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Purpose: Fusidic acid (FA), a potent steroidal antibiotic, is used topically to treat skin and soft tissue infections (SSTIs) caused by Staphylococci. The aim of this study is to report the prevalence of fusidic acid resistance among Staphylococcus epidermidis clinical isolates from a tertiary hospital in Wenzhou, east China. Methods: The antibiotic susceptibility of S. epidermidis isolates was determined by disc diffusion method and agar dilution method. Then, FA-resistant S. epidermidis isolates were characterized by multi-locus sequence typing, SCCmec typing and pulsed-field gel electrophoresis. Results: In the present study, the 55 (7.7%) FA-resistant S. epidermidis among 711 S. epidermidis clinical isolates were isolated from different parts of 53 patients. Fifty-five FA-resistant S. epidermidis isolates with FA MIC values ranged from 4 to 32 μg/mL. Among them, 50 (90.9%) were identified as methicillin-resistant Staphylococcus epidermidis (MRSE), in which mecA were positive. Meanwhile, the positive rates of fusB and fusC genes among FA-resistant S. epidermidis isolates were 85.5% (47/55) and 7.3% (4/55), respectively. All 55 isolates mentioned above were susceptible to vancomycin. More than 50% of FA-resistant isolates were resistant to non-β-lactam antimicrobials including erythromycin (80.0%, 44/55), clindamycin (65.5%, 36/55), ciprofloxacin (63.6%, 35/55) and sulfamethoxazole (63.6%, 35/55). A total of 14 sequence types (STs) were identified among the 55 FA-resistant S. epidermidis isolates, of which, ST2 (24/55, 43.6%) was the most predominant type. And the eBURST analysis showed that CC2, CC5 and CC247 accounted for 43.6% (24/55), 27.3% (15/55) and 14.5% (5/55), respectively. Meanwhile, a total of four SCCmec types (I, III, IV, V) were identified among the 55 FA-resistant S. epidermidis. Furthermore, the pulsed field gel electrophoresis divided the 55 isolates into 20 types, namely A-T. Q-type strains were most prevalent, accounting for 30.9% (17/55). Conclusion: Taken together, the dissemination of S. epidermidis ST2 clone with FA resistance can cause trouble in controlling S. epidermidis infections. [ABSTRACT FROM AUTHOR]

• Step-wise screening covers more than 600 known genotypes of thalassemia. • The first study to report a novel 4.9 Kb deletion at beta-globin gene. • Utilization of the third-generation sequencing in genotyping for thalassemia. Thalassemia is a common inherited haemoglobin disorder worldwide, several methods have been utilized in the step-wise screening. Even though hundreds of mutations in globin genes have been reported, novel mutations are continuously emerging as the development of DNA sequencing. The case is a 27-year-old female with abnormal values of routine hematological indices, who was admitted for genetic screening of thalassemia. Genomic DNA was extracted and used for genetic assays cover 26 mutations in HBA and HBB genes: gap-PCR and agarose gel electrophoresis were performed to detect deletions, while PCR-reverse dot blot was used to detect point mutations. The next- and third- generation sequencing were used to identify the known and potential novel genotypes of thalassemia, and multiplex ligation-dependent probe amplification (MLPA) was used for genotype validation. Hematological results indicate microcytic hypochromic anemia, high HbA2 (7.2%) and high HbF (6.2%). None of the known genotypes of thalassemia were matched for this case, but a novel 4.9 Kb deletion at HBB gene (hg38, Chr11: 5226187–5231089) was discovered by the third-generation sequencing, the novel deletion was also validated by MLPA (8 probes, 11p15.4: 203314–207652). This study suggests the third-generation sequencing has promising potentiality to discover novel genotypes (especially deletions) of thalassemia. [ABSTRACT FROM AUTHOR]

Salmonella enterica serotype Kentucky (S. Kentucky) is an important Salmonella serotype with multiple sequence types (ST) with a worldwide incidence. We identified 8 STs from 180 strains of S. Kentucky, and ST314 emerged as the most commonly encountered ST. Drug susceptibility testing revealed that ST314 had multiple resistance properties, and 75.5% of the strains were resistant to three or more classes of antimicrobials. The rate of resistance to chloramphenicol, florfenicol, sulfafurazole and tetracycline were greater than 60%. The rates of ST314 resistance to quinolones were as follows: ciprofloxacin, 32.1%; nalidixic acid, 16%; and ofloxacin, 7.5%. Investigating the mechanism of quinolone resistance of ST314 revealed that mutations in the quinolone resistance‐determining regions were rare, and resistance mainly occurred due to the resistance genes carried by plasmids. Only 1.9% (2/106) of ST314 strains had mutations in the quinolone resistance‐determining regions (QRDR). The drug resistance genes of ST314 were primarily of plasmid‐mediated quinolone resistance (PMQR). The detection rate of Salmonella genomic island 1 (SGI1) in ST314 was 12.3%. XbaI‐pulsed‐field gel electrophoresis revealed that S. enterica Kentucky ST314 was capable of cross‐regional and cross‐host transmission in China. We found ST314 to be the dominant S. Kentucky ST in China, and it carried multidrug resistance. This is the first report about the emergence of quinolone‐resistant S. enterica Kentucky ST314 in China, which is different from previous reports, and the findings of the present study suggest that the mechanism of quinolone resistance in these strains are plasmid‐mediated. Notably, plasmids carrying resistance genes may promote the rapid spread of ciprofloxacin resistance. [ABSTRACT FROM AUTHOR]

purpose of this study was to survey the prevalence, antimicrobial resistance, as well as the spread of resistance and virulence-associated gene of S. dysgalactiae. Methods: A total of 60 S. dysgalactiae strains were obtained from 830 milk samples from Holstein cows with clinical mastitis. Antimicrobial resistance was examined by the disk diffusion method. Antimicrobial resistance and virulence genes were investigated by PCR, agarose gel electrophoresis and 16S rRNA gene sequencing. Results: All isolates were resistant to tetracycline and showed a high level of resistance to aminoglycoside antibiotics, where 81.67% of the strains were multi-resistant to these ten sorts of antibiotics. In addition, the most prevalent resistance gene in S. dysgalactiae was aphA-1 (98.33%), followed by blaTEM (96.67%), ermB (83.3%), aadA1/aadA2 (78.33%) and tetL (73.33%). Totally, seven virulence genes with 25 combination patterns were detected in these isolates, and each isolates harbored at least one virulence gene. 21.67% of the isolates carried three or more virulence genes, while one strain with seven virulence-related genes and belonged to cfb+lmb+eno+napr+bca+scpB+cyl. Conclusion: These findings indicate that S. dysgalactiae isolated from clinical bovine mastitis cases in Northwest China show a variety of molecular ecology and are highly resistant to antibiotics commonly used in dairy farms. This research will help investigators better understand the pathophysiology S. dysgalactiae in bovine mastitis and choose the appropriate antibiotics to treat mastitis. [ABSTRACT FROM AUTHOR]

Background: The clearhead icefish, Protosalanx hyalocranius, is an economically important fishery species in China. Since 1980s, P. hyalocranius was widely introduced into lakes and reservoirs of northern China for aquaculture. However, the lack of a rapid and cost-effective sex identification method based on sex specific genetic markers has hindered study on sex determination mechanisms and breeding applications. Results: Female-specific genomic regions were discovered by comparing whole genome re-sequencing data of both males and females. Two female-specific genomic regions larger than 50 bp were identified, and one (598 bp) contained a putative FOXI gene, which was paralogous to another FOXI gene with sex-associated SNPs. The two FOXI sequences displayed significant length difference with nine deletions of total length of 230 bp. This deletion-type structural variation could be easily and efficiently detected by traditional PCR and agarose gel electrophoresis with one 569 bp band for males and two bands (569 and 339 bp) for females, which were validated in 50 females and 40 males with known phenotypic sexes. Conclusions: The results provided structural genomic evidence for the ZZ/ZW sex determination system in P. hyalocranius discovered in our previous study with association analysis of SNPs. Moreover, the female-specific markers and rapid and cost-effective PCR-based genetic sex identification method should have applications in further studies of sex determination mechanism for this species. [ABSTRACT FROM AUTHOR]

After the discovery of complete ammonia-oxidizing (comammox) Nitrospira , detection and assessments of the contribution of comammox Nitrospira communities to nitrogen cycling are in great demand. PCR-based approach, a common method for the detection of comammox, depends strongly on accurate amplification of the amoA genes from the original DNA samples using appropriate primers. In this study, we reported an evaluation of the performance of two commonly used primer sets, Ntsp-amoA 162F/359R and comaA/B-244f/659r, for amplifying the comammox amoA genes from three representative wetland soils in China [Sangsang (SS), Sanjiang (SJ), and Xianghai (XH)]. Our results demonstrated the two primer sets could both successfully amplify the clades with high relative abundances (RA), and further revealed a broadly similar diversity and community composition of dominant comammox operational taxonomic units (OTUs) (RA ≥ 1%) in each of the three wetland soils. However, the clades with low RA, such as the clade A (1.26%) in SJ and the clade B (11.54%) in XH that were recovered by metagenomics analysis, failed to be amplified using comaA/B-244f/659r, but were successfully amplified and sequenced using Ntsp-amoA 162F/359R. It indicated that, compared to comaA/B-244f/659r, Ntsp-amoA 162F/359R was more sensitive to the clades with low RA. However, it is worth noting that Ntsp-amoA 162F/359R would overestimate the RA of some rare clades. For example, the RAs of clade B in XH were overestimated by 32-fold. Furthermore, high levels of non-target amplification were detected via gel electrophoresis using both primer sets, especially for comammox Clade B amoA genes, implying that we should treat qPCR results based on these primers with caution. Taken together, our study comprehensively compared the performance of the two primer sets on the sensitivity and specificity of amplifying comammox amoA genes in three wetland soils, pointing out the necessity of further development of new primers for the efficient and accurate detection of comammox in various environments. [ABSTRACT FROM AUTHOR]

Background: Enterobacter cloacae is an opportunistic pathogen which is responsible for serious nosocomial infections. A gene which plays an important role in resistance to carbapenems is the New Delhi metallo-β-lactamase 1 (NDM-1). Currently, the spread of NDM-1-producing E. cloacae strains is a serious public threat. Methods: A multidrug-resistant E. cloacae ssp. dissolvens strain CBG15936 was recovered in 2017 in Guangzhou, China. PCR, S1-pulsed-field gel electrophoresis, and Southern blotting were performed to locate the bla NDM–1 gene. Susceptibility testing and conjugation experiments were also performed. Illumina HiSeq and Nanopore sequencers were used to perform whole-genome sequencing. Results: Strain CBG15936 belongs to ST932 and is resistant to carbapenems. The bla NDM–1 gene was found on a ∼62-kb plasmid, which has a conjugation frequency of 1.68 × 10–3 events per donor cell. Genome sequencing and analysis revealed that the NDM-1-carrying IncN1 plasmid contained a new transposon Tn 6696 , which consists of an intact qnrS1 -carrying Tn 6292 element, an inverted 8.3-kb Tn 3000 remnant, IS kpn19 , Δ tnpA , and IS 26. Conclusion: A new transposon, Tn 6696 , has been detected on a bla NDM–1-carrying plasmid recovered from multidrug-resistant E. cloacae ssp. dissolvens CBG15936 from China. This finding provides a new perspective regarding the potential for bla NDM–1 to undergo horizontal transfer among drug-resistant bacteria. [ABSTRACT FROM AUTHOR]

Background: Milk vetch dwarf virus (MDV) is an important ssDNA virus which causes yellowing, stunting and leaf rolling symptoms on legumes. In China, the virus causes great economic losses and has recently been found to infect tobacco. The expansion of its host range and its ability to spread rapidly has given rise to the urgent need for a sensitive, specific and rapid diagnostic assay that can assist in effective disease control. Methods: Assays based on the polymerase chain reaction combined with lateral flow strip detection (PCR-LFS) and recombinase polymerase amplification combined with LFS (RPA-LFS) were developed targeting the coat protein (CP) gene of MDV. Results: The PCR and RPA assays could detect respectively 103 copies or 101 copies of MDV by agarose gel electrophoresis. The PCR-LFS and RPA-LFS assays developed could both detect as few as 101 copies per reaction at 37 °C. Both methods could detect MDV in crude leaf extracts. Conclusions: The RPA-LFS assay developed is a rapid, sensitive and specific method for detecting MDV, which is convenient and has great potential for use in the field. [ABSTRACT FROM AUTHOR]

Background: The data on the prevalence of resistance to mupirocin (MUP), fusidic acid (FA) and retapamulin (RET) in methicillin-resistant Staphylococcus aureus (MRSA) from China are still limited. This study aimed to examine these three antibiotics resistance in 1206 MRSA clinical isolates from Eastern China. Phenotypic MUP, FA and RET resistance was determined by minimum inhibitory concentrations (MICs), and genotypic by PCR and DNA sequencing of the mupA/B, fusB-D, cfr, vgaA/Av/ALC/B/C/E, lsaA-C/E and salA and mutations in ileS, fusA/E, rplC, and 23S RNA V domain. The genetic characteristics of resistance isolates were conducted by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results: Overall MRSA MUP, FA and RET resistance was low (5.1, 1.0 and 0.3%, respectively). MupA was the mechanism of high-level MUP resistance. All low-level MUP resistance isolates possessed an equivocal mutation N213D in IleS; of these, 2 reported an additional V588F mutation with an impact on the Rossman fold. FusA mutations, such as L461K, H457Q, H457Y and V90I were the primary FA mechanisms among high-level resistance isolates, most of which also contained fusC; however, all low-level resistance strains carried fusB. Except lsaE gene detected in one isolate, no other resistance mechanisms tested were found among RET-resistant isolates. Additionally, sixteen PFGE types (A-P) were observed, among which type B was the most common (49/76, 64.5%), followed by types E and G (4/76, 5.3% each) and types C and M (3/76, 3.9% each). All resistant strains were divided into 15 ST types by MLST. ST764 (24/76, 31.6%), ST630 (11/76, 14.5%), ST239 (9/76, 11.8%) and ST5 (7/76, 9.2%) were the major types. PFGE type B isolates with the aforementioned STs were mainly found in mupirocin resistant isolates. Conclusions: MUP, FA and RET exhibited highly activity against the MRSA isolates. Acquired genes and chromosome-borne genes mutations were responsible for MUP and FA resistance; however, the mechanism for some RET-resistant isolates remains to be further elucidated. Also, the surveillance to MUP in MRSA should be strengthened to prevent elevated resistance due to the expansion of clones. [ABSTRACT FROM AUTHOR]

This study evaluated the effects of starter cultures (Lactobacillus sakei and Staphylococcus xylosus) on lipid oxidation and accumulation of biogenic amines (BAs) in Chinese smoked horsemeat sausage. Physicochemical parameters, microorganisms, and sensory properties were assessed in smoked horsemeat sausage. Traditional microbiological detection and polymerase chain reaction‐denaturing gradient gel electrophoresis analysis showed that the starter cultures inhibited the growth of pathogenic bacteria in smoked horsemeat sausage during ripening. In addition, the starter cultures not only significantly reduced lipid oxidation, but also inhibited the accumulation of BAs (cadaverine, tyramine, putrescine, 2‐phenylethylamine, histamine, and tryptamine) in smoked horsemeat sausage. Sensory evaluation results showed that the starter cultures significantly improved the color, taste, odor, and overall acceptability scores of the smoked horsemeat sausage. Overall, the starter cultures were suitable for production of smoked horsemeat sausage with good organoleptic qualities. Practical applications: In China, smoked horsemeat sausage is one of the most popular traditional fermented sausage with a long history, but it is difficult to control quality during product fermentation. However, to the best of our knowledge, few studies exist on the effect of mixed starter cultures on smoked horsemeat sausage. This study strongly evidenced that the mixed starter cultures (Lactobacillus sakei and Staphylococcus xylosus) can significantly inhibit lipid oxidation and the accumulation of BAs in smoked horsemeat sausage, thereby improving product quality. [ABSTRACT FROM AUTHOR]

Copyright of China Brewing is the property of China Brewing Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Proteus mirabilis is an opportunistic pathogen that causes diarrhea and urinary tract infections in humans and companion animals. Emergence of antimicrobial resistance in P. mirabilis increases risk of failure of antimicrobial therapy in companion animals. The current study determined prevalence, genetic diversity and phenotypic antimicrobial resistance of P. mirabilis isolated from feces of 35 dogs with diarrhea hospitalized in Beijing, China. Random amplified polymorphic DNA (RAPD), pulse-field gel electrophoresis (PFGE), PCR and antimicrobial disk diffusion were used to characterize these isolates. Prevalence of P. mirabilis in feces from hospitalized dogs was 28%. RAPD and PFGE demonstrated a great diversity of isolates. Thirteen (37%) of 35 isolates produced extended-spectrum beta-lactamases (ESBLs), with blaTEM being detected in all ESBLs-producing isolates. All isolates were susceptible to imipenem, cefoxitin and cefotaxime/clavulanic. rmtB (with 51% prevalence rate) was the only aminoglycoside 16S rRNA methylase resistance gene identified. Among plasmid-mediated quinolones resistance genes, qnrB (14%) and aac(6')-Ib (26%) were detected. In conclusion, P. mirabilis isolates from feces of dogs with diarrhea in China had great genetic diversity and a high prevalence produced ESBLs. Consequently, antimicrobial stewardship programs should also target companion animals to reduce emergence of antimicrobial resistance. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Southern Agriculture is the property of Journal of Southern Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

BACKGROUND: Chinese local buffalos are mainly used as draft animals because of their low productivity but their crossbreeding with Murrah and Nili‐Ravi breeds can produce offspring with a greatly improved milk yield. However, no studies have reported the characteristics of milk produced during lactation by these crossbred buffalo. RESULTS: This study investigated changes in yield, and the physicochemical composition of milk of Murrah, Nili‐Ravi, Murrah×local, Murrah×Nili‐Ravi×local buffalos with milk yields of 1022.49 ± 90.26 kg, 1193.02 ± 97.65 kg, 805.46 ± 76.57 kg and 1499.35 ± 105.32 kg respectively over a 210‐day period. The protein, fat, ash and total solids contents of milk from the hybrids decreased, but the yield of these nutritional components increased because of the greater improvement in milk yield. As lactation progressed, lactose content increased, but this change was not significant after the 15th day postpartum; the protein content decreased whereas fat content increased significantly during the first three days; ash content changed significantly during the first 24 h postpartum but further changes were not significant. Gel electrophoresis was used to identify the protein profile with no difference found between the four breeds. CONCLUSION: After crossbreeding, the milk production of triple‐crossbred buffalo was higher than Murrah, Nili‐Ravi and the local Chinese buffalo, making it a potential resource for the Chinese dairy market. The results of this experiment will provide basic data for making better use of buffalo milk, planning crossbreeding programs, and establishing standards for buffalo milk. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

An increasing number of severe infections caused by Staphylococcus aureus ST398 strains has been observed. However, it has not been elucidated whether all ST398 strains are equally virulent. We collected 13 strains from China and Canada to test in a Caenorhabditis elegans infection model and compared their whole genome sequences (WGS) to explore potential insights into their virulence. All isolates belonged to ST398-methicillin-susceptible S. aureus (MSSA) with variant spa types (t034, t571, t1451, t1250). Pulsed field gel electrophoresis (PFGE) and WGS analyses showed that the 13 isolates clustered into 3 genomic types (Types A-C). WGS and prophage phylogenetic analyses also revealed that the strains could be divided into 3 phage groups (Groups 1–3), which correlated with high-, moderate-, and low-nematocidal activities, with mean killing rates of 94, 67, and 40%, respectively. Group 1 carried ϕSa3-Group 1 (ϕSa3-G1), Group 2 carried ϕSa3-G2, and Group 3 lacked ϕSa3. Interestingly, strain GD1706 (that genetically clustered within Type C) and strain GD487 (within Type B) both carried ϕSa3-G1 like phages and killed 92% of the nematodes, similar to the Type A strains carrying ϕSa3-G1. This study demonstrated that different ST398 sub-lineages possess variable virulence capacities, depending on the presence or absence, as well as the structure of the prophage ϕSa3 that carries virulence factors. IMPORTANCE: Since first being reported in the early 2000s, Staphylococcus aureus ST398 has not only become recognized as a frequent colonizing strain in economically important livestock animals, but has also proven to be a concern for infection in humans and, in particular, has been linked to higher rates of severe invasive human infections. We collected ST398 strains from China and Canada to test in a worm (Caenorhabditis elegans) infection model and compared their whole genome sequences to gain insight into pathogenesis. We have shown that different ST398 sub-strains differ in their virulence potential based on the presence or absence and structure of prophage ϕSa3, which carries important virulence factors. Our observations suggest that ST398 strains are relatively heterogeneous from a clinical perspective, and more studies are needed to differentiate between virulent and non-virulent ST398 strains to determine the true global spread of relevant sub-strains. [ABSTRACT FROM AUTHOR]

Bartonella are vector-borne hemotropic bacteria that infect a wide variety of hosts, including people. While there are PCR assays that can identify individual or groups of Bartonella, there is no reliable molecular method to simultaneously detect all species while maintaining genus specificity and sensitivity. By comparing highly conserved 16S rRNA sequences of the better-recognized Bartonella spp. on GenBank, we selected primers and probes for a genus-specific pan-Bartonella FRET-qPCR. Then, a gltA-based Bartonella PCR was established by selecting primers for a highly variable region of gltA, of which the sequenced amplicons could identify individual Bartonella spp. The pan-Bartonella FRET-qPCR did not detect negative controls (Brucella spp., Anaplasma spp., Rickettsia spp., Coxiella burnetii, and Wolbachia) but reliably detected as few as two copies of the positive control (Bartonella henselae) per reaction. There was complete agreement between the pan-Bartonella FRET-qPCR and the gltA-based Bartonella PCR in detecting Bartonella in convenience test samples from China and St. Kitts: cats (26%; 81/310), Ctenocephalides felis (20%; 12/60), cattle (24%; 23/98), and donkeys (4%; 1/20). Sequencing of the gltA-based Bartonella PCR products revealed B. henselae (70%; 57/81) and B. clarridgeiae (30%; 24/81) in cats and C. felis (67%; 8/12, and 33%; 4/12, respectively) and B. bovis in cattle (23.5%; 23/98) and donkeys (4.0%; 1/24). The pan-Bartonella FRET-qPCR and gltA-based Bartonella PCR we developed are highly sensitive and specific in detecting recognized Bartonella spp. in a single reaction. The pan-Bartonella FRET-qPCR is convenient requiring no gel electrophoresis and providing copy numbers, while the gltA-based Bartonella PCR reliably differentiates individual Bartonella species. The use of these PCRs should greatly facilitate large-scale surveillance studies and the diagnosis of infections in clinical samples. [ABSTRACT FROM AUTHOR]

Purpose: Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae bacteria causes nosocomial infections worldwide. However, KPC-producing K. pneumoniae outbreak has never been reported in Shandong Province, China. The purpose of our study was to elucidate the epidemiological and drug resistance mechanisms of KPC-producing K. pneumoniae strains collected from a large teaching hospital in Shandong during the outbreak. Moreover, we attempted to characterize the genetic environment and phylogenetic analysis of blaKPC-2 in outbreak isolates. Methods: We monitored a 64-day outbreak of infection in a general hospital in Shandong Province, and the bacteria causing the infection were all ST11-type K. pneumoniae. The genotype correlation of KPC-producing K. pneumoniae isolates was assessed by whole-genome sequencing (WGS) phylogenetic analysis. Subsequent studies included antibiotic susceptibility testing, multilocus sequence typing (MLST) and S1-pulsed-field gel electrophoresis (S1-PFGE), Southern blot hybridization. Results: From February 1, 2018 to April 5, 2018, 14 KPC-producing K. pneumoniae isolates from different wards were collected. All 14 isolates were resistant to carbapenems and carried the extended-spectrum β-lactamase (ESBL) gene as well as fosA, and sul genes. Whole-genome analysis showed that all 14 the outbreak isolates were all ST11 type. The blaKPC-2 carrying plasmids were all belong to IncFIIK2 type, and the size ranged from 94 kb to 368 kb. Conclusion: As far as we know, this report first describes the genomics characterization of KPC-2-producing K. pneumoniae outbreak isolates from Shandong Province, China. In our study, these isolates appeared to be cloned, and ST11 K. pneumoniae was the major clone caused the outbreak. Therefore, routine surveillance of such strains in this region is urgently warranted. [ABSTRACT FROM AUTHOR]

Thirty-five serotype K1 hypervirulent Klebsiella pneumoniae (K1-hvKP) isolates collected from a Chinese hospital during the whole year of 2017 were evaluated to characterize the prevalence of the plasmid-mediated quinolone resistance (PMQR) genes. In total, 18 (51.4%) isolates were detected to carry PMQR genes, and the most frequently detected gene was qnrS1 (37.5%), followed by aac(6′)-Ib-cr (15%) and qnrB4 (2.5%). Remarkably, some qnr-carrying strains had only a single or more quinolone resistance-determining region mutations in GyrA or ParC, and most exhibited high-level ciprofloxacin resistance. However, only 11.4% (4/35) isolates were resistant to quinolones. Furthermore, 34.3% (12/35) carried extended-spectrum β-lactamase (ESBL) genes, but only 14.3% (5/35) exhibited ESBL phenotype. The most prevalent virulence genes were mrkD (100%, 21/21), followed by magA (97.1%, 34/35), wabG (97.1%, 34/35), rmpA (97.1%, 34/35), aerobactin (94.3%, 33/35), kfuB (94.3%, 33/35), ycf (91.4%, 32/35), iutA (91.4%, 32/35), rmpA2 (62.9%, 22/35), wcaG (62.9%, 22/35), ybtS (51.4%, 18/35), allS (45.7%, 16/35), and iroN (22.9%, 8/35). Multilocus sequence typing (MLST) analysis assigned the 35 K1-hvKP isolates into 5 sequence types (STs), with ST23 encompassing 77.1% of the strains. Pulsed field gel electrophoresis (PFGE) typing showed that strains closely related by MLST clustered in major PFGE clusters, of which cluster A accounts for 16 ST23 isolates and cluster B includes 11 ST23 isolates. The analysis of the transconjugants showed decreased susceptibility to quinolones and revealed a cotransfer of blaCTX-M-15 with the qnrS1 alleles. Cumulatively, our findings showed that the PMQR-producing K1-hvKP strain is covertly spreading even when K1-hvKP is rarely resistant to fluoroquinolones (FQs) according to the Clinical and Laboratory Standards Institute criteria. It is therefore critical to continuously monitor the PMQR-producing K1-hvKP epidemiology and minimize potential risks from FQ-resistant K1-hvKP. [ABSTRACT FROM AUTHOR]

The increasing prevalence and transmission of the carbapenem resistance gene bla NDM–5 has led to a severe threat to public health. So far, bla NDM–5 has been widely detected in various species of Enterobacterales and different hosts across various cities. However, there is no report on the bla NDM–5– harboring Morganella morganii. In January 2016, the first NDM-5-producing Morganella morganii L241 was found in a stool sample of a patient diagnosed as recurrence of liver cancer in China. Identification of the species was performed using 16S rRNA gene sequencing. Carbapenemase genes were identified through both PCR and sequencing. To investigate the characteristics and complete genome sequence of the bla NDM–5-harboring clinical isolate, antimicrobial susceptibility testing, S1 nuclease pulsed field gel electrophoresis, Southern blotting, transconjugation experiment, complete genome sequencing, and comparative genomic analysis were performed. M. morganii L241 was found to be resistant to broad-spectrum cephalosporins and carbapenems. The complete genome of L241 is made up from both a 3,850,444 bp circular chromosome and a 46,161 bp self-transmissible IncX3 plasmid encoding bla NDM–5, which shared a conserved genetic context of bla NDM–5 (ΔIS 3000 -ΔIS Aba125 -IS 5 - bla NDM–5- ble - trpF - dsbC -IS 26). BLASTn analysis showed that IncX3 plasmids harboring bla NDM genes have been found in 15 species among Enterobacterales from 13 different countries around the world thus far. In addition, comparative genomic analysis showed that M. morganii L241 exhibits a close relationship to M. morganii subsp. morganii KT with 107 SNPs. Our research demonstrated that IncX3 is a key element in the worldwide dissemination of bla NDM-5 among various species. Further research will be necessary to control and prevent the spread of such plasmids. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is an important foodborne pathogen with a significant impact on public health worldwide. A great number of outbreaks caused by L. monocytogenes has been reported, especially in the United States, and European countries. However, listeriosis has not yet been included in notifiable disease in China, and thus information on this infection has been scarce among the Chinese population. In this study, we described a 3-year surveillance of listeriosis in Beijing, China. Fifty-six L. monocytogenes strains isolated from 49 clinical infectious cases (27 pregnancy-associated infections and 22 non-pregnancy-associated infections) were analyzed by serotyping, pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and antimicrobial susceptibility testing between 2014 and 2016 in Beijing. The predominant serogroups were 1/2a,3a and 1/2b,3b,7 which accounted for 92% of the overall isolates. Four strains were serogroup 4b,4d,4e, isolated from patients with pregnancy-associated infections. Based on PFGE, these isolates were divided into 32 pulsotypes (PTs) and 3 clusters associated with serogroups. Ten PTs were represented by more than one isolate with PT09 containing the most number of isolates. MLST differentiated the isolates into 18 STs, without new ST designated. The three most common STs were ST8 (18.4%), ST5 (16.3%), and ST87 (12.2%), accounting for 46.9% of the isolates. STs prevalent in other parts of the world were also present in China such as ST1, ST2, ST5, ST8, and ST9 which caused maternal fetal infections or outbreaks. However, the STs and serogroup distribution of clinical L. monocytogenes in Beijing, China was different from those in other countries. Strains of ST1 and ST2 were isolated from patients with pregnancy-associated infection, whereas none of ST155 isolates caused pregnancy-associated cases. Surveillance of molecular characterization will provide important information for prevention of listeriosis. This study also enhances our understanding of genetic diversity of clinical L. monocytogenes in China. [ABSTRACT FROM AUTHOR]

Background: As a result of the growing prevalence of the plasmid-mediated mobile colistin resistance gene mcr-1 among Gram-negative bacteria, the surveillance of mcr-1 has been globally applied. In our study, we aimed to shed light on the possibility of transmission of mcr-1-resistant isolates through market retail fruits. Methods and results: Herein, 133 different fruit surface samples were collected and screened for the different MCR variants (mcr-1 to mcr-8) using PCR and confirmed with sequencing. We identify for the first time mcr-1-carrying Escherichia coli and Klebsiella pneumoniae from market retail fruits in Guangzhou, China. Minimum inhibitory concentrations were detected by the broth microdilution method. Liquid mating was performed to check the transferability of the mcr-1 gene. Pulsed field gel electrophoresis analysis of S1 nuclease-digested DNA and Southern blotting were performed to check the location of the mcr-1 gene. Then, whole-genome sequencing and in silico multilocus sequence typing analysis were performed. Conclusion: We showed that E. coli GB110 can mediate the spreading of antibiotic resistance genes through the food chain, while K. pneumoniae GB015 was considered to be the progenitor of the most successful multidrug-resistant clone. Since fruits are usually consumed fresh, this may serve as a direct source of mcr-1-producing bacteria in humans that requires prompt surveillance and intervention to limit the spread of resistance. [ABSTRACT FROM AUTHOR]

A low phytic acid (lpa) rice mutant line, denoted as Os-lpa-XS110-2 , was developed through gamma irradiation (Liu et al., 2007). Subsequently, this mutant line was cultivated alongside its parental wild-type counterpart, Xiushui 110, in four distinct field trials conducted in China. The primary objective of this research was to conduct a comparative proteomic analysis between the parental and mutant lines, aiming to detect and compare protein species with varying levels of relative abundance and their potential involvement in phytic acid biosynthesis. To elucidate the molecular underpinnings of this alteration, proteome analyses were conducted on both the parental and mutant lines, employing LC-MS/MS in conjunction with 2-dimensional gel electrophoresis. Only, 7 proteins consistently exhibited significant differential abundance across all trial locations, suggesting that the regulation of these proteins is less susceptible to environmental influences. In a broader context, among the 700 detected proteins, 56 displayed noteworthy up- or down-regulation across diverse geographical regions. Notably, storage and housekeeping proteins emerged as the most frequently identified protein categories, with globulin and glutelin demonstrating prominent differences in abundance. However, based on our current understanding, none of the identified proteins appear to have direct roles in phytic acid biosynthesis. [Display omitted] • Research identified differentially expressed proteins in low-phytic mutant rice. • Among these proteins, globulin and glutelin were predominantly abundant. • This may be of importance to both the mutation event and growth conditions. • Consistent differences between natural biological variation and mutation were sought. • Screening proteome alterations may contribute to the safety assessment of mutants. [ABSTRACT FROM AUTHOR]

Background: Little is known about the association of the single nucleotide polymorphism (SNP) of rs364585 near serine palmitoyl-transferase long-chain base subunit 3 gene (SPTLC3) and serum lipid profiles. The present study was detected the association of the SPTLC3 rs364585 SNP and several environmental factors with serum lipid profiles in the Han and Jing populations. Methods: Genotyping of the SPTLC3 rs364585 SNP was performed in 824 unrelated individuals of Han and 783 participants of Jing by polymerase chain reaction and restriction fragment length polymorphism combined with gel electrophoresis, and then confirmed by direct sequencing. Results: There was no significant difference in either genotypic or allelic frequencies between Han and Jing, or between males and females of the both ethnic groups. The levels of serum low-density lipoprotein cholesterol (LDL-C) and the ratio of apolipoprotein (Apo) A1 to ApoB in Han; and total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and LDL-C in Jing were different between the A allele carriers and the A allele non-carriers (P < 0.05-0.001). Subgroup analysis according to sex showed that the levels of LDL-C in Han males; TC and LDL-C in Jing males; and HDL-C and LDL-C in Jing females were different between the A allele carriers and the A allele non-carriers (P < 0.05-0.001), the A allele carriers had higher LDL-C and TC levels, and lower HDL-C levels than the A allele non-carriers. Serum lipid traits were also associated with several environmental factors in the Han and Jing populations, or in males and females of the both ethnic groups. Conclusions: The present study demonstrates the association between the SPTLC3 rs364585 SNP and serum TC, HDL-C and LDL-C levels in our study populations. These associations might have ethnic- and/or sex-specificity. Trial registration: Retrospectively registered. [ABSTRACT FROM AUTHOR]

The article discusses clinical characteristics of patients having alpha-1-antitrypsin (A1AT), protease inhibitor (serpins). Topics discussed include controlling of coagulation, usage of an optical aggregometer, and separation of globulin by agarose gel electrophoresis. Particular focus is also given to approval of the study by the board of research ethic at Jiangsu Institute of Haematology in China.

Rice straw (RS) is an important raw material for the preparation of Agaricus bisporus compost in China. In this study, the characterization of composting process from RS and wheat straw (WS) was compared for mushroom production. The results showed that the temperature in RS compost increased rapidly compared with WS compost, and the carbon (C)/nitrogen (N) ratio decreased quickly. The microbial changes during the Phase I and Phase II composting process were monitored using denaturing gradient gel electrophoresis (DGGE) and phospholipid fatty acid (PLFA) analysis. Bacteria were the dominant species during the process of composting and the bacterial community structure dramatically changed during heap composting according to the DGGE results. The bacterial community diversity of RS compost was abundant compared with WS compost at stages 4-5, but no distinct difference was observed after the controlled tunnel Phase II process. The total amount of PLFAs of RS compost, as an indicator of microbial biomass, was higher than that of WS. Clustering by DGGE and principal component analysis of the PLFA compositions revealed that there were differences in both the microbial population and community structure between RS- and WS-based composts. Our data indicated that composting of RS resulted in improved degradation and assimilation of breakdown products by A. bisporus, and suggested that the RS compost was effective for sustaining A. bisporus mushroom growth as well as conventional WS compost. [ABSTRACT FROM AUTHOR]

Within-lake heterogeneity of bacterial community composition (BCC) was investigated in the large and shallow regulating Lake Dongting. Samples were collected at 13 sites located in different areas of the lake. PCR-denaturing gradient gel electrophoresis (DGGE) and redundancy analysis (RDA) were used for revealing spatial distribution of BCC and the relationships between BCC and environmental variables. The DGGE banding patterns revealed a remarkable spatial heterogeneity which was closely related to their geographical positions. RDA result demonstrated that TP and TN, as well as Secchi depth, were the three most influential factors, responsible for a major part of the observed variation in BCC. Total bacterial abundances were significantly higher in Eastern Dongting due to high TP and suspended solids. In conclusion, bacterial community diversity in Lake Dongting was mainly shaped by within-lake heterogeneity of nutrient and transparency. [ABSTRACT FROM AUTHOR]

Filamentous Nostoc flagelliforme form colloidal complex, with beaded cells interacting with other bacteria embedded in the complex multilayer sheath. However, the species of bacteria in the sheath and the interaction between N. flagelliforme and associated bacteria remain unclear. In this study, PCR-denaturing gradient gel electrophoresis (DGGE) was used to investigate the bacterial communities of N. flagelliforme from three regions of China. DGGE patterns showed variations in all samples, exhibiting 25 discrete bands with various intensities. The diversity index analysis of bands profiles suggested the high similarity of bacterial communities to each other but also the dependence of microbial composition on each location. Phylogenetic affiliation indicated that the majority of the sequences obtained were affiliated with Actinobacteria, Cyanobacteria, Proteobacteria, Acidobacteria, Bacteroidetes, of which Cyanobacteria was dominant, followed the Proteobacteria. Members of the genus Nostoc were the most abundant in all samples. Rhizobiales and Actinobacteria were identified, whereas, Craurococcus, Caulobacter, Pseudomonas, Terriglobus and Mucilaginibacter were also identified at low levels. Through comparing the bacterial composition of N. flagelliforme from different regions, it was revealed that N. flagelliforme could facilitate the growth of other microorganisms including both autotrophic bacteria and heterotrophic ones and positively contributed to their harsh ecosystems. The results indicated N. flagelliforme played an important role in diversifying the microbial community composition and had potential application in soil desertification. [ABSTRACT FROM AUTHOR]

Sediment cores taken from different areas of the mesotrophic Lake Erhai were analysed to investigate the vertical distribution of bacterial community composition (BCC), as well as physicochemical parameters. PCR-denaturing gradient gel electrophoresis (DGGE), stable carbon isotope (δ13C), C/N atomic ratio and canonical correspondence analysis (CCA) were used to explore the relationships between the succession of bacterial communities and environmental variables, emphasising changes in the sources of organic matter (OM). The BCC in natural environments was characterised by DGGE of the 16S rRNA gene with subsequent sequencing of bands of interest. The CCA revealed that the depth-related variation in sediment bacterial communities in different areas of the lake was significantly influenced by varying environmental factors. The OM source, however, played an important role in structuring BCC at all sites. The DGGE banding patterns revealed that the abundance of Deltaproteobacteria decreased with accompanying elevated levels of C4 plant-derived organic carbon. The sequencing of DGGE bands suggested that the majority of the sequences were affiliated with common phylogenetic groups in lake sediments: Chloroflexi, Deltaproteobacteria and Firmicutes. Betaproteobacteria detected in our study appeared as a prominent phylotype in the upper sediment. The Shannon-Wiener diversity index of bacterial communities was directly affected by the OM source. Constant OM sources resulted in a stable higher diversity of bacterial communities and broader enzymatic capabilities to access OM. We conclude that the differences in the diversity of bacterial communities in sediments differing in their sources of OM were related to environmental variables (e.g. water level, river runoff and terrestrial vegetation composition). Our study provided insights into the relationships between natural BCC and OM sources, facilitating a better understanding of microbial community structure in lake sediment. [ABSTRACT FROM AUTHOR]

Effects of oxygen inoculation were investigated on oil biodegradation and biodiversity of reservoir microorganism in Dagang oil field. The biodegrading was evaluated and analyzed or estimated by using in-vitro enrichment, denaturing gradient gel electrophoresis, real time PCR and GC–MS. The bacterial communities of production water containing reservoir microorganism were 30% lower than groundwater and injection water despite that all of them have rich biodiversity. After thirty days of oxygen treatment the n -alkanes degradation were 92.5% in injected water while 44.3% in groundwater and 92.4% in production water. The selected PAHs biodegradation rate in production water was lower as compared to the injection water but significantly higher than groundwater. The total bacterial biomass gradually increases after the oxygen injection and then decreases. The results suggested that aerobic condition could accelerate petroleum biodegradation and changes the microbial community structure in oil reservoir, which might be beneficial to MEOR. [ABSTRACT FROM AUTHOR]

Objective To collect the radioactive contamination data for environmental rehabilitation in uranium mill tailings in southeastern China. Method The sample areas were divided into high, moderate and low concentration areas, according to the uranium concentration. For every area, 3 soil samples were collected at 0–15 cm, 15–30 cm and 30–45 cm depth respectively, with 5 repetitions for each. Total 45 (3 × 5 × 3) soil samples were collected. Physicochemical properties and enzyme activities of soils were determined as described by references. The concentrations of the radionuclides 238 U, 232 Th, 226 Ra and 40 K in soils were determined by using HPGe gamma-ray spectrometer. Soil microbial diversity was analyzed via denaturing gradient gel electrophoresis (DGGE). Results Soil samples were all acidic. Physicochemical properties, like pH, content of total/available N, P and K, as well as enzyme activities were all increased along with decreased uranium concentration. The 232 Th concentration was increased with the decreased uranium concentration and was not influenced by the depth of sample sites. However, uranium concentration and depth of sample showed no significant influence on the concentrations of 226 Ra and 40 K. The concentration of 232 Th was significantly correlated with that of 226 Ra and 40 K, while the concentrations of 226 Ra and 40 K were significantly correlated. However, Pearson correlation coefficients between 238 U and other radionuclides were not significant. The microbial population in different concentration areas was different with four domain strains in low area, and two for both moderate and high areas. Furthermore, in each sample site, Proteobacteria was the most dominant flora, while environmental samples were the second according to GenBank database. Moreover, Serratia sp. of Proteobacteria was the dominant strain. Conclusion Radionuclides distribution in the uranium mill tailing showed a profound influence on soil properties and microbial diversity. This primarily study might provide valuable data for further research towards a better understanding of the radioactive contamination in uranium mill tailings in southeast China. [ABSTRACT FROM AUTHOR]

To clarify the relationships between dissolved organic carbon (DOC) and bacterioplankton community composition (BCC), a 1-year survey (June 2009 - May 2010) was conducted in 3 regions of Lake Taihu (Meiliang Bay, Lake Center, and Eastern Taihu), China. Polymerase chain reaction - denaturing gradient gel electrophoresis was used to analyze the composition and heterogeneity of the bacterioplankton community. Canonical correspondence analysis was used to explore the relationships between DOC concentration and BCC. We found a significant negative correlation between DOC concentration and bacterioplankton community diversity (as measured by the Shannon-Wiener index ( H′)). The results show that spatial variation in the bacterioplankton population was stronger than the seasonal variation and that DOC concentration influences BCC in Lake Taihu. DOC concentration, followed by macrophyte biomass, water turbidity, and phytoplankton biomass were the most influential factors that account for BCC changes in Lake Taihu. More detailed studies on the relationship between DOC concentration and BCC should focus on differences in DOC concentrations and quality among these lake regions. DOC had a significant impact on BCC in Meiliang Bay. The relationship between DOC and BCC in the 2 other regions studied (Lake Center and Eastern Taihu) was weaker. The results of this study add to our understanding of the BCC in eutrophic lakes, especially regarding the role of the microbial loop in lake ecosystems. [ABSTRACT FROM AUTHOR]

Background: Under the existing national surveillance system in China for selected infectious diseases, bacterial cultures are performed for only a small percentage of reported cases. We set up a laboratory-based syndromic surveillance system to elucidate bacterial etiologic spectrum and detect infection by rare etiologies (or serogroups) for five core syndromes in the given study area. Methods: Patients presenting with one of five core syndromes at nine sentinel hospitals in Guagnxi, China were evaluated using laboratory-based syndrome surveillance to elucidate bacterial etiologies. We collected respiratory and stool specimens, as well as CSF, blood and other related samples for bacterial cultures and pulse field gel electrophoresis (PFGE) assays. Results: From February 2009 to December 2011, 2,964 patients were enrolled in the study. Etiologies were identified in 320 (10.08%) patients. Streptococcus pneumonia (37 strains, 24.18%), Klebsiella pneumonia (34, 22.22%), Pseudomonas aeruginosa (19, 12.42%) and Haemophilus influenza (18, 11.76%) were the most frequent pathogens for fever and respiratory syndrome, while Salmonella (77, 81.05%) was most often seen in diarrhea syndrome cases. Salmonella paratyphi A (38, 86.36%) occurred in fever and rash syndrome, with Cryptococcus neoformans (20, 35.09%), Streptococcus pneumonia (5, 8.77%), Klebsiella pneumonia (5, 8.77%),streptococcus suis (3, 5.26%) and Neisseria meningitides group B (2, 3.51%) being the most frequently detected in encephalitis-meningitis syndrome. To date no pathogen was isolated from the specimens from fever and hemorrhage patients. Conclusions: In addition to common bacterial pathogens, opportunistic pathogens and fungal infections require more attention. Our study contributes to the strengthening of the existing national surveillance system and provides references for other regions that are similar to the study area. [ABSTRACT FROM AUTHOR]

Hyperthermostable proteins are highly resistant to various extreme conditions. Many factors have been proposed to contribute to their ultrahigh structural stability. Some thermostable proteins have larger oligomeric size when compared to their mesophilic homologues. The formation of compact oligomers can minimize the solvent accessible surface area and increase the changes of Gibbs free energy for unfolding. Similar to mesophilic proteins, hyperthermostable proteins also face the problem of unproductive aggregation. In this research, we investigated the role of high-order oligomerization in the fight against aggregation by a hyperthermostable superoxide dismutase identified from Tengchong, China (tcSOD). Besides the predominant tetramers, tcSOD could also form active high-order oligomers containing at least eight subunits. The dynamic equilibrium between tetramers and high-order oligomers was not significantly affected by pH, salt concentration or moderate temperature. The secondary and tertiary structures of tcSOD remained unchanged during heating, while cross-linking experiments showed that there were conformational changes or structural fluctuations at high temperatures. Mutational analysis indicated that the last helix at the C-terminus was involved in the formation of high-order oligomers, probably via domain swapping. Based on these results, we proposed that the reversible conversion between the active tetramers and high-order oligomers might provide a buffering system for tcSOD to fight against the irreversible protein aggregation pathway. The formation of active high-order oligomers not only increases the energy barrier between the native state and unfolded/aggregated state, but also provides the enzyme the ability to reproduce the predominant oligomers from the active high-order oligomers. [ABSTRACT FROM AUTHOR]

Previous studies on microbial prospecting of oil/gas only focused on the anomalies of light hydrocarbon-oxidizing microbes as main exploratory indicators and their exploration applications. In this study, we investigated the responses of microbial communities to light-hydrocarbon microseepage in the Beihanzhuang Oilfield, eastern China using denaturing gradient gel electrophoresis (DGGE) analysis and by comparing the difference of two-type areas with high- and low-flux light-hydrocarbon seepages. The results showed that the high-flux light-hydrocarbon seepage favored the growth ofNocardioides,Aciditerrimonas, sulphate-reducing bacteria (SRBs) related toDesulfosporosinusandDesulfovibrio, and Chloroflexi bacteria (b-7), implying that their anomalies might be adopted as novel subsidiary indicators for microbial prospecting of oil/gas in the Beihanzhuang Oilfield. Based on the newly obtained results, we have proposed a general strategy for microbial prospecting of oil/gas, i.e., to determine the anomalies of light hydrocarbon-oxidizing microbes, to select subsidiary indicators for microbial prospecting of oil/gas based on an assessment of the responses of microbial communities to light-hydrocarbon microseepage, to quantitatively measure subsidiary indicators and delimit their anomalies, to comprehensively interpret all microbial anomalies, and to make a suggestion for oil/gas prospecting. This general strategy with novel indicators may provide a more comprehensive evaluation for light-hydrocarbon microseepage and the corresponding anomalies, thereby reducing the exploration risk of oil/gas. [ABSTRACT FROM AUTHOR]

A total of 310 Salmonella isolates were isolated from 6 broiler farms in Eastern China, serotyped according to the Kauffmann-White classification. All isolates were examined for susceptibility to 17 commonly used antimicrobial agents, representative isolates were examined for resistance genes and class I integrons using PCR technology. Clonality was determined by pulsed-field gel electrophoresis (PFGE). There were two serotypes detected in the 310 Salmonella strains, which included 133 Salmonella enterica serovar Indiana isolates and 177 Salmonella enterica serovar Enteritidis isolates. Antimicrobial sensitivity results showed that the isolates were generally resistant to sulfamethoxazole, ampicillin, tetracycline, doxycycline and trimethoprim, and 95% of the isolates sensitive to amikacin and polymyxin. Among all Salmonella enterica serovar Indiana isolates, 108 (81.2%) possessed the blaTEM, floR, tetA, strA and aac (6')-Ib-cr resistance genes. The detected carriage rate of class 1 integrons was 66.5% (206/310), with 6 strains carrying gene integron cassette dfr17-aadA5. The increasing frequency of multidrug resistance rate in Salmonella was associated with increasing prevalence of int1 genes (rs = 0.938, P = 0.00039). The int1, blaTEM, floR, tetA, strA and aac (6')-Ib-cr positive Salmonella enterica serovar Indiana isolates showed five major patterns as determined by PFGE. Most isolates exhibited the common PFGE patterns found from the chicken farms, suggesting that many multidrug-resistant isolates of Salmonella enterica serovar Indiana prevailed in these sources. Some isolates with similar antimicrobial resistance patterns represented a variety of Salmonella enterica serovar Indiana genotypes, and were derived from a different clone. [ABSTRACT FROM AUTHOR]

Background Shiga toxin-producing Escherichia coli (STEC) is recognized as an important human diarrheal pathogen. Swine plays an important role as a carrier of this pathogen. In this study we determined the prevalence and characteristics of STEC from healthy swine collected between May 2011 and August 2012 from 3 cities/provinces in China. Results A total of 1003 samples, including 326 fecal, 351 small intestinal contents and 326 colon contents samples, was analyzed. Two hundred and fifty five samples were stx-positive by PCR and 93 STEC isolates were recovered from 62 stx-positive samples. Twelve O serogroups and 19 O:H serotypes including 6 serotypes (O100:H20/[H20], O143:H38/[H38], O87:H10, O172:H30/[H30], O159:H16, O9:H30/[H30]) rarely found in swine and ruminants were identified. All 93 STEC isolates harbored stx2 only, all of which were stx2e subtype including 1 isolate being a new variant of stx2e. 53.76%, 15.05% and 2.15% STEC isolates carried astA, hlyA and ehxA respectively. Four STEC isolates harbored the high-pathogenicity island. Of the 15 adherence-associated genes tested, 13 (eae, efa1, iha, lpfAO113, lpfAO157/OI- 154, lpfAO157/OI-141, toxB, saa, F4, F5, F6, F17 or F41) were all absent while 2 (paa and F18) were present in 7 and 4 STEC isolates respectively. The majority of the isolates were resistant to tetracycline (79.57%), nalidixic acid (78.49%), trimethoprim-sulfamethoxazole (73.12%) and kanamycin (55.91%). The STEC isolates were divided into 63 pulsed-field gel electrophoresis patterns and 21 sequence types (STs). Isolates of the same STs generally showed the same or similar drug resistance patterns. A higher proportion of STEC isolates from Chongqing showed multidrug resistance with one ST (ST3628) resistant to 14 antimicrobials. Conclusions Our results indicate that swine is a significant reservoir of STEC strains in China. Based on comparison by serotypes and sequence types with human strains and presence of virulence genes, the swine STEC may have a low potential to cause human disease. [ABSTRACT FROM AUTHOR]

Abstract: Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae (GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the first time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in five provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding 36.9% (62/168), 37.5% (63/168) and 19.6% (33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia (95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010, but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during 2007–2011, III only occurred in 2012. 14 different PFGE types, including 4 new types (N, O, P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9% (141/168) of the total GBS strains. Ia corresponded to 11 PFGE types (A–H, N–P), in which type D predominated (51%). Ib represented 3 genotypes (I, J and Q) and III harbored only 2 genotypes (N and F). Type N and F synchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years. Although Ia still predominated, new rare serotype III already occurred in China. [Copyright &y& Elsevier]

Abstract: Objective: To investigate molecular characterization of streptococcus pyogenes isolates involved in an outbreak of scarlet fever in China in 2011. Methods: Seventy-four Streptococcal pyogenes involved in an outbreak of scarlet fever were isolated from pediatric patients in the areas with high incidence in China from May to August of 2011. Emm genotyping, pulsed-field gel electrophoresis (PFGE), superantigen (SAg) genes and antimicrobial susceptibility profiling were analyzed for these isolates. Results: A total of 4 different emm types were identified. Emm12 was the most prevalent type which contained four predominating PFGE patterns corresponding to four different virulence and superantigen profiles. Emm12 (79.7%) and emm1 (14.9%) accounted for approximately 94% of all the isolates. The speA gene was all negative in emm12 isolates and positive in emm1 isolates. All strains were resistant to erythromycin, and 89.4% of them were resistant to erythromycin, tracycline, and clindamycin simultaneously. Conclusion: Several highly diversified clones with a high macrolide resistance rate comprise a predominant proportion of circulating strains, though no new emm type was found in this outbreak. The data provide a baseline for further surveillance of scarlet fever, which may contribute to the explanation of the outbreak and development of a GAS vaccine in China. [Copyright &y& Elsevier]

Indigenous Mycobacterium communities play an important role in the degradation of polycyclic aromatic hydrocarbons (PAHs), but little is known about Mycobacterium distribution in situ at PAH-contaminated sites. In this study, the diversity and distribution of Mycobacterium communities were investigated in sediments and soils at sites upstream, midstream, and downstream of an oil-sewage irrigation channel, using denaturing gradient gel electrophoresis (DGGE). The results show that heavy PAH contamination in upstream sites negatively affected Mycobacterium community diversity compared with midstream and downstream sites in all 3 sample types (sediments, corn field soils, and rice field soils). There was a correlation between the distribution of Mycobacterium communities and PAH contamination, as indicated by canonical correspondence analysis. Mycobacterium diversity and distribution was found to vary between the 3 sample types. [ABSTRACT FROM AUTHOR]

Objectives: To define the characteristic of bacteremia in patients with acute pancreatitis and determine its possible association with the disease severity. Design: A prospective controlled study. Setting: ICU of Jinling Hospital, China. Patients: A total of 48 patients with mild or severe acute pancreatitis were enrolled in the study. Interventions: None. Measurements and Main Results: Samples of peripheral blood were collected from the patients at 4 or 5 and 9 or 10 days after acute pancreatitis was definitely diagnosed. Resulting DNA from the blood was analyzed using denaturing gradient gel electrophoresis, and separated fragments were sequenced for identification of bacterial species. Bacterial DNA was detected in peripheral blood from 68.8% of patients with acute pancreatitis, and more than half (60.4%) of the patients encountered polymicrobial flora. Translocated bacteria ri patients with acute paricreatitis were primarily constituted of opportunistic pathogens derived from the gut, including Escherichia coli, Shigella ulexneri, Enterobacteriaceae bacterium, Acinetobacter Iwoffii, Bacillus coagulans, and Enterococcus faecium. The species of circulating bacteria shifted remarkably among the patients with different severity. The presence of the bacteremia correlated positively with the Acute Physiology and Chronic Health Evaluation-II scores of patients with acute paricreatitis (r = 0.7918, p < 0.0001). Conclusions: This study provides a detailed description on the prevalence of bacteremia and characteristic of bacterial species in patients with acute pancreatitis. We demonstrate an association between the bacteremia and the disease severity, which enables us to better understand a potential role of bacterial translocation in the pathogenesis of septic complication in acute pancreatitis. [ABSTRACT FROM AUTHOR]

Much more attention has been paid to the actinobacterial community in soils or water columns of aquatic habitats. However, there are few studies on their composition and diversity in lake sediments. Here, we used denaturing gradient gel electrophoresis and clone libraries of partial 16S rRNA gene to study the spatial variations of actinobacterial communities across 4 seasons in the surface sediments of the shallow, subtropical Taihu Lake. Cluster analysis based on fingerprints showed clear spatiotemporal variations of actinobacterial communities and higher seasonal variation than spatial heterogeneity. Based on clone libraries, this pattern was supported by the principal coordinates analysis in the phylogenetic context and by detrended correspondence analysis on the operational taxonomic unit table. Additionally, phylogenetic analysis showed that the putative freshwater-specific actinobacterial lineages (e.g., acI) were also detected in the lake sediments, which suggests that these subclusters may also adapt to the sediment environments. Summarily, our results suggested that actinobacterial communities of the surface sediments were more affected by seasonal variation than spatial heterogeneity in the intrahabitat of Taihu Lake. [ABSTRACT FROM AUTHOR]

This study investigated effects of wet and dry seasons on the bacterial community structure of the Three Gorges Reservoir by using denaturing gradient gel electrophoresis analysis of the PCR-amplified bacterial 16S rRNA gene. Bacterial diversity, as determined by the Shannon index, the Simpson's index, and the Richness, dramatically changed in between the dry and wet seasons. The changes in the diversity and relative abundance of microbial populations among the five sites during the wet season have become more marked than those observed during the dry season. Furthermore, cluster analysis also showed these changes. The phylogenetic analysis indicated that Betaproteobacteria is the dominant population, followed by Actinobacterium, in both the wet season and dry season. The water quality parameters were quite stable at all five sites during the same season but noticeably varied from season to season. Canonical correspondence analysis also indicated that the changes in the bacterial community composition were primarily correlated with the variations in temperature, transparency, and the concentrations of NH-N. Slight changes in bacterial community composition among the five sites during the dry season were correlated with different environments. However, during the wet season, major changes were correlated not only with environments, but also it may be associated with the bacterial populations from the surrounding areas and tributaries of the Three Gorges Reservoir. [ABSTRACT FROM AUTHOR]

To identify the methanogenic pathways present in a deep coal bed methane (CBM) reservoir associated with Eastern Ordos Basin in China, a series of geochemical and microbiological studies was performed using gas and water samples produced from the Liulin CBM reservoir. The composition and stable isotopic ratios of CBM implied a mixed biogenic and thermogenic origin of the methane. Archaeal 16S rRNA gene analysis revealed the dominance of the methylotrophic methanogen Methanolobus in the water produced. The high potential of methane production by methylotrophic methanogens was found in the enrichments using the water samples amended with methanol and incubated at 25 and 35 °C. Methylotrophic methanogens were the dominant archaea in both enrichments as shown by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE). Bacterial 16S rRNA gene analysis revealed that fermentative, sulfate-reducing, and nitrate-reducing bacteria inhabiting the water produced were a factor in coal biodegradation to fuel methanogens. These results suggested that past and ongoing biodegradation of coal by methylotrophic methanogens and syntrophic bacteria, as well as thermogenic CBM production, contributed to the Liulin CBM reserves associated with the Eastern Ordos Basin. [ABSTRACT FROM AUTHOR]

The diversity of buoyant Microcystis colony-associated, settling particle-associated and free-living freshwater bacteria in the eutrophic Lake Taihu in China was compared by the polymerase chain reaction-denaturing gradient gel electrophoresis ( PCR-DGGE) of 16S rRNA genes and by clone library analysis. The cluster analysis of DGGE profiles revealed that the buoyant Microcystis colony associates collected in summer and winter were clustered together and were distinct from settling particle-associated and free-living bacteria. In contrast, the bacterial communities of the latter two populations collected in summer were clustered together and varied from those collected in winter. The diversity indices of the Microcystis-associated bacterial population were significantly lower than those of the other two bulk bacterial communities ( P < 0.05). Clone library analysis revealed that no shared operational taxonomic units were found in the three populations. Phylogenetic analysis confirmed that buoyant Microcystis-associated bacteria were distinct from the other communities and that they were dominated by Alphaproteobacteria, Betaproteobacteria and Gammaproteobacteria. Proteobacteria, Bacteroidetes and Actinobacteria were prevalent in all three habitats. Other bacterial taxa such as Planctomycetes, Verrucomicrobia, Gemmatimonadetes and Acidobacteria were associated with settling particles and with the water column. Buoyant Microcystis colonies, settling particles and the water column in the eutrophic lake are thus inhabited by different bacterial flora. In addition, specific bacterial communities are associated with buoyant Microcystis colonies. [ABSTRACT FROM AUTHOR]

Clin Microbiol Infect Abstract The sporadic emergence of New Delhi metallo-β-lactamase-1 (NDM-1)-producing Acinetobacter spp. has been reported in China; however, NDM-1-positive bacteria epidemics are rarely reported in intensive care units (ICUs) in China, or even in the world. During 15 months' surveillance Acinetobacter spp. isolated from patients, heathcare workers and surfaces of a Chinese ICU were screened for the blaNDM-1 gene. A total of 27 of 3114 Acinetobacter spp. strains were NDM-1 positive and identified as A. pittii with sequence type 63 (ST63) by multilocus sequence typing. Of the 27 NDM-1-positive A. pittii strains, 22 were isolated from the ICU surfaces and grouped into a major clone A using pulsed-field gel electrophoresis typing, while the other five strains isolated from the patients were classified into three clones (A, B and C). The blaNDM-1 gene was located on a 45-kb plasmid for all three A. pittii clones. The plasmid could be transferred to A. pittii and A. baumannii recipients at both 30 and 37°C but not to Escherichia coli J53. The plasmid could not be classified into any of the known plasmid incompatibility groups. The blaNDM-1 region in the plasmid was flanked by two insertion sequence elements, IS Aba125 and IS Aba11, and no other carbapenemase gene was present in this NDM-1-positive A. pittii isolate. Thus, we present the first report on the transmission and characterization of NDM-1-producing A. pittii in an ICU in China as well as a novel blaNDM-1 gene-bearing plasmid. [ABSTRACT FROM AUTHOR]

The nature of microbial communities and their relation to enzyme activities in desert soils is a neglected area of investigation. To address this, the bacterial diversity and distribution and soil physico-chemical factors were investigated in the soil crust, the soil beneath the crust and rhizosphere soil at the southeast edge of the Tengger Desert, using the denaturing gradient gel electrophoresis of 16S rRNA genes amplified by the polymerase chain reaction. Phylogenetic analysis of the sequenced DGGE bands revealed a great diversity of bacteria. The Proteobacteria, consisting of the ɑ, β, and γ subdivisions, were clearly the dominant group at all depths and in rhizosphere soil. Analysis of the enzyme activities indicated that the rhizosphere soil of Caragana korshinskii exhibited the highest protease and polyphenol oxidase activities, and in the soil crust there were increased activities of catalase, urease, dehydrogenase and sucrase. The bacterial community abundance closely correlated with soil enzyme activities in different soils. The presence of Cyanobacteria correlated with significant increases in protease, catalase and sucrase in the soil crust, and increased urease in the rhizosphere soil of Artemisia ordosica. The occurrence of Acidobacteria was associated with significant increases in urease, dehydrogenase, and sucrase in the rhizosphere soil of C. korshinski. The presence of γ-Proteobacteria correlated with a significant increase in polyphenol oxidase in the rhizosphere soil of A. ordosica. The study indicated a close relationship between the soil bacterial community and soil enzymes, suggesting the necessity of further investigations into bacterial function in this desert ecosystem. [ABSTRACT FROM AUTHOR]

Abstract: Large-scale streptococcal outbreaks occurred continuously in tilapia farms of China from 2009 to 2011. The objective of this study was to characterize the prevalent strains of tilapia streptococci from the main cultured areas of China through species specific PCR and pulse field gel electrophoresis (PFGE). A total of 105 prevalent strains were isolated from Guangdong, Guangxi, Hainan and Fujian provinces between 2006 and 2011, 85 of which were identified as Streptococcus agalactiae while the rest were all identified as Streptococcus iniae. The prevalent stains in 2006 and 2007 were S. iniae (94.7%, 18/19), with S. agalactiae account for only 5.3% (1/19); The prevalent strains in 2009 and 2011 however changed to S. agalactiae (97.7%, 84/86), with only 2.3% (2/86) was S. iniae. Of these 105 strains, a total of 13 PFGE types (A-M) were characterized, among which D, F, G and K genotypes were predominant, accounting for 81.90% (86/105). The cluster analysis of PFGE electropherograms separated S. iniae and S. agalactiae to two distinctive branches, 20 strains of S. iniae exhibiting 3 types of PFGE band patterns with a similarity of 94.8–100%, and the 85 strains of S. agalactiae producing 10 types of PFGE band patterns with a similarity between 48.4% and 100%. Data suggested that the prevalent strains of tilapia streptococci in China have shifted from the former (before 2008) dominant strains of S. iniae to the current (2009–2011) dominant strains of S. agalactiae. Moreover, PFGE genotypes of the prevalent strains demonstrated geographic differences and temporal changes. [Copyright &y& Elsevier]

Abstract: Laribacter hongkongensis is a novel emerging pathogen associated with human gastroenteritis. We aimed to investigate the prevalence, antimicrobial resistance and genotypic relationship of 199 L. hongkongensis isolates from 690 intestinal samples of fish and frogs. These samples were collected from retail markets in the city of Guangzhou in southern China from October 2008 to September 2009. L. hongkongensis was detected in from 80 (16.3%) out of 490 freshwater fish, and this number included 76 (32.3%) out of 235 grass carp and 4 (14.8%) out of 27 bighead carp. A higher isolation rate of 59.5% (119 out of 200) was observed in edible frogs. The isolation rate was highest in the spring in comparison with other seasons. Notably, 63.8% of the isolates were resistant to at least one class of antimicrobial agents. Analysis by pulsed-field gel electrophoresis (PFGE) revealed that the isolates could be grouped into three clusters. Isolates from fish intestines were grouped into two clusters: cluster I and II. Isolates of frog-origin and several fish-origin isolates were grouped into cluster III. Two patient-derived strains could be classed into cluster III. Extensive genetic heterogeneity among the isolates was observed. The results indicate that L. hongkongensis isolates exhibits host tropism, extensive resistance to widely used antimicrobials and diverse biological evolution in an aquatic environment. The frog is more likely than the freshwater fish to be the potential source for human infection with L. hongkongensis. [Copyright &y& Elsevier]

Significant spatial variation in CH emissions is a well-established feature of natural wetland ecosystems. To understand the key factors affecting CH production, the variation in community structure of methanogenic archaea, in relation to substrate and external environmental influences, was investigated in selected wetlands across China, using denaturing gradient gel electrophoresis. Case study areas were the subtropical Poyang wetland, the warm-temperate Hongze wetland, the cold-temperate Sanjiang marshes, and the alpine Ruoergai peatland on the Qinghai-Tibetan Plateau. The topsoil layer in the Hongze wetland exhibited the highest population of methanogens; the lowest was found in the Poyang wetland. Maximum CH production occurred in the topsoil layer of the Sanjiang Carex lasiocarpa marsh, the minimum was observed in the Ruoergai peatland. CH production potential was significantly correlated with the dissolved organic carbon (DOC) concentration but not with the abundance or diversity indices of methanogenic archaea. Phylogenetic analysis and DOC concentration indicated a shift in the dominant methanogen from the hydrogenotrophic Methanobacteriales in DOC-rich wetlands to Methanosarcinaceae with a low affinity in wetlands with relatively high DOC and then to the acetotrophic methanogen Methanosaetaceae with a high affinity in wetlands with low DOC, or with high DOC but rich sulfate-reducing bacteria. Therefore, it is proposed that the dominant methanogen type in wetlands is primarily influenced by available DOC concentration. In turn, the variation in CH production potential in the wetlands of eastern China is attributable to differences in the DOC content and the dominant type of methanogen present. [ABSTRACT FROM AUTHOR]