia and nitrite nitrogen stabilized below 0.1 mg/L with a daily ammonia nitrogen removal rate of 91.7 %. The UF membranes were operated at fluxes at 15 LHM and the protein/polysaccharide ratios of soluble microbial products (SMP) and extracellular polymeric substances (EPS) on the membranes were stable below 0.5 with low fouling levels. A number of bacterial genera that have the potential to degrade protein and humic-like substances were recognized, which was related to the long-term DOM removal. Candidatus_Nitrosopumilus, Nitrosomonas_sp. and Nitrospira sp. ENR4 were detected as the core nitrifiers in CR-GAC-UF process, which were closely related to the amoABC, hao and nxrA genes (p < 0.05). With the multi-stage barrier of CR-GAC-UF, the heterotrophic plate count in therecirculating water was kept at 3 ± 1 CFU/mL and the relative abundance of potential pathogens declined gradually below 0.81 %. Taken together, this study provides a theoretical basis for the wider application of marine RAS.

“Are we barking up the wrong tree? Too much emphasis on Cutibacterium acnes and ignoring other pathogens”— a study based on next-generation sequencing of normal and diseased discs

BACKGROUNDThe majority of literature on bacterial flora in the disc stands disadvantaged in utilizing traditional culture methods and targeting a single bacterium, Cutibacterium acnes. PURPOSEOur objective was to document the diversity in the bacterial flora between normal and degenerated discs for shortlisting potential pathogens using next-generation genomic tools. STUDY DESIGNExperimental case-control study. METHODSResearchers employed 16S metagenome sequencing to profile bacterial diversity in magnetic resonance imaging normal healthy discs from brain-dead organ voluntary donors (n=20) and 40 degenerated disc samples harvested during surgery (Modic [MC]=20 and non-Modic [NMC]=20). The V3-V4 region was amplified using universal bacterial primers 341F and 806R, and the libraries were sequenced using Illumina NovoSeq 6000 platform. Statistical significance was set at bacteria with a minimum of 100 operational taxonomic unit (OTU) and present in at least 70% of the samples. The quality check-filtered reads were processed using the QIIME-2 pipeline. The OTU clustering and taxonomic classification were carried out for the merged reads using the Greengenes/SILVA reference database. Validation was done by identification of bacterial metabolites in samples using the liquid chromatography–mass spectrometry approach. RESULTSAbundant bacteria differing widely in diversity, as evidenced by Alfa and Beta diversity analysis, were present in all control and degenerative samples. The number of bacterial genera was 27 (14-gram-positive: 13-gram-negative) in the control group, 23 (10-gram-positive: 11-gram-negative) in the Modic group, and 16 (11-gram-positive: 5-gram-negative) in the non-Modic group. In the Modic group, gram-negative bacteria OTUs were found to be predominant (more than 50% of the total bacteria identified), whereas in control and non-Modic groups the OTUs of gram-positive bacteria were predominant. Species-level analysis revealed an abundance of opportunistic gram-negative pathogens like Pseudomonas aeruginosa, Sphingomonos paucibacillus, and Ochrobactrum quorumnocens in the discs with Modic changes, more than in non-Modic discs. The presence of bacterial metabolites and quorum-sensing molecules like N-decanoyl-L-homoserine lactone, 6-hydroxynicotinic acid, 2-aminoacetophenone, 4-hydroxy-3-polyprenylbenzoate, PE (16:1(9Z)/18:0) and phthalic acid validated the colonization and cell-cell communication of bacteria in disc ruling out contamination theory. Cutibacterium acnes was not the predominant bacteria in any of the three groups of discs and in fact was in the 16th position in the order of abundance in the control discs (0.72%), seventh position in the Modic discs (1.41%), and 12th position (0.53%) in the non-Modic discs. CONCLUSIONThis study identified a predominance of gram-negative bacteria in degenerated discs and highlights that Cutibacterium acnes may not be the only degeneration-causing bacteria. This may be attributed to the environment, diet, and lifestyle habits of the sample population. Though the study does not reveal the exact pathogen, it may pave the way for future studies on the subject. CLINICAL SIGNIFICANCEThese findings invite further investigation into causal relationships of bacterial profile with disc degeneration phenotypes as well as phenotype-driven clinical treatment protocols.

Metagenomic Analysis of Gut Microbiome in Gout Patients with Different Chinese Traditional Medicine Treatments.

Introduction Changes in eating habits have made gout a metabolic disease of increasing concern. Previous studies have indicated that there are significant differences in species composition and abundance of gut microbiome in gout patients compared with average. Considering that traditional Chinese medicine has a momentous effect in treating gout, the research study aimed to explore the differences of genomic and metabolomics of gut microbiome before and after traditional Chinese medicine treatment in patients with gout. Method 30 patients with gout and 29 matched controls were recruited of which 16 patients took H treatment and 14 patients took T treatment. Stools were collected twice for patients before and after treatment and only once for controls. A total of 89 samples were annotated with metagenomic species and functions, and the enrichment analysis of differential genes and KO pathway was carried out. Result The results showed a decrease in the diversity of gut microbiome in gout patients and the gene abundance and metabolomics had great differences among study groups. The number of bacterial genera also had significant differences among treatment groups. Moreover, among different groups, the regulation of different species was variously correlated. The correlation between species and clinical laboratory indicators in the rising group was stronger than that in the decreasing group and the upregulation of some strain was related to the content of urea nitrogen. Conclusion After the traditional Chinese medicine treatment, the glutathione pathway was significantly enriched and some pathogenic bacteria were significantly inhibited. The study suggests that traditional Chinese medicine treatment may exert its therapeutic effect by inhibiting relevant pathways.

Effect of different fertilizers on the bacterial community diversity in rhizosperic soil of broomcorn millet (Panicum miliaceum L.)

ABSTRACT The bacterial community structure and diversity in the rhizosphere of broomcorn millet treated with different fertilizers were analyzed. The crops were grown under one of the four fertilization treatments: M1 (compound fertilizer), M2 (farmyard manure), M3 (biological bacterial fertilizer), and M4 (no fertilizer). The soil DNA was isolated, and the 16S rRNA library was sequenced using Illumina MiSeq sequencers. The bacterial diversity and the differential bacterial genera in four groups were analyzed. Compound fertilizers had the most significant effect on plant growth, followed by farmyard manure and then biological bacterial fertilizer (p < 0.05). Actinobacteria and Pseudomonas sp. were dominant in all soil samples. The abundance of Actinobacteria was higher in M1 and M4, and the abundance of Pseudomonas sp. was higher in M3 and M4. The number of bacterial genera (208) was the highest in M1. Eight bacterial genera with low abundance were common in M1 vs. M3 and M2 vs. M3. Four bacterial genera with high richness (Phormidium sp., Cellulosimicrobium sp., Yaniella sp., and Cloacibacterium sp.) overlapped in M1 vs. M3 and in M2 vs. M3. Compound fertilizer was the most effective at encouraging broomcorn millet growth (plant height, stem diameter, panicle weight, grain weight, and yield).

Effect of Polyvinyl Chloride Microplastics on Bacterial Community and Nutrient Status in Two Agricultural Soils.

Knowledge of the influence of microplastics on soil microbiome and nutrients is important for understanding the ecological consequences of microplastic pollution in terrestrial ecosystems. In this study, we investigated whether polyvinyl chloride (PVC) microplastic pollution at environmentally relevant concentrations would affect soil bacterial community and available nitrogen/phosphorus content. The results showed that although PVC microplastics at 0.1% and 1% levels did not have a significant effect on overall bacterial community diversity and composition in soil over the course of 35 days, a number of bacterial genera were significantly reduced or enriched by the presence of microplastics. Potentially due to their effect on certain functional groups, microplastics caused a significant change in soil available P content. It is noteworthy that, depending on soil type, pollution level and plasticizer presence, contrasting effects of microplastics may be observed. Further research is definitely warranted to gain a clearer picture of the threats posed by microplastic pollution in soil environments.

Root-associated microbiomes of wheat under the combined effect of plant development and nitrogen fertilization

BackgroundPlant roots assemble microbial communities both inside the roots and in the rhizosphere, and these root-associated microbiomes play pivotal roles in plant nutrition and productivity. Although it is known that increased synthetic fertilizer input in Chinese farmlands over the past 50 years has resulted in not only increased yields but also environmental problems, we lack a comprehensive understanding of how crops under elevated nutrient input shape root-associated microbial communities, especially through adjusting the quantities and compositions of root metabolites and exudates.MethodsThe compositions of bacterial and fungal communities from the roots and rhizosphere of wheat (Triticum aestivum L.) under four levels of long-term inorganic nitrogen (N) fertilization were characterized at the tillering, jointing and ripening stages. The root-released organic carbon (ROC), organic acids in the root exudates and soil organic carbon (SOC) and soil active carbon (SAC) in the rhizosphere were quantified.ResultsROC levels varied dramatically across wheat growth stages and correlated more with the bacterial community than with the fungal community. Rhizosphere SOC and SAC levels were elevated by long-term N fertilization but varied only slightly across growth stages. Variation in the microbial community structure across plant growth stages showed a decreasing trend with N fertilization level in the rhizosphere. In addition, more bacterial and fungal genera were significantly correlated in the jointing and ripening stages than in the tillering stage in the root samples. A number of bacterial genera that shifted in response to N fertilization, including Arthrobacter, Bacillus and Devosia, correlated significantly with acetic acid, oxalic acid, succinic acid and tartaric acid levels.ConclusionsOur results indicate that both plant growth status and N input drive changes in the microbial community structure in the root zone of wheat. Plant growth stage demostrated a stronger influence on bacterial than on fungal community composition. A number of bacterial genera that have been described as plant growth-promoting rhizobacteria (PGPR) responded positively to N fertilization, and their abundance correlated significantly with the organic acid level, suggesting that the secretion of organic acids may be a strategy developed by plants to recruit beneficial microbes in the root zone to cope with high N input. These results provide novel insight into the associations among increased N input, altered carbon availability, and shifts in microbial communities in the plant roots and rhizosphere of intensive agricultural ecosystems.

Zinc AA supplementation alters yearling ram rumen bacterial communities but zinc sulfate supplementation does not.

Despite the body of research into Zn for human and animal health and productivity, very little work has been done to discern whether this benefit is exerted solely on the host organism, or whether there is some effect of dietary Zn upon the gastrointestinal microbiota, particularly in ruminants. We hypothesized that (i) supplementation with Zn would alter the rumen bacterial community in yearling rams, but that (ii) supplementation with either inorganically sourced ZnSO4, or a chelated Zn AA complex, which was more bioavailable, would affect the rumen bacterial community differently. Sixteen purebred Targhee yearling rams were utilized in an 84-d completely randomized design, and allocated to one of three pelleted dietary treatments: control diet without fortified Zn (~1 × NRC), a diet fortified with a Zn AA complex (~2 × NRC), and a diet fortified with ZnSO4 (~2 × NRC). Rumen bacterial community was assessed using Illumina MiSeq of the V4 to V6 region of the 16S rRNA gene. One hundred and eleven OTUs were found with > 1% abundance across all samples. The genera Prevotella, Solobacterium, Ruminococcus, Butyrivibrio, Olsenella, Atopobium, and the candidate genus Saccharimonas were abundant in all samples. Total rumen bacterial evenness and diversity in rams were reduced by supplementation with a Zn AA complex, but not in rams supplemented with an equal concentration of ZnSO4, likely due to differences in bioavailability between organic and inorganically sourced supplement formulations. A number of bacterial genera were altered by Zn supplementation, but only the phylum Tenericutes was significantly reduced by ZnSO4 supplementation, suggesting that either Zn supplementation formulation could be utilized without causing a high-level shift in the rumen bacterial community which could have negative consequences for digestion and animal health.

Oral microbiota in autoimmune polyendocrine syndrome type 1

ABSTRACT Background: Autoimmune polyendocrine syndrome type-1 (APS-1) is a rare, childhood onset disease caused by mutations in the Autoimmune Regulator gene. The phenotypic expression is highly variable and includes disease manifestations in the oral cavity, including mucocutaneous candidiasis. Increasing evidence suggests a potential role of the skin, oral and gut microbiotas in the pathogenesis of autoimmunity. To date, no information exists regarding the oral microbiota in APS-1. Objective: To assess the bacterial microbiota of whole saliva in APS-1 patients by using high throughput sequencing. Design: Whole unstimulated saliva was collected from 10 APS-1 patients and 17 healthy controls and examined by high throughput sequencing of the hypervariable region V1-V2 of 16S rRNA using the 454 GS Junior system. Metastats (http://cbcb.umd.edu/software/metastats) was used to analyse the pyrosequencing reads. Results: A reduction in the total number of bacterial genera and species was detected in APS-1 compared to healthy controls. The proportion of the major phyla Firmicutes was higher (60% vs 41%, p = 0.002) and Bacteroidetes lower (15% vs 28%, p = 0.007) in APS-1 compared to healthy controls. On the genus level, Streptococcus and Gemella were prevalent in APS-1. Conclusion: Our findings indicate a significantly altered oral microbiota in APS-1.

Microbiological and bioinformatics analysis of primary Sjögren's syndrome patients with normal salivation§

BackgroundReduced salivation is considered a major clinical feature of most but not all cases of primary Sjögren's syndrome (pSS). Reduced saliva flow may lead to changes in the salivary microbiota. These changes have mainly been studied with culture that typically recovers only 65% of the bacteria present.ObjectiveThis study was to use high throughput sequencing, covering both cultivated and not-yet-cultivated bacteria, to assess the bacterial microbiota of whole saliva in pSS patients with normal salivation.MethodsBacteria of whole unstimulated saliva from nine pSS patients with normal salivation flow and from nine healthy controls were examined by high throughput sequencing of the hypervariable region V1V2 of 16S rRNA using the 454 GS Junior system. Raw sequence reads were subjected to a species-level, reference-based taxonomy assignment pipeline specially designed for studying the human oral microbial community. Each of the sequence reads was BLASTN-searched against a database consisting of reference sequences representing 1,156 oral and 12,013 non-oral species. Unassigned reads were then screened for high-quality non-chimeras and subjected to de novo species-level operational taxonomy unit (OTU) calling for potential novel species. Downstream analyses, including alpha and beta diversities, were analyzed using the Quantitative Insights into Microbial Ecology (QIIME) pipeline. To reveal significant differences between the microbiota of control saliva and Sjögren's saliva, a statistical method introduced in Metastats www.metastats.cbcb.umd.edu was used.ResultsSaliva of pSS patients with normal salivation had a significantly higher frequency of Firmicutes compared with controls (p=0.004). Two other major phyla, Synergistetes and Spirochaetes, were significantly depleted in pSS (p=0.001 for both). In addition, we saw a nearly 17% decrease in the number of genera in pSS (25 vs. 30). While Prevotella was almost equally abundant in both groups (25% in pSS and 22% in controls), about a twofold increase in pSS of Streptococcus (28% vs. 17%) and Veillonella (26% vs. 12%) was detected. Prevotella melaninogenica was the major species in controls (13%) while Veillonella atypica and the Veillonella parvula groups dominated in patient samples (14 and 14%). The scarcity in bacterial species in pSS compared with controls was also demonstrated by alpha and beta diversity analyses, as well as read abundance depicted in a phylogenetic tree.ConclusionsWhile Firmicutes was significantly higher in pSS patients than in controls, Synergistetes and Spirochaetes were significantly lower. The number of bacterial genera and species was also lower. These data showed that microbial dysbiosis is another key characteristic of pSS whole saliva which can occur independent of hyposalivation.

Down into the Earth: microbial diversity of the deepest cave of the world

AbstractIn our work, microbial diversity of Krubera-Voronja cave was evaluated in the view of the frequency of human visits in different locations as well as the sampling depth. Sampling in this cave was performed at depths of 220 m to 1640 m. Cultivation-independent method, namely barcoded pyrosequencing of 16S rRNA gene, was used for this analysis. Our results demonstrated high bacterial diversity at the phylum and genus levels. We have shown that the bacterial diversity at the phylum level depends on both the sampling depth and the frequency of human visits in Krubera-Voronja cave. Frequently visited locations were more diverse at the phylum level than the rarely visited branches. The total number of bacterial genera both per phylum and per sample correlated with the frequency of human visits but not with the sampling depth. Some genera, found in Krubera-Voronja cave, seem to be absent or very rare in other caves. The present study represents the first report on the microbial diversity in Krubera-Voronja cave

Association between Upper Digestive Tract Microbiota and Cancer-Predisposing States in the Esophagus and Stomach

The human upper digestive tract microbial community (microbiota) is not well characterized and few studies have explored how it relates to human health. We examined the relationship between upper digestive tract microbiota and two cancer-predisposing states, serum pepsinogen I/pepsinogen II ratio (PGI/II; predictor of gastric cancer risk) and esophageal squamous dysplasia (ESD; the precursor lesion of esophageal squamous cell carcinoma; ESCC) in a cross-sectional design. The Human Oral Microbe Identification Microarray was used to test for the presence of 272 bacterial species in 333 upper digestive tract samples from a Chinese cancer screening cohort. Serum PGI and PGII were determined by ELISA. ESD was determined by chromoendoscopy with biopsy. Lower microbial richness (number of bacterial genera per sample) was significantly associated with lower PGI/II ratio (P = 0.034) and the presence of ESD (P = 0.018). We conducted principal component (PC) analysis on a β-diversity matrix (pairwise difference in microbiota), and observed significant correlations between PC1, PC3, and PGI/II (P = 0.004 and 0.009, respectively), and between PC1 and ESD (P = 0.003). Lower microbial richness in upper digestive tract was independently associated with both cancer-predisposing states in the esophagus and stomach (presence of ESD and lower PGI/II). These novel findings suggest that the upper digestive tract microbiota may play a role in the etiology of chronic atrophic gastritis and ESD, and therefore in the development of gastric and esophageal cancers.

The underground organ microbial complexes of moorland spotted orchid &amp;lt;i&amp;gt;Dactylorhiza maculata&amp;lt;/i&amp;gt; (L.) So&amp;#243; (Orchidaceae)

This paper is focused on the first study of microbiota of a moderate climate orchid from Northern hemisphere—Moorland Spotted Orchid Dactylorhiza maculata (L.) Soó growing in its natural habitat. There have been obtained detailed data concerning bacterial communities from rhizosphere and inner tissues of young and old tubers. It was done using the biomarker analysis method where the markers were detected by gas chromatography—mass-spectrometry. It is shown that the number of bacterial genera and the bacteria amount (105 - 108 CFU per gram of dry weight) in D. maculata microbial complexes decreases from rhizosphere to old tuber to young. At the same time all three bacterial cenoses closely resemble each other in biodiversity. Their constant members are gg. Hyphomicrobium, Methylococcus, Nitrobacter, Pseudomonas and Sphingomonas (Proteobacteria), Bacillus and Clostridium (Firmicutes), Rhodococcus and Streptomyces (Actinobacteria). There were found differences peculiar to the taxonomic structure of each microbial complex. The rhizosphere is dominated by Actinobacteria while in inner tissues Proteobacteria are prevalent. The bacteria of gg. Ochrobactrum, Xanthomonas, Butyrivibrio, Corynebacterium, Mycobacterium, Propionibacterium, Sphingobacterium and specific iron reducers were shown to belong to rhizosphere community. Genera of Agrobacterium and Bifidobacterium were found only in the endophytic microbiota of the young tuber while g. Aeromonas is

Discovery and Analysis of Cofactor-dependent Phosphoglycerate Mutase Homologs as Novel Phosphoserine Phosphatases in Hydrogenobacter thermophilus

Phosphoserine phosphatase (PSP) catalyzes the dephosphorylation of phosphoserine to serine and inorganic phosphate. PSPs, which have been found in all three domains of life, belong to the haloacid dehalogenase-like hydrolase superfamily. However, certain organisms, particularly bacteria, lack a classical PSP gene, although they appear to possess a functional phosphoserine synthetic pathway. The apparent lack of a PSP ortholog in Hydrogenobacter thermophilus, an obligately chemolithoautotrophic and thermophilic bacterium, represented a missing link in serine anabolism because our previous study suggested that serine should be synthesized from phosphoserine. Here, we detected PSP activity in cell-free extracts of H. thermophilus and purified two proteins with PSP activity. Surprisingly, these proteins belonged to the histidine phosphatase superfamily and had been annotated as cofactor-dependent phosphoglycerate mutase (dPGM). However, because they possessed neither mutase activity nor the residues important for the activity, we defined these proteins as novel-type PSPs. Considering the strict substrate specificity toward l-phosphoserine, kinetic parameters, and PSP activity levels in cell-free extracts, these proteins were strongly suggested to function as PSPs in vivo. We also detected PSP activity from "dPGM-like" proteins of Thermus thermophilus and Arabidopsis thaliana, suggesting that PSP activity catalyzed by dPGM-like proteins may be distributed among a broad range of organisms. In fact, a number of bacterial genera, including Firmicutes and Cyanobacteria, were proposed to be strong candidates for possessing this novel type of PSP. These findings will help to identify the missing link in serine anabolism.

The Complex Vaginal Flora of West African Women with Bacterial Vaginosis

BackgroundThe spectrum of bacteria associated with bacterial vaginosis (BV) has recently expanded through taxonomic changes and the use of molecular methods. These methods have yet to be used in large-scale epidemiological studies in Africa where BV is highly prevalent.MethodsAn analysis of samples obtained during a clinical trial of the management of vaginal discharge in four West African countries. Samples were available from 1555 participants; 843 (54%) had BV. Nucleic acids of 13 bacterial genera or species potentially associated with BV were detected through the polymerase chain reaction.ResultsThe associations between various components of the vaginal flora were complex. Excluding Lactobacillus, the other 12 micro-organisms were all associated with each other at the p≤0.001 level. The prevalence of various bacterial genera or species varied according to age, sexual activity and HIV status. In multivariate analysis, the presence of Gardnerella vaginalis, Bifidobacterium, Megasphaera elsdenii, Dialister, Mycoplasma hominis, Leptotrichia, and Prevotella were independently associated with BV as was the absence of Lactobacillus and Peptoniphilus. However, Mobiluncus, Atopobium vaginae, Anaerococcus, and Eggerthella were not independently associated with BV. Unexpectedly, after treatment with a regimen that included either metronidazole or tinidazole, the proportion of patients with a complete resolution of symptoms by day 14 increased with the number of bacterial genera or species present at enrolment.ConclusionsNumerous bacterial genera or species were strongly associated with each other in a pattern that suggested a symbiotic relationship. BV cases with a simpler flora were less likely to respond to treatment. Overall, the vaginal flora of West African women with BV was reminiscent of that of their counterparts in industrialized countries.

Production of quorum-sensing-related signal molecules by epiphytic bacteria inhabiting wheat heads

The production of quorum-sensing-related signal molecules (QSRMs) among culturable bacteria comprising the community on wheat heads was investigated. The taxonomic position of 186 bacterial isolates obtained from ten heads was inferred based on 16S rRNA gene sequences, and their QSRM production was determined using two bioreporter strains of N-acylhomoserine lactones. Approximately 33% of isolates produced QSRMs, though the proportion of QSRM-producing isolates on a wheat head was significantly negatively correlated with population size. Most of the producing isolates were Pantoea species, most commonly Pantoea ananatis. Furthermore, the proportion of Pantoea ananatis that produced QSRMs was significantly negatively correlated with the number of bacterial genera (community richness) on each head. Finally, community richness was positively correlated with population size. Qualitative analysis using thin-layer-chromatography revealed that the QSRMs of Pantoea isolates were composed of at least two compounds. This is the first report indicating that Pantoea ananatis isolates inhabiting wheat heads are capable of producing QSRMs. QSRM production by Pantoea spp. may contribute to the predominance of this genus on wheat heads, particularly at relatively low population densities and community diversity.

Proinflammatory Cytokine and Nitric Oxide Induction in Murine Macrophages by Cell Wall and Cytoplasmic Extracts of Lactic Acid Bacteria

Cells from a number of bacterial genera have been shown to possess mitogenic and polyclonal activating properties when cultured with cells of the immune system. Based on previously reported health immune-enhancing effects of fermented dairy products, we tested the potentiating effects of representative lactic acid bacteria and their extracts on leukocyte function. Specifically, the effects of in vitro exposure to heat-killed cells of Bifidobacterium, Lactobacillus acidophilus, L. bulgaricus, L. casei, L. gasseri, L. helveticus, L. reuteri, and Streptococcus thermophilus, their cell walls, and their cytoplasmic extracts on proliferation as well as cytokine and nitric oxide (NO) production were examined in the RAW 264.7 macrophage cell line. A similar strategy was applied to murine cultures composed of peritoneal, spleen, and Peyer's patch cells. Both the cell wall and cytoplasmic fractions of lactic acid bacteria were able to stimulate cloned macrophages to produce significant amounts of tumor necrosis factor-α, (interleukin) IL-6, and NO. Pronounced enhancement of IL-6 production by peritoneal cells was observed when cultured with those extracts, whereas, effects were not noted in spleen and Peyer's patch cell cultures from mice. Based on the results, it appears that, as a group, the lactic acid bacteria were capable of stimulating macrophages and possibly other immune cells to produce cytokines and NO, and both their cell walls and cytoplasm contributed to these capacities.

Unusual Food-Borne Pathogens: Listeria Monocytogenes, Aeromonas, Plesiomonas, and Edwardsiella Species

Although these four groups of organisms are perceived as infrequent food-borne pathogens or of dubious significance, increasing epidemiologic data indicate that L. monocytogenes is an emerging cause of infections, particularly gastroenteritis. Furthermore, if data are ever generated that prove that most fecal isolates of Aeromonas are involved in bacterial diarrhea, then aeromonads will become recognized as important food-borne pathogens. For Plesiomonas and Edwardsiella, recognition of possible involvement in food-borne disease requires detailed medical histories, including foreign travel, contact with pets or animals, and food consumption histories.

The dual biosynthetic capability of N-acetylornithine aminotransferase in arginine and lysine biosynthesis.

The genes encoding the seven enzymes needed to synthesize L-lysine from aspartate semialdehyde and pyruvate have been identified in a number of bacterial genera, with the single exception of the dapC gene encoding the PLP-dependent N-succinyl-L, L-diaminopimelate:alpha-ketoglutarate aminotransferase (DapATase). Purification of E. coli DapATase allowed the determination of both the amino-terminal 26 amino acids and a tryptic peptide fragment. Sequence analysis identified both of these sequences as being identical to corresponding sequences from the PLP-dependent E. coli argD-encoded N-acetylornithine aminotransferase (NAcOATase). This enzyme performs a similar reaction to that of DapATase, catalyzing the N-acetylornithine-dependent transamination of alpha-ketoglutarate. PCR cloning of the argD gene from genomic E. coli DNA, expression, and purification yielded homogeneous E. coli NAcOATase. This enzyme exhibits both NAcOATase and DapATase activity, with similar specificity constants for N-acetylornithine and N-succinyl-L,L-DAP, suggesting that it can function in both lysine and arginine biosynthesis. This finding may explain why numerous investigations have failed to identify genetically the bacterial dapC locus, and suggests that this enzyme may be an attractive target for antibacterial inhibitor design due to the essential roles of these two pathways in bacteria.

Solving the problem of how to eat something as big as yourself: Diverse bacterial strategies for degrading polysaccharides

Polysaccharide digestion by bacteria is an important activity in many ecosystems, and a number of bacterial genera can perform this function. Although many papers have been published about the properties of isolated polysaccharide-degrading enzymes, relatively little is known about how intact bacteria degrade polysaccharides. This review summarizes recent findings suggesting that there are at least three different strategies. the most familiar one is the excretion of extracellular polysaccharidases, which diffuse to and degrade nearby polysaccharides. An example of this type of strategy is provided by the plant pathogen,Erwinia spp. A second strategy is to have the enzyme exposed to the extracellular medium but attached to the surface of the cell. Examples of this strategy are provided by the pullulanase system ofKlebsiella oxytoca and the cellulosomes ofClostridium thermocellum. A strategy that could be seen as a combination of the extracellular enzyme strategy and the surface organelle strategy is provided byVibrio harveyi, which attaches to its substrate, chitin, via proteins that appear to be specialized for attachment and produces extracellular enzymes that attack the chitin. A third strategy is to import the polysaccharide, as appears to be done byBacteroides spp. In this instance, the polysaccharide is bound to an outer membrane receptor, then passes into the periplasm where the degradative enzymes are located. The ecological advantages and disadvantages of these systems are discussed, and areas where further research is needed are defined.

Enantioselective hydrolysis of racemic methyl jasmonate using microorganisms and isolated enzymes

A variety of microorganisms were used to hydrolyze racemic methyl jasmonate [I] with varying degrees of enantioselectivity. The fungi tested included species from the genera Aspergillus, Penicillium, and Talaromyces. All fungi tested showed a preference for the [1 S,2 S( Z)]-(+)-isomer. The yeasts Saccharomyces cerevisiae and Candida albicans showed no activity. A number of bacterial genera were also tested. No activity could be shown for members of the genera Bacillus, Pseudomonas, Escherichia, Nocardia, and Thermoactinomyces. Hydrolytic activity was found in the genera Streptomyces and Mycobacterium. S. henetus showed the same enantioselectivity as the fungi, while M. phlei hydrolyzed the [1R,2R(Z)]-(−)-isomer preferentially. A number of isolated enzymes were also screened for activity. Varying degrees of hydrolytic activity and enantioselectivity were found.