Multidrug-resistant Isolates Research Articles

Polymyxin B adjuvants against polymyxin B- and carbapenem-resistant Gram-negative bacteria.

Aim: Polymyxin B (PMB) is one of the few therapeutic options for treating infections caused by carbapenem-resistant Gram-negative bacteria (CR-GNB). However, the emergence of PMB-resistant CR-GNB strains has prompted the exploration of antibiotic adjuvants as potential therapeutic avenues. Thus, this study evaluates the potential of 3,5-dinitrobenzoic acid derivatives (DNH01, DNH11, DNH13 and DNH20) and isoniazid-N-acylhydrazones (INZ1-7, INZ9 and INZ11) as adjuvants to enhance PMB efficacy against CR-GNB.Materials & methods: MIC, MBC and drug combination assays were conducted using multidrug-resistant clinical isolates of Enterobacterales and Acinetobacter baumannii. In addition, the effects of PMB and PMB+DNH derivatives were assessed through flow cytometry and scanning electron microscopy (SEM).Results: DNH01, DNH11 and DNH20, unlike the INH-acylhydrazones, significantly restored PMB activity (MIC≤2μg/ml) in 80% of the tested isolates. Flow cytometry and SEM assays confirmed that DNH derivatives rescued the activity of PMB, yielding results comparable to those expected for PMB alone but at 256-fold lower concentrations.Conclusion: These findings suggest DNH derivatives hold substantial promise as PMB adjuvants to combat PMB-resistant CR-GNB infections.

Prevalence of qacEΔ1, qacE, oqxA, oqxB, acrA, cepA and zitB genes among multidrug-resistant Klebsiella pneumoniae isolated in a cardiac hospital

Background. Infections caused by multidrug-resistant (MDR) Klebsiella pneumoniae are the leading cause of mortality worldwide. The widespread use of disinfectants and antiseptics has caused the emergence of K. pneumoniae with reduced sensitivity to them, which, in combination with MDR, can pose a significant epidemiological threat. The aim of the study was to assess the prevalence of efflux pump and transporter genes associated with biocide resistance and their association with antibiotic resistance among K. pneumoniae isolated in a cardiac surgical hospital. Materials and methods. K. pneumoniae isolates (n = 50) from the patients and medical equipment were tested by polymerase chain reaction for the presence of genes of 4 types of efflux pumps (qacEΔ1, qacE, oqxA, oqxB, acrA) and 2 transporters involved in the outflow of cations (cepA) and zinc ions (zitB). Spearman's rank correlation test was used to assess the strength of the association between the efflux pumps, beta-lactamase genes and mobile genetic elements. Results. The occurrence of K. pneumoniae containing qacEΔ1, qacE, oqxA, oqxB, acrA, cepA and zitB was high: 54, 62, 100, 84, 100, 72 и 96% respectively. K. pneumoniae with a combination of all the studied pumps was most often detected (32%), and these isolates were MDR in 100% of cases. The qacE, qacEΔ1 genes were closely associated with resistance to cephalosporins, carbapenems, fluoroquinolones, carbapenemase genes, and integrons. The results of the study showed that the genes of various efflux pumps associated with biocide resistance and their combinations were widely represented among the clinical isolates of MDR K. pneumoniae. Conclusion. The high prevalence of efflux pump genes associated with resistance to quaternary ammonium compounds, chlorhexidine and zinc salts and their significant association with antibiotic resistance in nosocomial K. pneumoniae underlines the importance of further studying the mechanisms of cross-resistance to biocides to improve methods of combating MDR nosocomial pathogens.

Raw meat diets are a major risk factor for carriage of third-generation cephalosporin-resistant and multidrug-resistant E. coli by dogs in the UK.

Raw-meat diets (RMD) for dogs, comprising unprocessed or non-heat-treated animal material, are increasingly popular. However, RMDs have been demonstrated to be contaminated with antimicrobial resistant (AMR) bacteria, and there is concern that such diets may pose a zoonotic disease risk. Additionally, dogs fed RMD may shed more AMR- fecal bacteria compared to those fed conventional cooked diets. Data from the UK remain limited; the present study investigated the presence of AMR-Escherichia coli in the feces of RMD and non-RMD (NRMD)-fed dogs in the UK, the E. coli AMR gene complement, and the lifestyle risk factors associated with AMR- E. coli carriage. Fecal samples from UK-owned dogs (N = 193 RMD, N = 239 NRMD) and questionnaires discussing lifestyle factors, were obtained between October 2020-August 2021. Samples underwent culture and antimicrobial susceptibility testing to determine the presence of AMR-E. coli. Whole genome sequencing determined AMR gene carriage. Risk factors for the presence of AMR-E. coli were determined by multivariable modeling. RMD dogs carried significantly more fecal AMR E. coli (p < 0.001), including third-generation cephalosporin resistant, extended-spectrum beta-lactamase (ESBL) producing, and multidrug resistant isolates and multivariable modeling confirmed raw-meat diets to be a significant risk factor. The bla CTX-M-15 gene was the most frequently identified bla ESBL gene. The bla CTX-M-55 and bla SHV-66 genes were also prevalent and were only found in RMD dogs. The mobile colistin resistance gene, mcr-4 was identified in one ESBL-producing E. coli isolate from a NRMD-fed dog. This study has shown that dogs fed RMD in the UK are significantly more likely to shed E. coli which is resistant to highest priority critically important antibiotics, and multidrug resistant E. coli, than dogs fed NRMD. Additionally, AMR-E. coli isolates from RMD-fed dogs harbor multiple, diverse, and novel AMR genes. Therefore, provision of RMD to dogs could pose an important potential threat to human and animal health, especially given the close nature of the relationship many owners share with their pets. Awareness of these findings should be shared with pet owners, veterinary and medical professionals, pet food manufacturers and public health to mitigate potential risks.

Potential of training of anti-Staphylococcus aureus therapeutic phages against Staphylococcus epidermidis multidrug-resistant isolates is restricted by inter- and intra-sequence type specificity.

Phage therapy appears to be a promising approach to tackle multidrug-resistant bacteria, including staphylococci. However, most anti-staphylococcal phages have been characterized in Staphylococcus aureus, while a limited number of studies investigated phage activity against S. epidermidis. We studied the potential of phage training to extend the host range of two types of anti-S. aureus phages against S. epidermidis isolates. The Appelmans protocol was applied to a mixture of Kayvirus and a mixture of Silviavirus phages repeatedly exposed to seven S. epidermidis strains representative of nosocomial-associated sequence types (ST), including the world-wide disseminated ST2. We observed increased activity only for the Kayvirus mixture against two of these strains (ST2 or ST35). Phage subpopulations isolated from the training mixture using these two strains (five/strain) exhibited different evolved phenotypes, active only against their isolation strain or strains of the same ST. Of note, 16/47 ST2 strains were susceptible to one of the groups of trained phages. A comparative genomic analysis of ancestral and trained phage genomes, conducted to identify potential bacterial determinants of such specific activity, found numerous recombination events between two of the three ancestors. However, a small number of trained phage genes had nucleotide sequence modifications impacting the corresponding protein compared to ancestral phages, two to four of them per phage genome being specific of each group of phage subpopulations exhibiting different host range. The results suggest that anti-S. aureus phages can be adapted to S. epidermidis isolates but with inter- and intra-ST specificity.ImportanceS. epidermidis is increasingly recognized as a threat for public health. Its clinical importance is notably related to multidrug resistance. Phage therapy is one of the most promising alternative therapeutic strategies to antibiotics. Nonetheless, only very few phages active against this bacterial species have been described. In the present study, we showed that phage training can be used to extend the host range of polyvalent Kayvirus phages within the Staphylococcus genera to include S. epidermidis species. In the context of rapid development of phage therapy, in vitro forced adaptation of previously characterized phages could be an appealing alternative to fastidious repeated isolation of new phages to improve the therapeutic potential of a phage collection.

Biofilm formation, antibiotic-resistance and clonal relatedness among clinical isolates of Acinetobacterbaumannii

In this work, the antibiotic resistance, biofilm formation capability, and clonal relatedness of 50 A. baumannii isolates collected from three hospitals in Ardabil city, Iran, were evaluated. Antibiotic sensitivity and biofilm formation of isolates were determined by disk diffusion and microtiter-plate methods, respectively. Molecular typing of isolates was also performed using repetitive sequence-based PCR (REP-PCR). The majority of isolates were resistant to cephems, aminoglycosides, and carbapenems, with 80 % classified as multi-drug resistant (MDR). While, only isolates collected from blood and tracheal were resistant to colistin. Additionally, 42 isolates (84 %) had biofilm formation capability. According to rep-PCR results, 34 isolates showed similar banding patterns, while 16 isolates had unique banding patterns. Finally, based on the molecular analysis, there was a direct relationship between biofilm formation and the antibiotic resistance of isolates. In other words, MDR isolates had a higher ability to form biofilm.

Genome sequences of 70 multidrug-resistant Gram-negative isolates in high-risk neonates in the Northeast of Mexico.

Infections by multidrug-resistant pathogens are steadily increasing worldwide. A considerable proportion of neonatal intensive care admissions have a bacterial infection with multidrug-resistant bacteria during their hospital stay. In this work, we report draft genome sequences of 70 selected isolates from high-risk neonates in the Northeast of Mexico.

Minimum inhibitory and bactericidal/fungicidal concentration of commercially available products containing essential oils, zinc gluconate, or 4% chlorhexidine for Malassezia pachydermatis, Pseudomonas aeruginosa, and multi-drug resistant Staphylococcus pseudintermedius canine clinical isolates.

Skin infections are common complications in both humans and animals. Because of the increased incidence of multi-drug resistant (MDR) skin infections, essential oils have been suggested as potential alternatives to the classic antimicrobials. The goal of this study was to evaluate the minimum inhibitory and bactericidal/fungicidal concentrations (MIC and MBC/MFC) of commercially available products containing essential oils, zinc gluconate, or 4% chlorhexidine. Microbroth dilution technique was performed on clinical isolates of MDR Staphylococcus pseudintermedius (MDR-SP; n = 10), Pseudomonas aeruginosa (PA; n = 10), and Malassezia pachydermatis (MP; n = 10). For MDR-SP, essential oil-containing products showed median MICs of 1:240 and 1:320. The chlorhexidine shampoo had a MIC of 1:128,000 (0.312µg/mL), whereas zinc gluconate products had median MICs of 1:320 and 1:160. Three essential oil-containing shampoos (MBC 1:40), the zinc gluconate (MBC 1:40), and the chlorhexidine (MBC 1:64,000 [0.625µg/mL]) reached an MBC. For PA, essential oil-containing products showed median MICs of 1:30 and 1:80. The zinc-gluconate products had a median MIC of 1:160, whereas the chlorhexidine shampoo had a median MIC of 1:4,000 (10µg/mL). Only the zinc-gluconate products (MBC 1:80) and the chlorhexidine shampoo (MBC 1:2,000 [20µg/mL]) reached an MBC. For MP, essential oil-containing and zinc-gluconate products showed lower median MICs (1:4,800 and 7,200) for shampoos compared with other formulations (1:160 and 1:320), whereas the chlorhexidine shampoo had a median MIC of 1:80,000 (0.5µg/mL). These results suggest that natural topical compounds can be an effective alternative to treat skin infections in companion animals. Further in vivo studies are needed to clinically confirm this study's results.

Perception and Role of Indirect and Post-Discharge Surveillance in Predicting Surgical Site Infection in Surgical Patients: An Institutional Experience

Abstract Background: Surgical site infection (SSI) surveillance is a process of observing and reporting of infections following surgery. SSI surveillance studies are few in our country. Objectives: Present study was conducted to know the perception and role of SSI surveillance. Incidence of surgical site infection, identification of risk factors and bacterial spectrum of surgical wounds during surveillance were also studied. Methods: Patients who underwent a surgical procedure between July 2022 and June 2023 at our hospital were included. A prospective incidence surveillance for SSI was done to look for any signs of infection and record them in the surveillance form. Wound swabs or pus aspirates were sent for microbiological culture. Results: A total of 2312 patients underwent surgery during the study period, of whom 61 patients were diagnosed with SSI. The incidence of SSI was 2.6%. SSI was encountered more in female patients (57.3%). Age distribution had a significant association with SSI (P = 0.025). Most operative wounds were clean-contaminated (45.9%). Most SSIs were superficial (91.8%), followed by deep (8.2%). Most patients who developed SSI had a National Nosocomial Infection Surveillance (NNIS) patient risk index of 2 (45.9%). Bacterial growth was observed in 45.9% of cases. Multidrug resistance (MDR) among these isolates was 42.6%. Conclusion: This study helped to determine predictors of SSI like duration of surgery, type of wound, duration of hospital stays, and NNIS risk index. MDR isolates are now contributing to SSI. The perception of healthcare workers about SSI surveillance was that it was feasible to use and practice. Surveillance helped clinicians to strengthen the existing infection prevention and control practices for the future.

Vibriocidal efficacy of Bifidobacterium bifidum and Lactobacillus acidophilus cell-free supernatants encapsulated in chitosan nanoparticles against multi-drug resistant Vibrio cholerae O1 El Tor

BackgroundCholera is a diarrheal disease recognized for being caused by toxin-producing Vibrio (V.) cholerae. This study aims to assess the vibriocidal and immunomodulatory properties of derived cell-free supernatants (CFSs) of Bifidobacterium (B.) bifidum and Lactobacillus (L.) acidophilus encapsulated in chitosan nanoparticles (CFSb-CsNPs and CFSa-CsNPs) against clinical multi-drug resistance (MDR) isolates of V. cholerae O1 El Tor.MethodsWe synthesized CFSb-CsNPs and CFSa-CsNPs using the ionic gelation technique. The newly nanostructures were characterized for size, surface zeta potential, morphology, encapsulation efficacy (EE), stability in different pH values and temperatures, release profile, and in vitro cytotoxicity. The antimicrobial and antibiofilm effects of the obtained nanocomposites on clinical MDR isolates (N = 5) of V. cholerae E1 Tor O1 were investigated by microbroth dilution assay and crystal violet staining, respectively. We conducted quantitative real-time PCR (qRT-PCR) to analyze the relative gene expressions of Bap, Rbmc, CTXAB, and TCP in response to CFSb-CsNPs and CFSa-CsNPs. Additionally, the immunomodulatory effects of formulated structures on the expression of TLR2 and TLR4 genes in human colorectal adenocarcinoma cells (Caco-2) were studied.ResultsNano-characterization analyses indicated that CFSb-CsNPs and CFSa-CsNPs exhibit spherical shapes under scanning electron microscopy (SEM) imaging, with mean diameters of 98.16 ± 0.763 nm and 83.90 ± 0.854 nm, respectively. Both types of nanoparticles possess positive surface charges. The EE% of CFSb-CsNPs was 77 ± 4.28%, whereas that of CFSa-CsNPs was 62.5 ± 7.33%. Chitosan (Cs) encapsulation leads to increased stability of CFSs in simulated pH conditions of the gastrointestinal tract in which the release rates for CFSb-CsNPs and CFSa-CsNPs were reached at 58.00 ± 1.24% and 55.01 ± 1.73%, respectively at pH = 7.4. The synergistic vibriocidal effects observed from the co-administration of both CFSb-CsNPs and CFSa-CsNPs, as evidenced by a fractional inhibitory concentration (FIC) index of 0.57, resulting in a significantly lower MIC of 2.5 ± 0.05 mg/mL (p < 0.0001) compare to individual administration. The combined antibacterial effect of CFSb-CsNPs and CFSa-CsNPs on Bap (0.14 ± 0.05), Rbmc (0.24 ± 0.01), CTXAB (0.30 ± 0.09), and TCP (0.38 ± 0.01) gene expression was significant (p < 0.001). Furthermore, co-administration of CFSb-CsNPs and CFSa-CsNPs also demonstrated the potency of suppressing TLR 2/4 (0.20 ± 0.01 and 0.12 ± 0.02, respectively) gene expression (p = 0.0019) and reduced Caco-2 cells attached bacteria to 526,000 ± 51,46 colony-forming units/mL (11.19%) (p < 0.0001).ConclusionOur study revealed that encapsulating CFSs within CsNPs enhances their vibriocidal activity by improving stability and enabling a controlled release mechanism at the site of interaction between the host and bacteria. Additionally, the simultaneous use of CFSb-CsNPs and CFSa-CsNPs exhibited superior vibriocidal potency against MDR V. cholerae O1 El Tor strains, indicating these combinations as a potential new approach against MDR bacteria.

The emergence of transposon-driven multidrug resistance in invasive nontypeable Haemophilus influenzae over the last decade

Nontypeable Haemophilus influenzae (NTHi), once considered a harmless commensal, has emerged as a significant concern due to the increased prevalence of multidrug-resistant (MDR) strains and their association with invasive infections. This study aimed to explore the epidemiology and molecular resistance mechanisms of 51 NTHi isolates collected from patients with invasive infections in northern Taiwan between 2011 and 2020. This investigation revealed substantial genetic diversity, encompassing 29 distinct sequence types and 18 clonal complexes. Notably, 68.6% of the isolates exhibited ampicillin resistance, with 28 categorised as MDR and four isolates were even resistant to up to six antibiotic classes. Among the MDR isolates, 18 pulsotypes were identified, indicating diverse genetic lineages. Elucidation of their resistance mechanisms revealed 18 β-lactamase-producing amoxicillin–clavulanate-resistant (BLPACR) isolates, 12 β-lactamase-producing ampicillin-resistant (BLPAR) isolates, and 5 β-lactamase-nonproducing ampicillin-resistant (BLNAR) isolates. PBP3 analysis revealed 22 unique substitutions in BLPACR and BLNAR, potentially contributing to cephem resistance. Notably, novel transposons, Tn7736-Tn7739, which contain critical resistance genes, were discovered. Three strains harboured Tn7739, containing seven resistance genes [aph(3′)-Ia, blaTEM-1, catA, sul2, strA, strB, and tet(B)], while four other strains carried Tn7736, Tn7737, and Tn7738, each containing three resistance genes [blaTEM-1, catA, and tet(B)]. The emergence of these novel transposons underscores the alarming threat posed by highly resistant NTHi strains. Our findings indicated that robust surveillance and comprehensive genomic studies are needed to address this growing public health challenge.

Retrospective analysis of Acinetobacter baumannii bacteraemia risk factors, complications and mortality in a tertiary university hospital in Saudi Arabia.

Introduction. There are many multidrug-resistant isolates of the nosocomial pathogen, Acinetobacter baumannii, causing severe healthcare-acquired infections in terminally ill patients with high mortality and morbidity rates. Aim. This study aims to retrospectively analyse A. baumannii bacteraemia (ABB) cases in Saudi Arabia, where the information is sparse regarding the prevalence, risk factors, clinical disease, antibiotic regimen, antibiotic susceptibility, treatment outcomes and mortality associated with this infection. Methods. A retrospective chart review was conducted between 1 January 2015 and 31 December 2022 to identify all patients aged 14 years and above with ABB. Demographic and clinical data, as well as results from laboratory analyses, were collected from patients' electronic charts. Statistical analyses were performed on the data to identify factors associated with 90-day mortality. Results. Of the 122 ABB cases, 71 (63.4%) died. The factors that were found to be associated with 90-day mortality were the Charlson Comorbidity Index, Pitt bacteraemia score, quick Sequential Organ Failure Assessment score (P<0.001 for each), hospital ward (P<0.02), short duration of antibiotic treatment (P<0.01) and higher age (P<0.05). The most common source of infection was central line-associated bloodstream infection in 52.7%. Also associated with mortality were inappropriate antimicrobial therapy (P<0.02) and empirical use of colistin (P<0.05). In many patients, ABB was caused by carbapenem-resistant A. baumannii [(CRAB), 69.6%], and 74.4% of those patients died. Conclusion. To prevent ABB-associated mortality, an appropriate regimen and duration of treatment are necessary. Hospitals should also practice proper hygiene to prevent the spread of ABB. CRAB is a growing threat in hospitals in Saudi Arabia, especially in the critical care setting, and carries a very high risk of mortality.

Multidrug Resistance Gram-negative Bacteria in Intensive Care Unit of Tertiary Care Hospital: A Descriptive Cross-sectional Study

Introduction: Antimicrobial resistance is global health problem, amongst major causes for mortality. It is one of the hinderance for achievement of Sustainable Goal 3 (Good health and well-being) of WHO. Multidrug resistant gram negative bacteria are major threat to humanity especially patients admitted in intensive care unit. This is associated with to treatment failure and mortality of the patients in intensive care unit. Therefore, this study was conducted to find out the prevalence of Multidrug resistant gram negative bacteria in intensive care unit of tertiary care center. Methods: A descriptive cross-sectional study was conducted in the Department of Microbiology of a tertiary care hospital from February, 2020 till August, 2021 for 18 months after obtaining ethical approval from the Institutional Review Committee (Reference Number: 246). All the samples from Intensive Care Unit were processed following standard methodology. Only Gram-negative bacteria isolated from samples were included in the study. Convenience sampling method was used. The point estimated was calculated at 95% Confidence Interval. Data was entered in Microsoft Excel 2016 and analysis was done using IBM SPSS Statistics version 16.0.Results: Out of 500 samples only 380 showed growth of gram negative bacteria. The prevalence Multi Drug Resistance was 83.15%. The prevalence of multidrug resistant was notably higher among Acinetobacter spp (100%) followed by Non fermenter (95%) and Escherichia coli (87.1%). Multidrug resistant isolates were least resistant towards carbapenem group of antibiotics.Conclusions: The prevalence of Multidrug resistant gram negative bacteria was found to be lower than in the study conducted in similar setting.

The antibiotic resistance profiles of Pseudomonas aeruginosa in the Asia Cornea Society Infectious Keratitis Study

PurposeTo describe the prevalence and antibiotic resistance profiles of Pseudomonasaeruginosa isolated from the Asia Cornea Society Infectious Keratitis Study (ACSIKS).MethodsAll bacterial isolates from ACSIKS underwent repeat microbiological identification in a central repository in Singapore. Minimum inhibitory concentration (MIC) determination was conducted for isolates of P. aeruginosa against thirteen antibiotics from 6 different classes, and categorized based on Clinical Laboratory Standard Institutes’ reference ranges. The percentage rates of resistance (non-susceptibility) to each antibiotic included isolates of both intermediate and complete resistance. Multi-drug resistance (MDR) was defined as non-susceptibility to at least one agent in three or more antimicrobial classes.ResultsOf the 1493 unique bacterial specimens obtained from ACSIKS, 319 isolates were of P. aeruginosa. The majority of isolates were from centers in India (n = 118, 37%), Singapore (n = 90, 28.2%), Hong Kong (n = 31, 9.7%) and Thailand (n = 30, 9.4%). The cumulative antibiotic resistance rate was the greatest for polymyxin B (100%), ciprofloxacin (17.6%) and moxifloxacin (16.9%), and lowest for cefepime (11.6%) and amikacin (13.5%). Isolates from India demonstrated the highest antibiotic resistance rates of all the centers, and included moxifloxacin (47.5%) and ciprofloxacin (39.8%). Forty-eight of the 59 MDR isolates also originated from India. Antibiotic resistance rates were significantly lower in the other ACSIKS centers, and were typically less than 10%.ConclusionsThe antibiotic resistance profiles of P. aeruginosa varied between different countries. While it was low for most countries, substantial antibiotic resistance and a significant number of multi-drug resistant isolates were noted in the centers from India.

Unravelling the complex mechanisms of multidrug resistance in bovine mastitis pathogens: Insights into antimicrobial resistance genes, biofilm dynamics, and efflux systems

Mastitis remains a paramount economic threat to dairy livestock, with antibiotic resistance severely compromising treatment efficacy. This study provides an in-depth investigation into the multidrug resistance (MDR) mechanisms in bacterial isolates from bovine mastitis, emphasizing the roles of antimicrobial resistance genes (ARGs), biofilm formation, and active efflux systems. A total of 162 Staphylococci, eight Escherichia coli, and seven Klebsiella spp. isolates were obtained from 215 milk samples of clinical and subclinical mastitis cases. Antibiotic susceptibility testing identified Twenty Staphylococci (12.35 %), six E. coli (75 %) and seven Klebsiella (100 %) identified as MDR displaying significant resistance to β-lactams and tetracyclines The Multiple Antibiotic Resistance (MAR) index of these isolates ranged from 0.375 to 1.0, highlighting extensive resistance. Notably, 29 of the 33 MDR isolates produced biofilms on Congo red agar, while all exhibited biofilm formation in the Microtitre Plate assay. Critical ARGs (blaZ, blaTEM, blaCTX-M, tetM, tetA, tetB, tetC, strA/B, aadA) and efflux pump genes (acrB, acrE, acrF, emrB, norB) regulating active efflux were identified. This pioneering study elucidates the synergistic contribution of ARGs, biofilm production, and efflux pump activity to MDR in bovine mastitis pathogens. To our knowledge, this comprehensive study is the first of its kind, offering novel insights into the complex resistance mechanisms. The findings underscore the imperative need for advanced antibiotic stewardship and strategic interventions in dairy farming to curb the rise of antibiotic-resistant infections, thereby protecting both animal and public health.

Green synthesized silver nanoparticles from Ocimum sanctum: A potent inhibitor of biofilm forming ability and efflux pumps in bacteria causing bovine mastitis

Therapeutic management of mastitis faces significant challenges due to multidrug resistance. In the present study, multi-drug-resistant (MDR) Staphylococcus spp, Klebsiella pneumoniae, and Escherichia coli were isolated from bovine clinical mastitis cases and the phenotypic and genotypic multidrug resistance profiling was carried out. Silver nanoparticles (AgNPs) were biosynthesized using Ocimum sanctum leaf extracts and characterized via UV Vis absorption, Fourier Transform Infrared Spectroscopy, X-ray diffraction studies, Energy dispersive spectroscopy and Electron Microscopy. The determined minimum inhibitory concentration and minimum bactericidal concentration of the AgNPs against the recovered MDR isolates were 62.5 μg/ml and 125 μg/ml respectively. At a concentration of 50 μg/ml, the AgNPs demonstrated biofilm inhibitory activities of 80.35 % for MDR E. coli, 71.29 % for S. aureus and 60.18 % for MDR K. pneumoniae. Post-treatment observations revealed notable differences in biofilm formation across bacterial isolates. Furthermore, AgNP treatment led to significant downregulation of expression of the efflux pump genes acrB, acrE, acrF, and emrB in Gram-negative isolates and norB in Staphylococci isolates. This research underscores the potential for the development of an eco-friendly antimicrobial alternative in the form of green synthesized silver nanoparticles to combat drug resistance offering potential antibiofilm and efflux pump inhibitory activities.

Antibiotic Susceptibility of Bacteria Isolated from Livestock-exposed and Unexposed Areas in Perlite-rich Soil in Thailand

Antibiotic resistant bacteria are present in various environments and are concerning for public health. Antibiotic resistance genes in soil may proliferate throughout different ecosystems and be acquired by pathogens that pose health risks to humans and animals. This study investigated the antibiotic resistance of cultivable bacteria isolated from livestock-exposed and unexposed areas in a former perlite mining area in the Fa-La-Mee Mountain range, Lopburi, Thailand. Soil samples were collected in five different locations. Soil bacteria were isolated and tested for susceptibility against seven antibiotics, followed by 16S rRNA gene-based identification. A total of 56 bacterial isolates were isolated from the perlite-rich soil samples, including 34 isolates from the livestock-exposed areas and 22 isolates from the unexposed areas. Most of the isolates were resistant to cefepime (26.5% in exposed areas and 22.7% in unexposed areas) and ampicillin (23.5% in exposed areas and 22.7% in unexposed areas). 16S rRNA gene sequences revealed that most of the resistant bacteria isolated from perlite-rich soil were Bacillus spp. A multidrug-resistant isolate of Bacillus cereus resistant to ampicillin, trimethoprim/sulfamethoxazole, and cefepime was found in a livestock-unexposed area. The majority of the culturable antibiotic-resistant bacteria isolated from perlite-rich soil were cefepime-resistant Bacillus. Interestingly, the pattern of antibiotic resistance in exposed and unexposed areas was not different, which indicated that antibiotic resistance in perlite-rich soil is likely caused by other factors, such as physicochemical characteristics and/or the microbial population of the soil rather than by antibiotics used in livestock husbandry.

Colorimetric Loop-Mediated Isothermal Amplification Assays Accurately Detect blaOXA-23-like and ISAba1 Genes from Acinetobacter baumannii in Pure Cultures and Spiked Human Sera.

Carbapenem resistance in Acinetobacter baumannii is a critical global health threat attributed to transferrable carbapenemase genes. Carbapenemase genotyping using polymerase chain reaction (PCR) presents a challenge in resource-limited settings because of its technical requirements. This study designed new loop-mediated isothermal amplification (LAMP) primers using multiple sequence alignment-based workflows, validated the primer performance against multiple target variants in silico, and developed novel LAMP assays (LAntRN-OXA23 and LAntRN-ISAba1) to detect the transferable blaOXA-23-like carbapenemase genes and ISAba1 elements in pure cultures and A. baumannii-spiked serum samples. The designed LAMP primers bind to the conserved regions of their highly polymorphic targets, with their in silico performance comparable with other published primers. The in vitro LAMP assays (using 30 PCR-profiled A. baumannii and 10 standard multidrug-resistant gram-negative isolates) have 100% concordance with the PCR-positive clinical samples, limits of detection as low as 1 pg/µL (200 copies/µL), and specificities of 57.89-100%. Both assays produced positive results when testing DNA samples (extracted using a commercial kit) from blaOXA-23-like and ISAba1-blaOXA-51-like PCR-positive A. baumannii-spiked normal human sera (five set-ups per target). In summary, the LAMP assays accurately detected the target genes and have applications in infection management, control, and point-of-care testing in resource-limited healthcare settings.

Discovery and Biosynthesis of Persiathiacins: Unusual Polyglycosylated Thiopeptides Active Against Multidrug Resistant Tuberculosis.

Thiopeptides are ribosomally biosynthesized and post-translationally modified peptides (RiPPs) that potently inhibit the growth of Gram-positive bacteria by targeting multiple steps in protein biosynthesis. The poor pharmacological properties of thiopeptides, particularly their low aqueous solubility, has hindered their development into clinically useful antibiotics. Antimicrobial activity screens of a library of Actinomycetota extracts led to discovery of the novel polyglycosylated thiopeptides persiathiacins A and B from Actinokineospora sp. UTMC 2448. Persiathiacin A is active against methicillin-resistant Staphylococcus aureus and several Mycobacterium tuberculosis strains, including drug-resistant and multidrug-resistant clinical isolates, and does not significantly affect the growth of ovarian cancer cells at concentrations up to 400 μM. Polyglycosylated thiopeptides are extremely rare and nothing is known about their biosynthesis. Sequencing and analysis of the Actinokineospora sp. UTMC 2448 genome enabled identification of the putative persiathiacin biosynthetic gene cluster (BGC). A cytochrome P450 encoded by this gene cluster catalyzes the hydroxylation of nosiheptide in vitro and in vivo, consistent with the proposal that the cluster directs persiathiacin biosynthesis. Several genes in the cluster encode homologues of enzymes known to catalyze the assembly and attachment of deoxysugars during the biosynthesis of other classes of glycosylated natural products. One of these encodes a glycosyl transferase that was shown to catalyze attachment of a D-glucose residue to nosiheptide in vitro. The discovery of the persiathiacins and their BGC thus provides the basis for the development of biosynthetic engineering approaches to the creation of novel (poly)glycosylated thiopeptide derivatives with enhanced pharmacological properties.

Design, Synthesis, and Anticancer and Antibacterial Activities of Quinoline-5-Sulfonamides.

A series of new unique acetylene derivatives of 8-hydroxy- and 8-methoxyquinoline- 5-sulfonamide 3a-f and 6a-f were prepared by reactions of 8-hydroxy- and 8-methoxyquinoline- 5-sulfonyl chlorides with acetylene derivatives of amine. A series of new hybrid systems containing quinoline and 1,2,3-triazole systems 7a-h were obtained by reactions of acetylene derivatives of quinoline-5-sulfonamide 6a-d with organic azides. The structures of the obtained compounds were confirmed by 1H and 13C NMR spectroscopy and HR-MS spectrometry. The obtained quinoline derivatives 3a-f and 6a-f and 1,2,3-triazole derivatives 7a-h were tested for their anticancer and antimicrobial activity. Human amelanotic melanoma cells (C-32), human breast adenocarcinoma cells (MDA-MB-231), and human lung adenocarcinoma cells (A549) were selected as tested cancer lines, while cytotoxicity was investigated on normal human dermal fibroblasts (HFF-1). All the compounds were also tested against reference strains Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212 and representatives of multidrug-resistant clinical isolates of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant E. faecalis. Only the acetylene derivatives of 8-hydroxyquinoline-5-sulfonamide 3a-f were shown to be biologically active, and 8-hydroxy-N-methyl-N-(prop-2-yn-1-yl)quinoline-5-sulfonamide (3c) showed the highest activity against all three cancer lines and MRSA isolates. Its efficacies were comparable to those of cisplatin/doxorubicin and oxacillin/ciprofloxacin. In the non-cancer HFF-1 line, the compound showed no toxicity up to an IC50 of 100 µM. In additional tests, compound 3c decreased the expression of H3, increased the transcriptional activity of cell cycle regulators (P53 and P21 proteins), and altered the expression of BCL-2 and BAX genes in all cancer lines. The unsubstituted phenolic group at position 8 of the quinoline is the key structural fragment necessary for biological activity.

Comparison of Beta-Lactamase Profiles and Epidemiology of Multi-Drug Resistant Acinetobacter baumannii Isolates in Southwest Turkey: 2011 vs. 2019 Study

Background: The most significant problem encountered with hospital infections caused by multi-drug resistant Acinetobacter baumannii isolates is the difficulty in treatment. Defining resistance mechanisms is important for the development of treatment strategies. Objectives: The aim of this study was to investigate the beta-lactamase genes and clonal relationship in multi-drug resistant A. baumannii isolates in Mugla, Turkey, as no data are currently available for this province. Methods: Total 96 multi-drug resistant A. baumannii isolates obtained from hospitalized patients, between 2011 - 2012 period (n = 61) and in year 2019 (n = 35) were investigated and compared for antibiotic susceptibilities, beta-lactamase enzyme encoding genes by polymerase chain reaction (PCR) method. Repetitive extragenic palindromic (REP) PCR was made for dendrogram analysis to illustrate hierarchical relationship between isolates. Results: Overall, 74.0% of the isolates were obtained from patients in intensive care units. All multi-drug resistant isolates were found resistant to ciprofloxacin, piperacillin, piperacillin-tazobactam, meropenem, and imipenem by the disk diffusion method. The minimum inhibitory concentration values indicated that all isolates were resistant to imipenem, while colistin resistance was determined as 21.9%. The ISAba-I insertion element was present in all isolates. BlaOXA-23-like was present in 96.9%, and blaOXA-24-like was detected in three (3.1%) isolates. blaNDM positivity was shown in 5.2% of isolates. The indistinguishable or closely related isolates ratio was detected as 29.5% for the 2011 - 2012 period and 80% for 2019. Conclusions: Our findings showed that antibiotic resistance rates were high in A. baumannii, particularly isolated from intensive care unit patients in Mugla province. The prevalence of beta-lactamase and metallo-beta-lactamase genes has been increasing year by year, with the blaOXA-23-like gene region being the most prevalent. Additionally, the co-existence of these genes among strains is also on the rise. These findings from two different periods may provide useful reference data for future studies.