Author: Barbosa, Kristian da Silva ; Pereira, Ligia Beatriz Rizzanti ; Dos Santos, Marcos Veiga ; Garcia, Breno Luis Nery ; Dantas, Stéfani Thais Alves ; Fidelis, Carlos Eduardo ; Rall, Vera Lúcia Mores

Streptococcus agalactiae is a primary pathogen associated with subclinical mastitis in dairy herds, requiring accurate identification and characterization for effective management due to its highly contagious nature. This study evaluated the use of Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) for grouping S. agalactiae isolates from subclinical mastitis in cattle and buffaloes associating the antimicrobial susceptibility profiles of the isolates with the grouping results. A total of 198 milk samples were collected from three farms (Farm A: 67 cow isolates, Farm B: 101 cow isolates, Farm C: 30 buffalo isolates). Antimicrobial susceptibility testing using the Minimum Inhibitory Concentration (MIC) method was performed for 10 antimicrobials. High sensitivity (>90 %) was observed for ceftiofur, penicillin, oxacillin, amoxicillin, cefquinome, gentamicin, and cefoxitin, while low sensitivity (<15 %) was detected for enrofloxacin and cephalexin. Clustering was conducted using Random Amplified Polymorphic DNA (RAPD) and grouped by MALDI-TOF MS. RAPD identified 33 clusters at an 80 % similarity breakpoint, while MALDI-TOF MS identified 8 distinct groups. MALDI-TOF MS successfully grouped all isolates, whereas RAPD clustered only 100. Both methods grouped isolates from the same herd with similar susceptibility profiles. These findings highlight the use of MALDI-TOF MS for rapid grouping pathogens but emphasize differences when compared to RAPD.

01 Apr 2025·The Veterinary Journal

Drug resistance detection of canine origin Escherichia coli in China and inhibition by genipin

Article

Author: Dou, Lei-Na ; Zhuang, Shen ; Zhang, Wei-Min ; Yang, Shuo ; Ma, Wu-Ren ; Liu, Ying-Qiu ; Wang, Hai-Xin ; Niu, Nan ; Fan, Yun-Peng

Zoonotic transmission from pets to their owners is a major health problem. It is important to determine and control the drug resistance in pets to mitigate risks of human transmission. In this work, the current prevalence of multi-drug resistance (MDR) and resistance gene in Escherichia coli (E. coli) derived from dogs in nine cities across various regions of China initially evaluated using microfluidic dilution methods and polymerase chain reaction (PCR) technology. To control antibiotic resistance, genipin as natural product was used to combat MDR E. coli. Finally, the synergistic effect of genipin and norfloxacin on MDR E. coli was studied using time-kill curves to retard the resistance spread. A total of 126 E. coli strains were isolated from 154 collected fecal samples of dogs. Minimum inhibitory concentrations (MIC) results revealed that the highest detection rate of MDR E. coli appeared in Zhengzhou at 90.9 %, and the lowest in Shenyang at 10.0 %. The results of drug resistance gene testing indicated that the bla_TEM gene had the highest detection rate (99.2 %), then tetA and bla_CTX-M-1, whose detection rates all exceed 50 %. Furthermore, the MIC of genipin against MDR E. coli was found to be 4096 μg/mL, and genipin at ½ MIC demonstrated significant inhibition on MDR E. coli within 6 h. Finally, the combination of ¼ MIC genipin with ½ MIC norfloxacin showed partial synergistic inhibitory effect on MDR E. coli. Our findings suggest that although antibiotic resistance in canine origin E. coli varies across different regions of China, it remains concerning, and genipin shows potential as a treatment option for MDR E. coli infections.

01 Mar 2025·Microbiological Research

Emergence of two novel tmexCD-toprJ subtypes mediating tigecycline resistance in the megaplasmids from Pseudomonas putida

Article

Author: Wang, Chengzhen ; Liu, Jian-Hua ; Gao, Xun ; Lv, Luchao ; Zhang, Xiaoyu ; Yue, Chao ; Lu, Litao

The widespread antimicrobial resistance (AMR) problem poses a serious health threat, leaving few drug choices, including tigecycline, to treat multidrug resistance pathogens. However, a plasmid-borne tigecycline resistance gene cluster, tmexCD1-toprJ1, emerged and conferred tigecycline resistance. In this study, we identified two novel subtypes, tmexCD2.3-toprJ2.3 and tmexCD2.4-toprJ1b, obtained from three chicken-origin Pseudomonas putida isolates. Two types of megaplasmids were found as the vital vehicle of these tmexCD-toprJ variants. Phylogenetic and genomic analysis indicated the two variants were mainly distributed in Pseudomonas and acted as an evolved intermediated state precursor of tmexCD2-toprJ2. Further gene cloning assay revealed both the expression of tmexCD2.3-toprJ2.3 and tmexCD2.4-toprJ1b could confer multiple antimicrobial resistance, mediating 8- to 16-fold increase of tigecycline MIC. Importantly, two key nucleotide differences in promoter region influence the promoter activity between P_tmexC2.3 and P_tmexC2.4, while the downregulation effect of TNfxB on the transcriptional expression level of tmexCD2.3-toprJ2.3 and tmexCD2.4-toprJ1b were observed. The emergency of two novel tmexCD-toprJ variants necessitates preventive measures to curb their spread and highlights concerns about more emerging tmexCD-toprJ variants.

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Indication	Highest Phase	Country/Location	Organization	Date
Bacterial Infections	Preclinical	Germany	Hoechst AG	01 Jan 1991

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