Molecular epidemiology of noroviruses associated with sporadic gastroenteritis in Guangzhou, China, 2013-2015.

Norovirus diarrhea is a great threat to public health worldwide. To characterize the prevalence of circulating noroviruses associated with sporadic gastroenteritis cases in Guangzhou, 215 stool specimens were collected during two consecutive cold seasons in 2013-2015. Noroviruses were detected in 25 (11.63 %) samples, and GII.4 (6/9) and GII.17 (10/16) were identified as the most predominant variants of each of those seasons. The remaining strains belonged to the genotypes GII.P12/GII.3, GII.2, and GI.Pb/GI.6. The phylogenetic relationships of the GII.17 strains were analyzed based on their capsid protein sequences. This study suggests a significant shift of predominant variants associated with sporadic gastroenteritis in Guangzhou.

Comparative genome analysis of a norovirus GII.4 strain GZ2013-L10 isolated from South China.

In this study, the genome sequence of a norovirus GII.4 strain isolated from South China was comparatively analyzed. The RNA genome of the strain GZ2013-L10 was composed of 7513 nucleotides. Phylogenetic analyses based on three ORFs confirmed its genotype as GII.Pe/GII.4-2012. Compared with other 22 genomes of the same variant, nine distinct nucleotide substitutions were found in the new genome, which resulted in three amino acid changes. All 138 capsid protein VP1 sequences of GII.4-2012 variants were also collected, and multiple alignments revealed 35 variable codons. Evolutionary analyses of GII.4-2012 variants were performed against previous pandemic GII.4 variants, and 2 distinctive changes were identified on epitopes A and E (E368, T413), which resulted in an obvious variation of their solvent-accessible surface areas. Therefore, the genome of GZ2013-L10 was extensively characterized, and new emerging variations on viral epitopes were predicted to contribute to NoV persistence in humans.

Prevalence and contamination patterns of Listeria monocytogenes in Flammulina velutipes plants.

Four mushroom (Flammulina velutipes) production plants were sampled to investigate the prevalence and contamination source of Listeria monocytogenes. Among 295 samples, the prevalence of L. monocytogenes was 18.6%; the contamination appeared to originate from the mycelium-scraping machinery, contaminating both the product and upstream packaging equipment. Of 55 L. monocytogenes isolates, lineages I.1 (1/2a-3a) and II.2 (1/2b-3b-7) accounted for 65.5% and 34.5%, respectively. In addition, lineage I.1 formed significantly thicker biofilms than those within lineage II.2, as determined by crystal violet staining and scanning electron microscopy. Genotype analyses of L. monocytogenes isolates using enterobacteria repetitive intergenic consensus-polymerase chain reaction, and random amplified polymorphic DNA revealed that the surfaces of mycelium-scraping machinery may serve as the main source of L. monocytogenes contamination in three of the four plants. This study was the first report to explore the potential contamination sources of L. monocytogenes in the mushroom production chain, thereby providing baseline information for adopting prophylactic measures for critical control points during production in mushroom plants to avoid L. monocytogenes contamination.

[Phenotypic and molecular characteristics of Salmonella enterica serotype 1,4, [5] ,12:i:--a review].

Salmonella enterica serotype 1, 4, [5 ], 12: i:- ( Salmonella 1, 4, [5], 12: i:-), an emerging serotype antigenically related to Salmonella Typhimurium (1,4, [5], 12 : i:1,2) but lacking the second phase flagellar antigen, has been frequently detected in many countries over the last 10 years. Nowadays it seems to be one of the major serotypes responsible for human salmonellosis cases worldwide. In addition, multidrug resistance is quite common in Salmonella 1, 4, [5],12:i:-, the two major clones (labelled as Spanish and European clones) show multidrug resistance to four or more unrelated classes of antimicrobials mediated by plasmids or chromosome. Some resistance determinants including bla(TEM), bla(CTX-M(-1), aac(3)-IV, aadA2, cmlA1, sul1, sul2, dfrA12, strA-strB, tet (A) and tet (B) have been found in these multidrug resistance strains. The genomic characterization of 1,4, [5] ,12:i:- isolates suggests that this serovar is likely to gather several clones or strains that have independently emerged from S. Typhimurium, and have changed through multiple independent events involving different clonal groups. In later study, emphasis should be paid on development of rapid and precise detection methods and study of pathogenic and resistance mechanisms of Salmonella 1,4, [5] ,12:i:-.

[Virulence-associated gene detection and ERIC-PCR typing of Campylobacter jejuni strains isolated from foods in four Southern Chinese provinces].

OBJECTIVE: To know food contamination and genetic diversity of Campylobacter jejuni in four provinces of South China, and to provide data for C. jejuni-associated foodborne disease prevention and control. METHODS: According to the national standard and the most probable number (MPN) method, we detected the contamination of C. jejuni from 558 food samples including vegetables, meat product, cooked food, seafood, frozen food, dairy product and edible fungi during 2011 and 2012. The isolates were used to detect 12 virulence-associated genes with PCR methods and construct ERIC-PCR fingerprints. RESULTS: Fourteen positive samples were determined from 558 samples, and all positive samples come from meat product samples. The average value of MPN of positive samples was 8.77 MPN/g. Virulence-associated gene analysis reveals that more than 50% of the C. jejuni isolates had at least 9 virulence genes. Interestingly, virB11 gene was not found and the genes of pldA and wlaN were 14.30% in all isolates. Total of 15 C. jejuni isolates could be divided into 10 genotypes belonging to 3 clusters by ERIC-PCR fingerprints. CONCLUSION: Meat product was the main source of C. jejuni food contamination in four provinces of South China. More control measures must be taken to avoid C. jejuni contamination.

[Numerical study on the performance effect of the ratio of long axis to short axis of upright polypropylene infusion bag].

The study aims to investigate the effect of the ratio of long axis to short axis (RLS) of upright polypropylene infusion bag on discharging process and to search the best RLS. Aiming at five different RLS (1. 5 : 1, 2 : 1, 3 : 1, 4 : 1 and 5 : 1, respectively) with the volume of 100 mL, 250 mL and 500 mL, respectively, based on finite element method, analyzing the variation of stress distribution, emptying rate, drugging space and steadiness coefficient, etc. For the bags of the same volume, emptying rate increased with increasing of RLS, but the steadiness coefficient decreased with increasing of RLS. The specific increasing amplitude of emptying rate and decreasing range of steadiness coefficient were as follows: 20% and 49% for 100 mL infusion bag, 9% and 51% for 250 mL infusion bag, and 11% and 46% for 500 mL infusion bag, respectibvely, when RLS increased from 1. 5 : 1 to 5 : 1. Comparatively speaking, the increasing amplitude of the emptying rate is remarkably less than the decreasing range of the steadiness coefficient. By comprehensive consideration of both emptying rate and steadiness coefficient, lower RLS is recommended for upright polypropylene infusion bag.

Complete genome analysis of a novel norovirus GII.4 variant identified in China.

The complete genome sequence of a novel norovirus strain GZ2010-L87 identified in Guangzhou was analyzed phylogenetically in this study. The RNA genome of the GZ2010-L87 strain is composed of 7,559 nucleotides. The phylogenetic analysis based on open reading frame (ORF) 2 revealed that the strain belongs to the GII.4 genotype, forming the new cluster GII.4-2009 which was also identified in Asia and the USA since 2009. Furthermore, phylogenetic analyses of the full genome and the different open reading frame sequences of GZ2010-L87 and other representative strains suggested that the novel strain did not undergo recombination. Comparative analysis with the consensus sequence of 31 completely sequenced norovirus GII.4-2009 genomes showed 86 mismatched nucleotides (56 in ORF1, 16 in ORF2, and 14 in ORF3), resulting in 19 amino acid changes (9 in ORF1, 3 in ORF2, and 7 in ORF3). Furthermore, 12 variable sites were found on the capsid protein of norovirus GII.4-2009, and most were located at the P2 domain. Meanwhile, based on comparison with other GII.4 clusters, 14 sites were shown specific to the novel cluster. In summary, the genome of the new GII.4-2009 variant GZ2010-L87, which was first identified in China, was extensively characterized with a large panel of genetically diverse noroviruses. The genomic information obtained from the novel variant can be used not only as a full-length norovirus sequence standard in China but also as reference data for future evolution research.

Sequencing of the grxB gene of Cronobacter spp. and the development of a PCR assay for its identification.

Cronobacter spp. (formerly Enterobacter sakazakii), a foodborne pathogen linked to powdered infant formula, is a rare cause of invasive infection with a high mortality rate in neonates. In this study, the Cronobacter sakazakii ATCC 29544 and C. muytjensii ATCC 51329 glutaredoxin 2 (grxB) genes were cloned and sequenced. Based on the unique regions of the Cronobacter grxB genes, two primers were synthesized to develop and optimize a Cronobacter-specific polymerase chain reaction (PCR) method. The PCR assay amplified a 378-bp DNA product from all positive controls, which are composed of 45 strains of Cronobacter spp., but not from any of 45 non-Cronobacter bacterial strains. The detection limits of this method are 10(4) colony-forming units (CFU)/mL of Cronobacter spp. in infant formula directly and 10(0) CFU/mL after an 8-h enrichment step. In summary, we have developed a PCR assay based on the grxB sequence. Combined with enrichment culturing, this technique offers a rapid and sensitive method for the detection of Cronobacter spp.

Genetic analysis of noroviruses associated with sporadic gastroenteritis during winter in Guangzhou, China.

Noroviruses are regarded as the major causes of acute gastroenteritis worldwide, but their prevalence in sporadic diarrhea in South China remains unclear. This study was performed to characterize the genotypes of circulating norovirus strains associated with sporadic diarrhea cases in Guangzhou from November 2010 to January 2011. Among fecal specimens collected from 89 patients with acute diarrhea, nine samples (10.11%) were norovirus positive and 32 samples (35.96%) were rotavirus positive. The partial polymerase and the capsid regions of these norovirus samples were sequenced and phylogenetically analyzed. Three genotypes (GII.4, GII.6, and GII.b/GII.3) were identified, among which GII.4-2006b was the most predominant genotype (4/9, 44.4%), followed by GII.6 (3/9, 33.3%). A novel GII.4-2010 variant was first detected in China. Furthermore, the near full-length genome of the GZ2010-L26 strain, which belonged to GII.4-2006b, was sequenced and analyzed. Thus, the results of this study suggested that, second to rotavirus, noroviruses are the important pathogens responsible for sporadic acute gastroenteritis during winter in Guangzhou, and the GII.4-2006b variant remains the predominant genotype.

[Isolation and identification of Cronobacter (Enterobacter sakazakii) strains from food].

OBJECTIVE: This study aimed to detect and quantify Cronobacter in 300 powdered milk samples and 50 non-powdered milk samples. Totally, 24 Cronobacter (formerly Enterobacter sakazakii) strains isolated from powdered milk and other foods were identified and confirmed. METHODS: Cronobacter strains were detected quantitatively using most probable number (MPN) method and molecular detection method. We identified 24 Cronobacter strains using biochemical patterns, including indole production and dulcitol, malonate, melezitose, turanose, and myo-Inositol utilization. Of the 24 strains, their 16S rRNA genes were sequenced, and constructed phylogenetic tree by N-J (Neighbour-Joining) with the 16S rRNA gene sequences of 17 identified Cronobacter strains and 10 non-Cronobacter strains. RESULTS: Quantitative detection showed that Cronobacter strains were detected in 23 out of 350 samples yielding 6.6% detection rate. Twenty-four Cronobacter strains were isolated from 23 samples and the Cronobacter was more than 100 MPN/100g in 4 samples out of 23 samples. The 24 Cronobacter spp. isolates strains were identified and confirmed, including 19 Cronobacter sakazakii strains, 2 C. malonaticus strains, 2 C. dubliensis subsp. lactaridi strains, and 1 C. muytjensii strain. CONCLUSION: The combination of molecular detection method and most probable number (MPN) method could be suitable for the detection of Cronobacter in powdered milk, with low rate of contamination and high demand of quantitative detection. 24 isolated strains were confirmed and identified by biochemical patterns and molecular technology, and C. sakazakii could be the dominant species. The problem of Cronobacter in powdered milk should be a hidden danger to nurseling, and should catch the government and consumer's attention.

Nitrogen-metabolising microorganism analysis in rapid sand filters from drinking water treatment plant.

Sand filters (SFs) are common treatment processes for nitrogen pollutant removal in drinking water treatment plants (DWTPs). However, the mechanisms on the nitrogen-cycling role of SFs are still unclear. In this study, 16S rRNA gene amplicon sequencing was used to characterise the diversity and composition of the bacterial community in SFs from DWTPs. Additionally, metagenomics approach was used to determine the functional microorganisms involved in nitrogen cycle in SFs. Our results showed that Pseudomonadota, Acidobacteria, Nitrospirae and Chloroflexi dominated in SFs. Subsequently, 85 high-quality metagenome-assembled genomes (MAGs) were retrieved from metagenome datasets of selected SFs involving nitrification, assimilatory nitrogen reduction, denitrification and anaerobic ammonia oxidation (anammox) processes. Read mapping to reference genomes of Nitrospira and the phylogenetic tree of the ammonia monooxygenase subunit A gene, amoA, suggested that Nitrospira is abundantly found in SFs. Furthermore, according to their genetic content, a nitrogen metabolic model in SFs was proposed using representative MAGs and pure culture isolate. Quantitative real-time polymerase chain reaction (qPCR) showed that ammonia-oxidising bacteria (AOB) and archaea (AOA), and complete ammonia oxidisers (comammox) were ubiquitous in the SFs, with the abundance of comammox being higher than that of AOA and AOB. Moreover, we identified a bacterial strain with a high NO3-N removal rate as Pseudomonas sp. DW-5, which could be applied in the bioremediation of micro-polluted drinking water sources. Our study provides insights into functional nitrogen-metabolising microbes in SFs of DWTPs.

Characterization of quinolone resistance in Salmonella enterica serovar Typhimurium and its monophasic variants from food and patients in China.

OBJECTIVES: The study aimed to characterize the quinolone resistance of Salmonella enterica serovar Typhimurium and its monophasic variant (Salmonella enterica serovar 1,4,[5],12:i:-) isolated from food and patients in China. METHODS: All of the isolates were assessed for quinolone susceptibility via the broth microdilution method. Then, the isolates were checked for mutations within quinolone resistance-determining regions of gyrA, gyrB, parC, and parE and were examined for plasmid-mediated quinolone resistance genes. RESULTS: High rates of resistance to nalidixic acid in the S. Typhimurium (70.7%) and S. 1,4,[5],12:i:- (41.9%) isolates were observed, and a considerable proportion of isolates with reduced susceptibility to ciprofloxacin and levofloxacin were also detected. The high frequency of mutations in GyrA (60.8%) and a variety of genes (aac[6']-Ib-cr [23.2%], oqxAB [19.2%], qnrS [13.6%], and qnrA [3.2%]) conferring quinolone resistance in these Salmonella isolates were noteworthy. Lastly, the isolates carrying qnrS for transferability and transmission of the quinolone resistance were analysed by conjugation. Multiple locus variable-number tandem repeat analysis profiles indicated that some qnrS-positive isolates were clonally related, whilst the other isolates were genetically divergent. This suggested that both clonal spread of resistant strains and horizontal transmission of the plasmid-mediated resistance genes contributed to the dissemination of qnrS-positive Salmonella isolates. CONCLUSION: This study highlights the prevalence of quinolone-resistant S. Typhimurium and S. 1,4,[5],12:i:- in China, posing a threat to public health.

Genomic Analysis and Stability Evaluation of the Phenol-Degrading Bacterium Acinetobacter sp. DW-1 During Water Treatment.

Phenol is a toxic organic molecule that is widely detected in the natural environment, even in drinking water sources. Biological methods were considered to be a good tool for phenol removal, especially microbial immobilized technology. However, research on the "seed" bacteria along with microbial community analysis in oligotrophic environment such as drinking water system has not been addressed. In this study, Acinetobacter sp. DW-1 with high phenol degradation ability had been isolated from a drinking water biofilter was used as seeded bacteria to treat phenol micro-polluted drinking water source. Meanwhile, the whole genome of strain DW-1 was sequenced using nanopore technology. The genomic analysis suggests that Acinetobacter sp. DW-1 could utilize phenol via the ß-ketoadipate pathway, including the catechol and protocatechuate branches. Subsequently, a bio-enhanced polyhedral hollow polypropylene sphere (BEPHPS) filter was constructed to investigate the stability of the seeded bacteria during the water treatment process. The denatured gradient gel electrophoresis (DGGE) profile and the quantification of phenol hydroxylase gene results indicate that when the BEPHPS filter was operated for 56 days, Acinetobacter sp. was still a persistent and competitive bacterium in the treatment group. In addition, 16S rRNA gene amplicon sequencing results indicate that Acinetobacter sp., as well as Pseudomonas sp., Nitrospira sp., Rubrivivax sp. were the predominant bacteria in the treatment group, which were different from that in the CK group. This study provides a better understanding of the mechanisms of phenol degradation by Acinetobacter sp. DW-1 at the gene level, and provides new insights into the stability of seeded bacteria and its effects on microbial ecology during drinking water treatment.

Highly efficient removal of Sb(V) from water by franklinite-containing nano-FeZn composites.

The existence of toxic and carcinogenic pentavalent antimony in water is a great safety problem. In order to remove antimony(V) from water, the purpose of this study was to prepare a novel graphene nano iron zinc (rGO/NZV-FeZn) photocatalyst via hydrothermal method followed by ultrasonication. Herein, weakly magnetic nano-Fe-Zn materials (NZV-FeZn, GACSP/NZV-FeZn, and rGO/NZV-FeZn) capable of rapid and efficient Sb(V) adsorption from water were prepared and characterised. In particular, rGO/NZV-FeZn was shown to comprise franklinite, Fe0, and graphite. Adsorption data were fitted by a quasi-second-order kinetic equation and Langmuir model, revealing that among these materials, NZV-FeZn exhibited the best Sb removal performance (543.9 mgSb gNZV-FeZn-1, R2 = 0.951). In a practical decontamination test, Sb removal efficiency of 99.38% was obtained for a reaction column filled with 3.5 g of rGO/NZV-FeZn. Column regenerability was tested at an initial concentration of 0.8111 mgSb L-1, and the treated water obtained after five consecutive runs complied with the GB5749-2006 requirement for Sb. rGO/NZV-FeZn was suggested to remove Sb(V) through adsorption-photocatalytic reduction and flocculation sedimentation mechanisms and, in view of its high cost performance, stability, and upscalable synthesis, was concluded to hold great promise for source water and wastewater treatment.

Prevalence, Virulence, Antimicrobial Resistance, and Molecular Characterization of Pseudomonas aeruginosa Isolates From Drinking Water in China.

Pseudomonas aeruginosa is an important opportunistic pathogen and remains a major threat to the microbial safety of drinking water. There is a lack of comprehensive data on P. aeruginosa contamination in drinking water in China. Therefore, this study aimed to determine the prevalence, genetic diversity, virulence genes, and antimicrobial resistance of P. aeruginosa isolated from mineral water and spring water in China. From January 2013 to January 2014, 314 drinking water samples were collected from 23 cities in China. Of the collected samples, 77 (24.5%) were contaminated with P. aeruginosa, and these comprised 34 raw water (30.4%), 39 activated carbon-filtered water (30.6%), and four final water product (3.9%). A total of 132 P. aeruginosa isolates were obtained, and all of them showed the presence of virulence genes, with the detection rates of ExoU, ExoS, phzM, toxA, and lasB genes being 7.6, 86.3, 95.5, 89.4, and 100%, respectively. All isolates were sensitive to the 14 antibiotics (ciprofloxacin, levofloxacin, ofloxacin, norfloxacin, gentamicin, tobramycin, amikacin, polymyxin B, imipenem, meropenem, aztreonam, ceftazidime, cefepime, and piperacillin/tazobactam) tested. The 132 isolates were categorized into 42 sequence types according to multilocus sequence typing, and ST235 accounted for 8.3% (11) of the total isolates. Thus, this study provides comprehensive data on the prevalence and characteristics of P. aeruginosa in drinking water in China and can aid in developing preventive measures against contamination during the drinking water treatment process.

Occurrence and Characterization of Fungi and Mycotoxins in Contaminated Medicinal Herbs.

Traditional medicinal herbs are widely used and may be contaminated with mycotoxigenic fungi during cultivation, harvesting, and storage, causing spoilage and mycotoxin production. We evaluated the predominant mycoflora and extent of mycotoxin contaminations in 48 contaminated samples of 13 different medicinal herbs. In total, 70.8% of herbs were slightly contaminated with aflatoxins (<5 µg kg-1). Codonopsis radix samples contained ochratoxin A (OTA) (360-515 µg kg-1), and Scutellariae radix samples contained OTA (49-231 µg kg-1) and citrinin (15-53 µg kg-1). Forty samples (83.3%) contained fungal contamination. Sixty-nine strains were characterized via morphological and molecular identification. The predominant mycoflora comprised four genera, Aspergillus spp. (26.1%), Penicillium spp. (24.6%), Rhizopus spp. (14.5%), and Trichoderma spp. (11.6%). Aflatoxins, OTA, and citrinin were detected in 37 cultures by high-performance liquid chromatography-tandem mass spectrometry. Approximately 21.6% of Aspergillus and Penicillium isolates produced mycotoxins. One Penicillium polonicum strain isolated from Scutellariae radix synthesized citrinin. Multiplex PCR analysis showed that three Aspergillus flavus strains harbored aflatoxin biosynthesis genes. One Aspergillus flavus strain isolated from Amomi fructus produced AFB1 and AFB2. To the best of our knowledge, the citrinin production by Aspergillus chevalieri and Penicillium sacculum was first reported in this study, which poses a potential risk of mycotoxin contamination in medicinal herbs.

Chromium ion removal from raw water by magnetic iron composites and Shewanella oneidensis MR-1.

In this study, nanoiron active carbon composites (NZVI/GAC) were used to remove chromium ions from raw water. The composites were synthesized from a novel formula of biological activated carbon and characterized by various techniques. The adsorption test data were fit by a pseudo-second-order kinetic model and Langmuir model. The qe and R2 values were 187 mg Cr/g and 0.9960, respectively, with 0.2 g/L NZVI/GAC at an initial concentration of 118 mg/L Cr according to the Langmuir isotherm model. Moreover, a Cr6+ detoxification reactor was constructed with the magnetic iron composite. The results indicated that the synthesized magnetic iron composite was a significant adsorbent for Cr6+ removal from aqueous solutions. The detoxification reactor was able to remove Cr6+ from raw water at an initial concentration of 26.5 mg/L within a short time period (3-5 min), with a removal efficiency of up to 99.90% and a treatment capacity of 45.0 mg Cr6+/g of adsorbent; the Cr6+ concentrations in the outflow met the GB5749-2006 requirements for drinking water. A synergistic effect between NZVI/GAC and a suspension of the bacterium Shewanella oneidensis MR-1 was found, showing that this bacterium can be used as a regeneration agent for iron-depleted activated carbon materials.

Composition and Dynamics of Bacterial Communities in a Full-Scale Mineral Water Treatment Plant.

The aim of this study was to gain insight into the bacterial composition and dynamics in a mineral water treatment system (MWTS). The bacterial community of a full-scale mineral water treatment plant in the Maofeng Mountain, South China, was studied using high-throughput sequencing combined with cultivation-based techniques in both the dry and wet season. Overall, adenosine tri-phosphate (ATP) concentration (6.47 × 10-11 - 3.32 × 10-8 M) and heterotrophic plate counts (HPC) (3 - 1.29 × 103 CFU/mL) of water samples in the wet season were lower than those (ATP concentration 5.10 × 10-11 - 6.96 × 10-8 M, HPC 2 - 1.97 × 103 CFU/mL) in the dry season throughout the whole MWTS. The microbial activity and biomass of water samples obviously changed along with treatment process. All 300 isolates obtained using cultivation-based techniques were distributed in 5 phyla, 7 classes, and 19 genera. Proteobacteria accounted for 55.7% (167) of the total isolates, among which predominant genus was Pseudomonas (19.3%). Illumina sequencing analysis of 16s rRNA genes revealed 15 bacterial phyla (relative abundance >0.1%) as being identified in all water samples. Among these, Proteobacteria constituted the dominant bacteria microbiota in all water samples. A large shift in the proportion of Bacteroidetes, Actinobacteria, and Firmicutes was obtained during the treatment process, with the proportion of Bacteroidetes, Actinobacteria decreasing sharply, whereas that of Firmicutes increased and predominated in the final water product. The core microbiome, which was still present in whole MWTS comprised several genera including Pseudomonas, Acinetobacter, Clostridium, and Mycobacterium, that contain species that are opportunistic pathogens, suggesting a potential threat for mineral water microbiology safety. This study is the first to investigate the bacterial community of a full-scale mineral water treatment plant in China. The results provided data regarding the bacteria composition and dynamics in an MWTS, which will contribute to the beneficial manipulation of the mineral water microbiome.

Isolation and Transcriptome Analysis of Phenol-Degrading Bacterium From Carbon-Sand Filters in a Full-Scale Drinking Water Treatment Plant.

Phenol is a typical organic contaminant in the environment. To date, the biodegradation of phenol by microorganisms remains the preferred method for its removal and remediation, but data on phenol removal by drinking water biofilters are lacking. In this study, we used high-throughput sequencing to investigate the microbial community structure in a carbon-sand biofilter. The results indicated that the predominant bacterial group was Bacilli, followed by Gammaproteobacteria, Clostridia, and Alphaproteobacteria. In addition, a strain was capable of degrading phenol at low concentrations of 500 µg/L within 100 min was isolated and identified as Rhodococcus sp. CS-1. Transcriptome analysis results showed that Rhodococcus sp. CS-1 was able to degrade phenol via both the catechol and protocatechuate branch of the ß-ketoadipate pathway. Furthermore, some novel candidate biomarkers (copper oxidase, copper chaperone, and MarR/DeoR/TetR family transcriptional regulators) were successfully identified to be potentially involved in phenol biodegradation. This study indicates that carbon-sand filters have the potential for remediation of phenol. The application of native microorganisms to drinking water treatment system is an adaptive strategy in oligotrophic water environments.

Prevalence and Molecular and Antimicrobial Characteristics of Cronobacter spp. Isolated From Raw Vegetables in China.

Cronobacter spp. is a foodborne pathogen that causes life-threatening and invasive diseases, such as necrotizing enterocolitis, meningitis, and sepsis. In this study, we aimed to investigate the prevalence, molecular characteristics and antimicrobial resistance of Cronobacter spp. in raw vegetables marketed in China. Based on dietary habits in China, 403 raw vegetables that could be eaten without additional cooking were collected. Of the 403 samples tested, 122 (30.27%) were positive for Cronobacter spp., and the contamination levels exceeded 110 most probable number (MPN)/g for 16.39% (20/122) of the samples. Coriander samples had the highest contamination rate of 52.81%, and the MPN values of 19.15% of positive coriander samples exceeded 100 MPN/g. Eleven serotypes were identified among 171 isolates, with Cronobacter sakazakii serogroup O1 (41 isolates) being the dominant serotype. Molecular characterization indicated that there was quite high genetic diversity in Cronobacter spp., and multilocus sequence typing analyses yielded 106 sequence types (STs), 55 of which were newly identified. Notably, the most prevalent ST (eight isolates) was C. malonaticus ST60, which appeared in a recent clinical infectious disease study in China. Five C. sakazakii ST4, seven C. malonaticus ST7, and three C. sakazakii ST8 confirmed as pathogenic STs in other countries were also detected in this study. Furthermore, all isolates were susceptible to amikacin, amoxicillin-clavulanic, cefepime, ciprofloxacin, and imipenem, but some isolates exhibited a high ratio of resistance to cephalothin (59.65%). In this study, the high contamination rate and the detection of pathogenic and new STs in raw vegetables indicated potential hazards to customers. To the best of our knowledge, this is the first report to provide valuable information on the contamination status of Cronobacter spp. in vegetables that can be eaten raw in China.