Screening For Resistance Research Articles

Molecular typing of reduced susceptibility of Acinetobacter calcoaceticus-baumannii complex to Chlorhexidine in Turkey by pulsed-field gel electrophoresis.

Introduction. The global spread of Acinetobacter spp., particularly the Acinetobacter calcoaceticusbaumannii (ACB) complex, has led to its recognition as a significant pathogen by the World Health Organization (WHO). The increasing resistance of the ACB complex to multiple antibiotics presents a challenge for treatment, necessitating accurate antibiotic susceptibility profiling after isolation.Hypothesis or gap statement. There is limited understanding of the antimicrobial resistance and chlorhexidine, a biocide, susceptibility profiles of ACB complex strains, especially in clinical settings in Turkey.Aim. This study aimed to identify ACB complex strains recovered from various clinical specimens at Hacettepe University Hospitals in Ankara, Turkey, in 2019, and to assess identification, their antibiotic and chlorhexidine susceptibility profiles, and genomic relatedness.Methodology. Eighty-two ACB complex strains were identified using MALDI-TOF MS. Susceptibility testing to 12 antibiotics was conducted using the disc diffusion method, and colistin, chlorhexidine susceptibility was assessed using the broth microdilution technique, following the latest EUCAST and CLSI guidelines. ACB complex members with reduced chlorhexidine sensitivity were further analyzed by pulsed-field gel electrophoresis (PFGE) for bacterial typing.Results. Among the isolates, 1.2% were multidrug-resistant (MDR), 73.2% were extensively drug-resistant (XDR), and 12.2% were pandrug-resistant (PDR). Carbapenem resistance was found in 86.7% of MDR, PDR, and XDR strains. Colistin resistance was observed in 15.8% of isolates, and 18.2% exhibited decreased susceptibility to chlorhexidine. PFGE revealed seven different clones among strains with reduced chlorhexidine sensitivity, indicating vertical transmission within the hospital.Conclusion. This study highlights the reduced susceptibility to chlorhexidine in ACB complex members and provides epidemiological insights into their spread. The findings underscore the importance of screening for antimicrobial resistance and biocide susceptibility profiles to effectively manage healthcare-associated infections.

Rapid screening of creep resistance in additive manufactured Ti-6Al-4V alloy

In this study, rapid screening of creep resistance for additively manufactured (AM) materials is enabled using an accelerated creep test (ACT) called the dynamic negative stepped test (DNST). Additively manufactured components are on the rise in aerospace systems, particularly turbine engine parts. It can be challenging to identify the AM process parameters that correspond to high-temperature strength and creep resistance. Tests are needed that rapidly screen and qualify creep resistance. In DNST, the temperature is held constant, and the specimen is taken through a series of load holds from high to low. The DNST contains two distinct characteristics: (1) the hold duration for each step is “dynamically” controlled to ensure the minimum-creep-strain-rate is achieved at every step, and (2) the “descending” profile maximizes damage accumulation while remaining on the edge of instability. The DNST assesses a material's ability to resist creep damage accumulated across multiple deformation mechanisms. In this study, electron beam melted Ti–6Al–4V, built at six (6) directions: 0°, 30°, 45°, 60°, 90°, and V, is tested using a four-step DNST at 650 °C. Analysis of the minimum-creep-strain-rate (at each step), creep ductility, and rupture were performed. A strain energy density (SED) benchmark metric was established to rapidly screen and classify creep resistance using an “Ashby-like” plot that balanced energy dissipated and energy dissipation rate. The creep resistance was found to be ranked from V, 60, 45, 30, 90, and 0 in descending order. Microstructural and crystallographic characterization revealed grain refinement progressing from the strain-free to the high-strain regions, indicative of dynamic recrystallization (DRX). Further observations revealed that the material's energy capacity increased with the alignment of the grains' long axis and tensile direction.

A Quantitative Index for Evaluating Maize Leaf Wilting and Its Sustainable Application in Drought Resistance Screening

A Quantitative Index for Evaluating Maize Leaf Wilting and Its Sustainable Application in Drought Resistance Screening

Azole resistance screening in Aspergillus fumigatus sensu stricto using the azole-containing agar method (EUCAST E.Def 10.2): conidial suspension filtration and inoculum adjustment before inoculum preparation may not be needed.

Azole resistance screening in Aspergillus fumigatus sensu stricto can be routinely carried out by using azole-containing plates (E.Def 10.2 procedure); however, conidial suspension filtering and inoculum adjustment before inoculation of plates are time-consuming. We, here, showed that unfiltered and unadjusted conidial suspensions do not negatively influence the performance of the E.Def 10.2 method when screening for azole resistance in A. fumigatus sensu stricto.

Biological prediction model of insulin resistance in obese adolescents based on LASSO-Logistic regression

To identify risk factors affecting the development of insulin resistance in obese adolescents, and to build a nomograph model for predicting the risk of insulin resistance and achieve early screening of insulin resistance. A total of 404 obese adolescents aged 10 to 17 years were randomly recruited through a weight loss camp for the detection and diagnosis of lipids and insulin resistance between 2019 and 2021, and key lipid indicators affecting the development of insulin resistance were screened by Lasso regression, nomogram model was constructed, and internal validation of the models was performed by Bootstrap method, and the area under the working characteristic curve(ROC-AUC) and clinical decision curve were used to assess the calibration degree and stability of the column line graph. The AUC was 0.825(95% CI 0.782-0.868), the internal validation result C-Index was 0.804, the mean absolute error of the column line graph model to predict the risk of insulin resistance was 0.015 and the Brier score was 0.163. The Hosmer-Lemeshow goodness-of-fit test showed that model is ideal and acceptable(χ~2=5.59, P=0.70). The nomogram model of triglyceride, low-density lipoprotein cholesterol and total cholesterol/high-density lipoprotein cholesterol based on Lasso-logistic regression can effectively predict the risk of insulin resistance in obese children and adolescents.

Screening of Drought-Resistant Potato Germplasm Resources Based on Intelligent Algorithm

As a staple food resource, potato is of great significance for improving grain reserves and ensuring national food security. In order to improve potato yield and promote the process of potato becoming a staple food, a K-means algorithm optimized by particle swarm algorithm was proposed to realize the screening of dry potato germplasm resources. First, the research continues the research on particle swarm optimization, and innovatively applies K-means algorithm to optimization. The research utilizes the advantages of particle swarm optimization, such as fast convergence speed, strong search ability, and simple operation, to enable particle swarm optimization to take on the role of optimizing the initial clustering center, thereby improving the accuracy and efficiency of clustering analysis. On this basis, a PSO-K-means drought resistant potato germplasm resource screening model was constructed. This model consists of a data collection and preprocessing module, an impact indicator determination module, and a comprehensive evaluation module. Finally, the application effect of the model was verified. The results show that the AUC value of the model is up to 0.840, and the screening accuracy is as high as 94.5%, which is 13.5% higher than that of the K-means model. The research method has been validated to improve the limitations of K-means mode, such as high screening error, weak stability, and falling into local optimal solutions. It optimizes the screening effect of drought resistant potato germplasm resources, which is conducive to exploring the potential of potato resources. In addition, research has also provided broader ideas for the optimization and application of particle swarm optimization algorithms.

IFNγ mediates the resistance of tumor cells to distinct NK cell subsets

BackgroundImmune checkpoint blockade targeting the adaptive immune system has revolutionized the treatment of cancer. Despite impressive clinical benefits observed, patient subgroups remain non-responsive underscoring the necessity for combinational therapies harnessing...

Screening for Plasmid Mediated Antibiotic Resistance among Multi Drug Resistant Bacteria Isolated from Patients with Lower Respiratory Tract Infections Attending some Hospitals in Kano Metropolis

Study’s Novelty/Excerpt This study investigates the prevalence and plasmid-mediated resistance of multidrug-resistant (MDR) bacteria isolated from patients with lower respiratory tract infections (LRTIs), revealing significant findings pertinent to public health. The research demonstrates a high prevalence of MDR isolates (20%) and highlights the substantial role of plasmids, with 67.6% of MDR isolates containing plasmids, and 80% losing drug resistance post-plasmid curing. These results underscore the necessity for treatment options based on antibiotic susceptibility testing and continuous plasmid profiling to combat plasmid-mediated resistance effectively, marking a critical advancement in the management of LRTIs. Full Abstract Lower Respiratory tract infections (LRTIs) are among the serious infections in humans that have been worsening by the emergence of drug resistant bacteria and aggravated by plasmids transfer. The study aimed to screen Multidrug-resistant (MDR) bacteria isolated from patients with LRTIs for the presence of Plasmid. Sputum samples (400) were collected from patients with LRTIs and processed using standard microbiological procedures to isolate lower respiratory tract bacteria. The identified isolates were subjected to antibacterial susceptibility testing using the disc diffusion method. Multidrug resistant isolates were further subjected to plasmid curing using Acridine orange and the cured isolates were tested for loss of drug resistance. Among the 400 samples, 185 (46.2 %) harbored significant bacterial growth, with 83 (44.9%) Gram-positive and 102 (55.1%) Gram-negative bacteria. Bacteria isolated include Escherichia coli (29), Klebsiella pneumonia (46), Moraxella catarrhalis (13), Pseudomonas aerugonosa (14), Staphylococcus aureus(24) and Streptococcus pneumoniae (59). A higher infection rate was recorded in males (46.99%) and patients aged 41-50 (52.59%). The highest resistance was exhibited by Klebsiella pneumoniae (93%) against cefuroxime, followed by Streptococcus pneumoniae against oxacillin (75%). Of the 185 isolates, 37 (20%) were MDR, out of which 25 (67.6%) isolates had Plasmids. Following curing, 20 (80%) of the isolates were cured, and only 5 (20%) retained their plasmids. Most importantly, Escherichia coli and Klebsiella pneumonia were found to be sensitive to levofloxacin, gentamicin, and imipenem but retained their resistance to Cefuroxime, while Streptococcus pneumoniae was found to be sensitive to levofloxacin only with the highest resistance to oxacillin, doxycycline, and erythromycin. The study establishes a high prevalence of MDR isolates among LRTIs, with some exhibiting plasmid-mediated resistance, and therefore recommends treatment options to be solely based on antibiotic susceptibility testing and continuous plasmid profiling to detect plasmid-mediated resistance to enable appropriate drug administration.

Thermal time and time of sowing impacts disease development of Brassica napus inoculated with diverse Sclerotinia sclerotiorum isolates

AbstractSclerotinia stem rot (SSR), caused by the necrotrophic fungal pathogen Sclerotinia sclerotiorum, is a severe disease of broadleaf crops including canola/rapeseed (Brassica napus), leading to significant yield loss in conducive years. Replication of field conditions is challenging in variety disease resistance screening with testing required under a wide range of environmental conditions and at different plant growth stages. We investigated the role of thermal time in disease progression using three sowing times in the growing season, with six commonly grown Australian varieties of canola inoculated with four diverse West Australian isolates of S. sclerotiorum at 30% flowering. Area Under the Disease Progression Stairs (AUDPS), sclerotia production and weight, and seed production were measured. Time of sowing was found to be a crucial factor in explaining differences in AUDPS, stem width, seed production and sclerotia number according to the analysis of variance (P < 0.05), influencing isolate aggressiveness and disease progression. Linear mixed-effect models, regression decision tree models and principal components analysis were also conducted to determine the importance of a range of variables being included in variety screening for resistance. For all these analyses, both thermal time from sowing to 30% flowering when plants were inoculated, as well as thermal time over the 28-day inoculation period, in both canola and S. sclerotiorum, were important in explaining the variation. The study concludes by recommending that thermal time should be included in future SSR prediction risk models.

Microneedle assisted delivery of the cloned components of cucurbit leaf crumple virus in yellow squash (Cucurbita pepo)

Cucurbit leaf crumple virus (CuLCrV) is among the prominent viruses infecting cucurbits in the USA. Attainable procedures of virus inoculation to crops are prerequisite for screening of resistance against the virus. Because mechanical (non-vector-mediated) infection by cucurbit leaf crumple virus (CuLCrV) is inefficient in economically important crops, screening for CuLCrV resistance is currently laborious and time-consuming using transmission by viruliferous whiteflies. We constructed an infectious partial tandem repeat construct of an isolate of CuLCrV from Georgia, USA, in the plant expression binary vector pCambia2300 and transformed it into Agrobacterium tumifaciens strain EHA105. Agroinfiltration of this construct into the abaxial surface of the leaves of common bean (Phaseolus vulgaris L.) produced a systemic infection characteristic of CuLCrV, although this approach was not successful for yellow squash. However, we report a very efficient and reproducible inoculation procedure established in squash when the leaves were injured with a microneedle and rubbed it with cell suspension harbouring the infectious viral construct.

Screening for antifolate and artemisinin resistance in Plasmodium falciparum dried-blood spots from three hospitals of Eritrea

Background Antimalarial drug resistance is a major challenge hampering malaria control and elimination. About three-quarters of Eritrea’s population resides in the malaria-endemic western lowlands of the country. Plasmodium falciparum, the leading causative parasite species, has developed resistance to basically all antimalarials. Continued surveillance of drug resistance using genetic markers provides important molecular data for treatment policies which complements clinical studies, and strengthens control efforts. This study sought to genotype point mutations associated with P. falciparum resistance to sulfadoxine-pyrimethamine and artemisinin, in dried-blood spots from three hospitals in the western lowlands of Eritrea. Methods Dried-blood spot samples were collected from patients visiting Adi Quala, Keren and Gash Barka Hospitals, between July and October, 2014. The patients were followed up after treatment with first line artesunate-amodiaquine, and dried-blood spots were collected on day three after treatment. Nested polymerase chain reaction and Sanger sequencing techniques were employed to genotype point mutations in the Pfdhfr (PF3D7_0417200), Pfdhps (PF3D7_0810800) and PfK13 (PF3D7_1343700) partial gene regions. Results Sequence data analyses of PCR-positive isolates found wild-type artemisinin haplotypes associated with resistance (Y493Y, R539R, I543I) in three isolates, whereas four mutant antifolate haplotypes associated with resistance were observed in six isolates. These included the triple-mutant Pfdhfr (S108N, C59R, N51I) haplotype, the double-mutant Pfdhfr (N51I, S108N) haplotype, the single-mutant Pfdhfr (K540E) haplotype, and the mixed-mutant Pfdhfr-Pfdhps (S108N, N51I + K540E) haplotype. Other findings observed were, a rare non-synonymous Pfdhfr V45A mutation in four isolates, and a synonymous Pfdhps R449R in one isolate. Conclusions The mutant antifolate haplotypes observed indicate a likely existence of full SP resistance. Further studies can be carried out to estimate the prevalence of SP resistance. The wild-type artemisinin haplotypes observed suggest artemisinin is still an effective treatment. Continuous monitoring of point mutations associated with delayed parasite clearance in ART clinical studies is recommended.

Azole Resistance in Veterinary Clinical Aspergillus fumigatus Isolates in the Netherlands

Aspergillus fumigatus is a saprophytic fungal pathogen that causes opportunistic infections in animals and humans. Azole resistance has been reported globally in human A. fumigatus isolates, but the prevalence of resistance in isolates from animals is largely unknown. A retrospective resistance surveillance study was performed using a collection of clinical A. fumigatus isolates from various animal species collected between 2015 and 2020. Agar-based azole resistance screening of all isolates was followed by in vitro antifungal susceptibility testing and cyp51A gene sequencing of the azole-resistant isolates. Over the 5 year period 16 (11.3%) of 142 A. fumigatus culture-positive animals harbored an azole-resistant isolate. Resistant isolates were found in birds (15%; 2/13), cats (21%; 6/28), dogs (8%; 6/75) and free-ranging harbor porpoise (33%; 2/6). Azole-resistance was cyp51A mediated in all isolates: 81.3% (T-67G/)TR34/L98H, 12.5% TR46/Y121F/T289A. In one azole-resistant A. fumigatus isolate a combination of C(-70)T/F46Y/C(intron7)T/C(intron66)T/M172V/E427K single-nucleotide polymorphisms in the cyp51A gene was found. Of the animals with an azole-resistant isolate and known azole exposure status 71.4% (10/14) were azole naive. Azole resistance in A. fumigatus isolates from animals in the Netherlands is present and predominantly cyp51A TR-mediated, supporting an environmental route of resistance selection. Our data supports the need to include veterinary isolates in resistance surveillance programs. Veterinarians should consider azole resistance as a reason for therapy failure when treating aspergillosis and consider resistance testing of relevant isolates.

Displacement of Hospital-Acquired, Methicillin-Resistant Staphylococcus aureus Clones by Heterogeneous Community Strains in Kenya over a 13-Year Period.

We determined antibiotic susceptibility and employed Oxford Nanopore whole-genome sequencing to explore strain diversity, resistance, and virulence gene carriage among methicillin-resistant Staphylococcus aureus (MRSA) strains from different infection sites and timepoints in a tertiary Kenyan hospital. Ninety-six nonduplicate clinical isolates recovered between 2010 and 2023, identified and tested for antibiotic susceptibility on the VITEK ID/AST platform, were sequenced. Molecular typing, antibiotic resistance, and virulence determinant screening were performed using the relevant bioinformatics tools. The strains, alongside those from previous studies, were stratified into two periods covering 2010-2017 and 2018-2023 and comparisons were made. Mirroring phenotypic profiles, aac(6')-aph(2″) [aminoglycosides]; gyrA (S84L) and grlA (S80Y) [fluoroquinolones]; dfrG [anti-folates]; and tet(K) [tetracycline] resistance determinants dominated the collection. While the proportion of ST239/241-t037-SCCmec III among MRSA reduced from 37.7% to 0% over the investigated period, ST4803-t1476-SCCmec IV and ST152-t355-SCCmec IV were pre-eminent. The prevalence of Panton-Valentine leucocidin (PVL) and arginine catabolic mobile element (ACME) genes was 38% (33/87) and 6.8% (6/87), respectively. We observed the displacement of HA-MRSA ST239/241-t037-SCCmec III with the emergence of ST152-t355-SCCmec IV and a greater clonal heterogeneity. The occurrence of PVL+/ACME+ CA-MRSA in recent years warrants further investigations into their role in the CA-MRSA virulence landscape, in a setting of high PVL prevalence.

MSK-ACCESS powered with SOPHiA DDM: Performance analysis of a decentralized MSK-ACCESS solution.

e15063 Background: Blood plasma cell-free DNA (cfDNA) testing of cancer patients allows for genomic profiling when tissue sampling is too invasive or of insufficient quality. This approach has demonstrated clinical utility to identify treatment targets, screen for resistance and monitor disease burden. MSK-ACCESS (Memorial Sloan Kettering – Analysis of Circulating cfDNA to Evaluate Somatic Status) is a validated, high sensitivity cfDNA assay employed at MSK to identify biologically actionable tumor alterations in clinical care. SOPHiA GENETICS is developing a decentralized solution of this assay that incorporates major features of MSK-ACCESS such as molecular barcoding, duplex-based variant calling, the inclusion of tumor-informed prior knowledge and identification of somatic alterations through the removal of both germline and clonal hematopoiesis (CH) variants. Here, we present the results of a performance study obtained during product development. Methods: A cohort of 24 plasma cfDNA and matched white blood cell (WBC) DNA samples were collected from patients with solid tumors and processed at MSK according to the validated standard operating procedures and bioinformatic analyses. In parallel, for the same set of materials libraries were constructed at SOPHiA GENETICS using the proprietary CUMIN molecular identifiers, captured with probes targeting the updated MSK-ACCESS v2 set of 146 genes and sequenced on an Illumina NovaSeq 6000. Variants were called using a duplex-aware approach by SOPHiA GENETICS and germline and CH variants were identified and filtered by paired cfDNA – WBC DNA analysis. Accuracy of the decentralized “MSK-ACCESS powered with SOPHiA DDM” solution was assessed through concordance to the orthogonal results obtained by MSK. Results: A total of 104 somatic SNV/Indels were detected with variant allele fractions (VAF) ranging from 0.1% to 97.0% with a median of 5 variants per sample. The positive percent agreement (PPA) for all variants above the limit of detection of the original MSK-ACCESS assay was 98.9% with 99.98% specificity. For the subset of prior-knowledge variants PPA was 95.6% down to 0.1% VAF. A total of additional 59 variants with ≤ 15% VAF were found both in cfDNA and WBC and assigned as CH-derived. Importantly, these occurred not only in CH-associated genes but also in common oncogenes such as BRCA1/2 and ERBB2, highlighting the utility of matched WBC analysis to correctly identify somatic variants. Conclusions: The results demonstrate that the decentralized “MSK-ACCESS powered with SOPHiA DDM” solution exhibits a high degree of agreement to the results achieved by MSK. Further studies will be conducted to expand the analytical performance evaluation.

QAPI 143 - Modifying Multi-Drug Resistant Organism Admission Screening to Improve Policy Compliance

QAPI 143 - Modifying Multi-Drug Resistant Organism Admission Screening to Improve Policy Compliance

Comparison of two different experimental environments for resistance screenings for the leafhopper-transmitted wheat dwarf virus in wheat

Wheat dwarf virus (WDV) causes high yield losses in wheat and other cereals and is therefore an important pathogen transmitted by the leafhopper Psammotettix alienus. Climate change will increase infections by insect-transmitted viruses due to the increasing spread of vectors. In the context of integrated pest management, the cultivation of WDV-resistant/tolerant varieties is an effective way of controlling WDV. Evaluation of tolerant/resistant genotypes is based on inoculation with viruliferous leafhoppers and subsequent phenotyping in gauze houses under semi-field conditions. For successful screening, it is important to ensure the uniform and reproducible inoculation of plants. Abiotic conditions, particularly temperature, have a critical influence on inoculation success, and thus, variations in infection rates were observed within and between previous replicates in the field. Furthermore, the leafhopper population reared in the greenhouse has to be reestablished after each infection, which delays the screening process. We addressed these issues by developing an improved inoculation assay in which plants are inoculated in small infection hoods in the greenhouse before being planted out in gauze houses. This procedure allows optimal environmental conditions for WDV infection of test plants and allows the plants with WDV infection to develop under natural environmental conditions for symptom scoring. In addition, the viruliferous leafhoppers were recollected from the test plants after infection, allowing a sustainable use of the insects. The method thus enables more reliable phenotyping by increasing infection success and testing a greater number of genotypes in a shorter time.

Macrolide and fluoroquinolone resistance associated mutations in Mycoplasma genitalium in men who have sex with men attending STI clinic: A pilot study from India.

Background Increasing rates of macrolide and fluroquinolone resistance in Mycoplasma genitalium (MG) are being reported worldwide with resultant treatment failure. Aim We aimed to determine the level of antibiotic resistance of MG in men who have sex with men (MSM) attending a sexually transmitted infections (STIs) clinic in New Delhi, India. Methods Real-time polymerase chain reaction (PCR) assays targeting MgPa and pdhD genes were performed to detect MG rectal, urogenital or oropharyngeal infections in 180 MSM between January 2022 and June 2023. Macrolide resistance-associated mutations (MRM) and quinolone resistance-associated mutations (QRM) were detected by specific amplification of domain V of 23SrRNA gene and appropriate regions of parC and gyrA genes respectively followed by sequencing. PCR-based screening for Chlamydia trachomatis (CT) infection was also performed. Results A total of 13 (7.2%) MSM were positive for MG infection. The most common site of infection was anorectum (8/13; 61.5%) followed by the urethra (5/13; 38.5%). None of the patients had infection at both the sites, and no oropharyngeal MG infection was detected. CT infection was detected in 37 (20.6%) MSM. Of the 13 MG-infected MSM, 6 (46.2%) were co-infected with CT. MRM and QRM were found in five (46.2%) and two (15.4%) strains, respectively. Both Quinolone resistance mutation (QRM)-harbouring strains also harboured MRM. All the five MG isolates carried the MRM A2071G. Both the QRM isolates co-harboured the parC and gyrA single-nucleotide polymorphisms. There was no correlation between the presence of antibiotic resistance and co-infection with CT (P = 0.52). Limitation Because all patients in the study were MSM, the high rate of resistance to macrolides and fluoroquinolones could not be extrapolated for non-MSM patients. Conclusion This is a report of an initial survey of antibiotic resistance to MG in a country where its diagnosis and treatment are not routinely available. We found a high prevalence of MG-carrying MRM, QRM and dual-class resistance in MSM in the absence of antibiotic exposure. This study mandates the need for both screening and detection of antimicrobial resistance against MG.

Occurrence and antimicrobial resistance pattern of Escherichia coli and Salmonella species isolated from domestic and peridomestic rodents

E. coli and Salmonella species are bacterial pathogens contributing to opportunistic infections, food poisoning, and urinary tract infections. Recently, research has shown that pathogens have repeatedly been found in rodents. Environmental interaction between human, livestock, and rodents is possibly disseminating these bacteria and their antimicrobial resistance. Therefore, this study aimed to determine the prevalence and antimicrobial resistance patterns of E. coli and Salmonella spp. isolated from domestic and peri-domestic rodents in Iringa municipality, Tanzania. A total of 153 rodents were captured from January to March 2023 and clarified at the species level using identification keys. Rectal swabs were collected. E. coli and Salmonella spp. were isolated by culture methods and identified by biochemical tests and conventional PCR. The antimicrobial susceptibility test was done by disk diffusion method using five antimicrobials, including tetracycline, ciprofloxacin, gentamicin, ceftriaxone, and trimethoprim-sulfamethoxazole. Seven resistant genes were tested by PCR, which were blaCTX-M, blaSHV, sul1, sul2, tetA, acrA, and aac(3)-1. Three rodent species, Rattus rattus (75.2%), Mastomys natalensis (23.5%), and Mus musculus (1.3%), were captured. 17 (11.1%) E. coli were detected, and no Salmonella spp. were isolated. All 17 isolates were susceptible to gentamycin and resistant to sulphamethoxazole/trimethoprim (52.9%). Four E. coli isolates exhibited multidrug resistance (MDR), whereby 75% of these MDR isolates originated from the same area. Six resistant genes were detected: blaCTX-M, sul1, sul2, tetA, acrA, and aac(3)-1, where the acr(A) resistant gene was the most abundant. There were co-occurrences of the resistant genes per isolate, such as sul2, acrA, and aac(3)-1. This study reveals the antimicrobial resistance of E. coli isolated from rodents, providing preliminary data on the prevalence and antimicrobial resistant features of these pathogens in their respective reservoirs in Iringa municipality. The study recommends observing regular antimicrobial resistance screening and improving rodent management and control programs in the studied area.

Toxoplasma replication is inhibited by MMV676477 without development of resistance.

Protozoan parasites cause life-threatening infections in both humans and animals, including agriculturally significant livestock. Available treatments are typically narrow spectrum and are complicated by drug toxicity and the development of resistant parasites. Protozoan tubulin is an attractive target for the development of broad-spectrum antimitotic agents. The Medicines for Malaria Pathogen Box compound MMV676477 was previously shown to inhibit replication of kinetoplastid parasites, such as Leishmania amazonensis and Trypanosoma brucei, and the apicomplexan parasite Plasmodium falciparum by selectively stabilizing protozoan microtubules. In this report, we show that MMV676477 inhibits intracellular growth of the human apicomplexan pathogen Toxoplasma gondii with an EC50 value of ~50 nM. MMV676477 does not stabilize vertebrate microtubules or cause other toxic effects in human fibroblasts. The availability of tools for genetic studies makes Toxoplasma a useful model for studies of the cytoskeleton. We conducted a forward genetics screen for MMV676477 resistance, anticipating that missense mutations would delineate the binding site on protozoan tubulin. Unfortunately, we were unable to use genetics to dissect target interactions because no resistant parasites emerged. This outcome suggests that future drugs based on the MMV676477 scaffold would be less likely to be undermined by the emergence of drug resistance.

Proteome analysis, genetic characterization, and antibiotic resistance patterns of Klebsiella pneumoniae clinical isolates

Klebsiella pneumoniae (K. pneumoniae) is a member of the ESKAPE group and is responsible for severe community and healthcare-associated infections. Certain Klebsiella species have very similar phenotypes, which presents a challenge in identifying K. pneumoniae. Multidrug-resistant K. pneumoniae is also a serious global problem that needs to be addressed. A total of 190 isolates were isolated from urine (n = 69), respiratory (n = 52), wound (n = 48) and blood (n = 21) samples collected from various hospitals in the Al-Qassim, Saudi Arabia, between March 2021 and October 2022. Our study aimed to rapidly and accurately detect K. pneumoniae using the Peptide Mass Fingerprinting (PMF) technique, confirmed by real-time PCR. Additionally, screening for antibiotic susceptibility and resistance was conducted. The primary methods for identifying K. pneumoniae isolates were culture, Gram staining, and the Vitek® 2 ID Compact system. An automated MALDI Biotyper (MBT) instrument was used for proteome identification, which was subsequently confirmed using SYBR green real-time polymerase chain reaction (real-time PCR) and microfluidic electrophoresis assays. Vitek® 2 AST-GN66 cards were utilized to evaluate the antimicrobial sensitivity of K. pneumoniae isolates. According to our results, Vitek® 2 Compact accurately identified 178 out of 190 (93.68%) K. pneumoniae isolates, while the PMF technique correctly detected 188 out of 190 (98.95%) isolates with a score value of 2.00 or higher. Principal component analysis was conducted using MBT Compass software to classify K. pneumoniae isolates based on their structure. Based on the analysis of the single peak intensities generated by MBT, the highest peak values were found at 3444, 5022, 5525, 6847, and 7537 m/z. K. pneumoniae gene testing confirmed the PMF results, with 90.53% detecting entrobactin, 70% detecting 16 S rRNA, and 32.63% detecting ferric iron uptake. The resistance of the K. pneumoniae isolates to antibiotics was as follows: 64.75% for cefazolin, 62.63% for trimethoprim/sulfamethoxazole, 59.45% for ampicillin, 58.42% for cefoxitin, 57.37% for ceftriaxone, 53.68% for cefepime, 52.11% for ampicillin-sulbactam, 50.53% for ceftazidime, 52.11% for ertapenem, and 49.47% for imipenem. Based on the results of the double-disk synergy test, 93 out of 190 (48.95%) K. pneumoniae isolates were extended-spectrum beta-lactamase. In conclusion, PMF is a powerful analytical technique used to identify K. pneumoniae isolates from clinical samples based on their proteomic characteristics. K. pneumoniae isolates have shown increasing resistance to antibiotics from different classes, including carbapenem, which poses a significant threat to human health as these infections may become difficult to treat.