In vitro Antibiotic Susceptibility, Virulence Genes Distribution and Biofilm Production of Staphylococcus aureus Isolates from Bovine Mastitis in the Liaoning Province of China.

PURPOSE: The aim of this study was to identify the subtype, characterize the antimicrobial resistance, determine the virulence gene distribution, and analyze the biofilm production of Staphylococcus aureus isolates from bovine mastitis milk samples in the Liaoning Province of China. MATERIALS AND METHODS: In total, 56 Staph. aureus isolates were collected and identified in this study; the isolates were divided into different spa types based on the sequence of the polymorphic X region of the spa gene. Additionally, antimicrobial susceptibility was investigated using the broth microdilution method, and 18 virulence genes were detected using PCR. Biofilm formation was measured by spectrophotometry with crystal violet staining and observed using confocal laser scanning microscopy. RESULTS: There were 12.12% (56/462) milk samples that were positive for Staph. aureus. These isolates were nonsusceptible to sulfamethoxazole (100%), penicillin (76.9%), daptomycin (76.79%), clindamycin (69.64%), and oxacillin (60.71%); however, the majority of the isolates (80.4%) were susceptible to amoxicillin/clavulanate. The predominant virulence genes encoded the cytotoxins, hla (94.64%) and hlb (89.29%), and the adhesion factors clfA (89.29%), clfB (89.29%), and fnbB (80.36%). Comparatively, virulence genes related to other adhesion factors such as cna (8.93%) and enterotoxins, such as seg (26.79%), sea (16.07%), seb (7.14%), and sec (7.14%) were detected at relatively lower rates. The following eight spa types were identified: t267 (35.84%), t730 (22.64%), t518 (15.09%), t1190 (11.32%), t1456 (9.43%), t224 (1.88%), t9129 (1.88%), and t177 (1.88%). The highest biofilm production was observed for t267. Staph. aureus exhibited various patterns of biofilm formation, with the biofilm often being associated with a tower-shaped structure or a thicker biofilm. CONCLUSION: Our results indicated that Staph. aureus isolates from dairy cows with mastitis in the Liaoning Province of China were non-susceptible to sulfamethoxazole, penicillin, daptomycin, oxacillin, and clindamycin. Additionally, the most prevalent subtype was t267, which displayed resistance to multiple antimicrobial agents and harbored several virulence genes, including clfA, clfB, fnbB, hla, and hlb.

[Pilot-scale Experiment on Enrichment of Nitrifying Activated Sludge and Its Application in Enhancing a Wastewater Biological Treatment System Against Ammonia Shocking Loads].

Nitrifying activated sludge (NAS) was enriched in a membrane bioreactor (MBR) with pre-treated municipal wastewater and additional ammonium sulfate as the culture medium. The influences of temperature, dissolved oxygen (DO), ammonia nitrogen volumetric load, free ammonia (FA), and free nitrite (FNA) on the enrichment of NAS were investigated, the cost of the process was evaluated, and then NAS's application in enhancing a wastewater biological treatment system against ammonia shocking loads was attempted. The results showed that after 182 days of cultivation in an MBR, NAS had a nitrification activity of 98.41 mg·(L·h)-1, which was 30-times higher than that of the seeding sludge. The yield of NAS was 14.96 mg·(L·d)-1, costing 3.52 Yuan for 1 kg. Temperature was found to be a key factor affecting the sludge nitrification activity. The sludge nitrification activity was decreased to 1/3 of the maximum value at temperatures below 15.0℃, while lowering the ammonium volumetric load retarded the decrease in the sludge nitrification activity to some extent. In addition, dissolved oxygen deficiency resulted in nitrite accumulation, and thereby slowed down the NAS enrichment rate. The enriched NAS was then applied to a wastewater biological treatment pilot equipment, which had just been exposed to an ammonium shocking load. The removal rate of ammonia nitrogen in the biological system increased from 29.4% to 88.4% after 2.0% of NAS was inoculated. The enhanced biological system retained ammonia removal rates of as high as 99.0%, even as the temperature dropped to 13.3℃±1.6℃ afterwards. The above pilot-experiment results suggested that enriched nitrifying sludge is suitable for quickly increasing the start-up or recovery rates of the nitrifying function in a biological system.

Association of polyunsaturated fatty acids in breast milk with fatty acid desaturase gene polymorphisms among Chinese lactating mothers.

BACKGROUND: The fatty acid desaturase (FADS) controls polyunsaturated fatty acid (PUFA) synthesis in human tissues and breast milk. DESIGN: Evaluate the influence of 10 single nucleotide polymorphisms (SNPs) and various haplotypes in the FADS gene cluster (FADS1, FADS2, FADS3) on PUFA concentration in the breast milk of 209 healthy Chinese women. PUFA concentrations were measured in breast milk using gas chromatography and genotyping was performed using the Sequenom Mass Array system. RESULTS: A SNP (rs1535) and 2-locus haplotypes (rs3834458-rs1535, rs1535-rs174575) in the FADS2 gene were associated with concentrations of γ-linoleic acid (GLA) and arachidonic acid (AA) in breast milk. Likewise, in the FADS1 gene, a 2-locus constructed haplotype (rs174547-rs174553) also affected GLA and AA concentration (P<0.05 for all). Minor allele carriers of the SNP and haplotypes described above had lower concentrations of GLA and AA. In the FADS2 gene, the 3-locus haplotype rs3834458-rs1535-rs174575, significantly affected concentrations of GLA but not AA. Pairwise comparison showed that individuals major homozygous for the SNP rs1000778 in the FADS3 gene had lower concentrations of ALA and linoleic acid (LA) in their breast milk. CONCLUSION: Polymorphisms in the FADS gene cluster influence PUFA concentrations in the breast milk of Chinese Han lactating women.