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1. Genomic and in vitro pharmacodynamic analysis of rifampicin resistance in multidrug‐resistant canine Staphylococcus pseudintermedius isolates

   https://doi.org/10.1111/vde.12959  

   Hicks, Karly; Tan, Yongjun; Cao, Wenqi; Hathcock, Terri; Boothe, Dawn; Kennis, Robert; Zhang, Dapeng; Wang, Xu; White, Amelia (April 2021, Veterinary Dermatology)

   BackgroundAntimicrobial resistance is a growing concern in canineStaphylococcus pseudintermediusdermatitis. Treatment with rifampicin (RFP) is considered only in meticillin‐resistant and multidrug‐resistantS. pseudintermedius(MDR‐MRSP). Hypothesis/ObjectivesTo determine an optimal RFP dosing for MDR‐MRSP treatment without induction of RFP resistance and identify causal mutations for antimicrobial resistance. Methods and materialsTime–kill assays were performed in a control isolate and three MDR‐MRSP isolates at six clinically relevant concentrations [32 to 1,024 × MIC (the minimum inhibitory concentration)]. Whole‐genome resequencing and bioinformatic analysis were performed in the resistant strains developed in this assay. ResultsThe genomic analysis identified nine antimicrobial resistance genes (ARGs) in MDR‐MRSP isolates, which are responsible for resistance to seven classes of antibiotics. RFP activity against all four isolates was consistent with a time‐dependent and bacteriostatic response. RFP resistance was observed in six of the 28 time–kill assays, including concentrations 64 × MIC in MDR‐MRSP1 isolates at 24 h, 32 × MIC in MDR‐MRSP2 at 48 h, 32 × MIC in MDR‐MRSP3 at 48 h and 256 × MIC in MDR‐MRSP3 at 24 h. Genome‐wide mutation analyses in these RFP‐resistant strains discovered the causal mutations in the coding region of therpoBgene. Conclusions and clinical relevanceA study has shown that 6 mg/kg per os results in plasma concentrations of 600–1,000 × MIC ofS. pseudintermedius. Based on our data, this dose should achieve the minimum MIC (×512) to prevent RFP resistance development; therefore, we recommend a minimum daily dose of 6 mg/kg for MDR‐MRSP pyoderma treatment when limited antibiotic options are available. 

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2. The complete genome sequences of Thioglobus autotrophicus strains EF2 and EF3, isolated from an oxycline in Effingham Inlet, British Columbia

   https://doi.org/10.1128/mra.01118-23  

   Morris, Robert M; Mino, Sayaka (March 2024, Microbiology Resource Announcements)

   Thrash, J Cameron (Ed.)

   ABSTRACT Here we provide the complete genome sequences of two chemoautotrophic isolates from theThioglobaceaefamily of marine gamma-proteobacteria. The genomes were obtained from pure cultures that were initially isolated from Effingham Inlet in 2013 and revived from freezer stocks for whole genome sequencing in 2023. 

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3. Antibiotic resistance trends among Vibrio vulnificus and Vibrio parahaemolyticus isolated from the Chesapeake Bay, Maryland: a longitudinal study

   https://doi.org/10.1128/aem.00539-24  

   Morgado, Michele E; Brumfield, Kyle D; Chattopadhyay, Suhana; Malayil, Leena; Alawode, Taiwo; Amokeodo, Ibiyinka; He, Xin; Huq, Anwar; Colwell, Rita R; Sapkota, Amy R (June 2024, Applied and Environmental Microbiology)

   Elkins, Christopher A (Ed.)

   ABSTRACT Antibiotics are often used to treat severeVibrioinfections, with third-generation cephalosporins and tetracyclines combined or fluoroquinolones alone being recommended by the US Centers for Disease Control and Prevention. Increases in antibiotic resistance of both environmental and clinical vibrios are of concern; however, limited longitudinal data have been generated among environmental isolates to inform how resistance patterns may be changing over time. Hence, we evaluated long-term trends in antibiotic resistance of vibrios isolated from Chesapeake Bay waters (Maryland) across two 3-year sampling periods (2009–2012 and 2019–2022).Vibrio parahaemolyticus(n= 134) andVibrio vulnificus(n= 94) toxR-confirmed isolates were randomly selected from both sampling periods and tested for antimicrobial susceptibility against eight antibiotics using the Kirby-Bauer disk diffusion method. A high percentage (94%–96%) ofV. parahaemolyticusisolates from both sampling periods were resistant to ampicillin and only 2%–6% of these isolates expressed intermediate resistance or resistance to third-generation cephalosporins, amikacin, tetracycline, and trimethoprim-sulfamethoxazole. Even lower percentages of resistantV. vulnificusisolates were observed and those were mostly recovered from 2009 to 2012, however, the presence of multiple virulence factors was observed. The frequency of multi-drug resistance was relatively low (6%–8%) but included resistance against antibiotics used to treat severe vibriosis in adults and children. All isolates were susceptible to ciprofloxacin, a fluoroquinolone, indicating its sustained efficacy as a first-line agent in the treatment of severe vibriosis. Overall, our data indicate that antibiotic resistance patterns amongV. parahaemolyticusandV. vulnificusrecovered from the lower Chesapeake Bay have remained relatively stable since 2009.IMPORTANCEVibriospp. have historically been susceptible to most clinically relevant antibiotics; however, resistance and intermediate-resistance have been increasingly recorded in both environmental and clinical isolates. Our data showed that while the percentage of multi-drug resistance and resistance to antibiotics was relatively low and stable across time, someVibrioisolates displayed resistance and intermediate resistance to antibiotics typically used to treat severe vibriosis (e.g., third-generation cephalosporins, tetracyclines, sulfamethoxazole-trimethoprim, and aminoglycosides). Also, given the high case fatality rates observed withVibrio vulnificusinfections, the presence of multiple virulence factors in the tested isolates is concerning. Nevertheless, the continued susceptibility of all tested isolates against ciprofloxacin, a fluoroquinolone, is indicative of its use as an effective first-line treatment of severeVibriospp. infections stemming from exposure to Chesapeake Bay waters or contaminated seafood ingestion. 

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4. Genomic characterization of Pseudomonas syringae pv. syringae from Callery pear and the efficiency of associated phages in disease protection

   https://doi.org/10.1128/spectrum.02833-23  

   Holtappels, D; Abelson, S A; Nouth, S C; Rickus, G_E J; Amare, S Z; Giller, J P; Jian, D Z; Koskella, B (March 2024, Microbiology Spectrum)

   Zeng, Quan (Ed.)

   ABSTRACT ThePseudomonas syringaespecies complex is a heterogeneous group of plant pathogenic bacteria associated with a wide distribution of plant species. Advances in genomics are revealing the complex evolutionary history of this species complex and the wide array of genetic adaptations underpinning their diverse lifestyles. Here, we genomically characterize twoP. syringaeisolates collected from diseased Callery pears (Pyrus calleryana) in Berkeley, California in 2019 and 2022. We also isolated a lytic bacteriophage, which we characterized and evaluated for biocontrol efficiency. Using a multilocus sequence analysis and core genome alignment, we classified theP. syringaeisolates as members of phylogroup 2, related to other strains previously isolated fromPyrusandPrunus. An analysis of effector proteins demonstrated an evolutionary conservation of effectoromes across isolates classified in PG2 and yet uncovered unique effector profiles for each, including the two newly identified isolates. Whole-genome sequencing of the associated phage uncovered a novel phage genus related toPseudomonas syringaepv.actinidiaephage PHB09 and theFlaumdravirusgenus. Finally, usingin plantainfection assays, we demonstrate that the phage was equally useful in symptom mitigation of immature pear fruit regardless of the Pss strain tested. Overall, this study demonstrates the diversity ofP. syringaeand their viruses associated with ornamental pear trees, posing spill-over risks to commercial pear trees and the possibility of using phages as biocontrol agents to reduce the impact of disease.IMPORTANCEGlobal change exacerbates the spread and impact of pathogens, especially in agricultural settings. There is a clear need to better monitor the spread and diversity of plant pathogens, including in potential spillover hosts, and for the development of novel and sustainable control strategies. In this study, we characterize the first described strains ofPseudomonas syringaepv.syringaeisolated from Callery pear in Berkeley, California from diseased tissues in an urban environment. We show that these strains have divergent virulence profiles from previously described strains and that they can cause disease in commercial pears. Additionally, we describe a novel bacteriophage that is associated with these strains and explore its potential to act as a biocontrol agent. Together, the data presented here demonstrate that ornamental pear trees harbor novelP. syringaepv.syringaeisolates that potentially pose a risk to local fruit production, or vice versa—but also provide us with novel associated phages, effective in disease mitigation. 

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5. Molecular epidemiology and pathogenomics of extended-spectrum beta-lactamase producing- Escherichia coli and - Klebsiella pneumoniae isolates from bulk tank milk in Tennessee, USA

   https://doi.org/10.3389/fmicb.2023.1283165  

   Gelalcha, Benti D; Mohammed, Ruwaa I; Gelgie, Aga E; Kerro_Dego, Oudessa (November 2023, Frontiers in Microbiology)

   IntroductionThe rise in extended-spectrum beta-lactamase (ESBL)-producingEnterobacteriaceaein dairy cattle farms poses a risk to human health as they can spread to humans through the food chain, including raw milk. This study was designed to determine the status, antimicrobial resistance, and pathogenic potential of ESBL-producing -E. coliand -Klebsiellaspp. isolates from bulk tank milk (BTM). MethodsThirty-three BTM samples were collected from 17 dairy farms and screened for ESBL-E. coliand -Klebsiellaspp. on CHROMagar ESBL plates. All isolates were confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and subjected to antimicrobial susceptibility testing and whole genome sequencing (WGS). ResultsTen presumptive ESBL-producing bacteria, eightE. coli, and twoK. pneumoniaewere isolated. The prevalence of ESBL-E. coliand -K. pneumoniaein BTM was 21.2% and 6.1%, respectively. ESBL-E. coliwere detected in 41.2% of the study farms. Seven of the ESBL-E. coliisolates were multidrug resistant (MDR). The two ESBL-producingK. pneumoniaeisolates were resistant to ceftriaxone. Seven ESBL-E. colistrains carry thebla_CTX-Mgene, and five of them co-harboredbla_TEM-1. ESBL-E. colico-harboredbla_CTX-Mwith other resistance genes, includingqnrB19,tet(A),aadA1,aph(3’’)-Ib,aph(6)-Id),floR,sul2, and chromosomal mutations (gyrA, gyrB, parC, parE, and pmrB). MostE. coliresistance genes were associated with mobile genetic elements, mainly plasmids. Six sequence types (STs) ofE. coliwere detected. All ESBL-E. coliwere predicted to be pathogenic to humans. Four STs (three ST10 and ST69) were high-risk clones ofE. coli. Up to 40 virulence markers were detected in allE. coliisolates. One of theK. pneumoniaewas ST867; the other was novel strain.K. pneumoniaeisolates carried three types of beta-lactamase genes (bla_CTX-M,bla_TEM-1andbla_SHV). The novelK. pneumoniaeST also carried a novel IncFII(K) plasmid ST. ConclusionDetection of high-risk clones of MDR ESBL-E. coliand ESBL-K. pneumoniaein BTM indicates that raw milk could be a reservoir of potentially zoonotic ESBL-E. coliand -K. pneumoniae. 

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