Beta-lactamase Research Articles

Integrating molecular docking and molecular dynamics simulation studies on the affinity and interactions of piperine with β-lactamase class A enzymes

ContextOne of the major problems facing health systems is the emergence of resistant bacteria. Emerge of extended spectrum of bacterial generated β-lactamases, due to genetic variation, has reduced the effectiveness of β-lactam antibiotics. Hydrolysis of the rings in the structure of β-lactam antibiotics that performed by β-lactamases lead to Bacterial resistance. It is necessary to discover compounds with beta-lactamase inhibitory properties. This work proposes piperine which is the main components of the black pepper and has several medicinal activities can be used as inhibitor of β-lactamase. Molecular docking results show that piperine bounded to all three β-lactamase class A enzymes and has suitable binding energies ranging from −6.43 to −6.66 kcal/mol. Analysis of the MD simulation trajectories predicted the formation of stable hydrogen bonds between piperine and all three β-lactamase class A enzymes during the whole simulation time which confirmed the molecular docking results. MethodsMolecular docking were performed to determine the binding pose and binding energy of class A beta lactamase with piperine using Autodock 4.2.2 software. Then the lowest conformation in the high populated cluster chosen for further analysis. Finally molecular dynamic simulation was carried out for three enzymes utilizing GROMACS 2019.6 program applying AMBER99SB force field.

Screening for Beta-Lactamases Producing Bacteria and Other Resistant Bacteria among Patients Admitted to the Emergency Unit at the Yaoundé University Teaching Hospital, Cameroon

Aims: This study aimed to screen for Beta Lactamases (BLs) and other resistant bacteria among patients admitted to the emergency unit of the Yaoundé University Teaching Hospital (YUTH).
 Study Design: This study was a cross-sectional hospital-based study.
 Place and Duration of Study: This study was conducted at the Bacteriology unit of the YUTH, Yaoundé- Cameroon between February to June 2020.
 Methodology: Seventy-Five urine samples were collected from newly admitted patients at the emergency unit of the YUTH and bacteria species were identified basis on their culture characteristics, Gram morphology, and biochemical tests. The isolates were screened from the production of extended-spectrum beta-lactamases (ESBLs) and AmpC BLs using the Double disk synergy method and Disk approximation methods respectively. These isolates were later subjected to antimicrobial susceptibility testing using the disk diffusion method.
 Results: Out of the 75 urine analyzed, 14 (18.7%) were found positive for Urinary Tract Infection. Fourteen bacteria species were isolated identified and enumerated as E. coli (5), Klebsiella species (4), Citrobacter species (2), Proteus species (2), and Enterobacter species (1). A high level of resistance was observed with Amoxicillin clavulanic acid, Cefuroxime, and Ceftazidime while a high level of sensitivity was observed among carbapenem antibiotics. Eight of 14 isolated bacteria were BLs producers, of which 5 were sorely ESBL producers, 2 co-producers (ESBL + AmpC), and 1 AmpC producer. The overall positive rate of BLs in the study population was 10.7%. Again, patient origin and previous antibiotic use were significantly associated with BLs prevalence p-value of .01 and .04 respectively.
 Conclusion: The high prevalence of the ß- lactamases in the Emergency unit emphasizes the need for continuous surveillance in the Emergency unit to detect resistant strains, strict guidelines for antibiotic therapy, and the implementation of infection control measures to reduce the increasing burden of antibiotic resistance.

Isolation and Speciation of Acinetobacter Species with Special Reference to Antibiotic Resistance in Tertiary Care Hospital

Acinetobacter can survive longer in the environment and is also known for developing resistance against agents like disinfectants and nutritional deprivation. Very restricted information about Acinetobacter is available because of their confused taxonomic status. The isolation and identification of resistance pattern help in the selection and search for new antibiotics, reducing the morbidity and mortality of patients. The present study was conducted to find the different types of resistant mechanisms in Acinetobacter species and their antimicrobial susceptibility pattern from various clinical samples. Gram's stain and biochemical reactions identified Acinetobacter isolated from various clinical samples. Antibiotic susceptibility testing was done, and their resistant pattern was observed. Phenotypic methods for drug resistance were carried out -Detection of Extended Spectrum Beta Lactamase (ESBL) by Double disc synergy test (DDST), detection of Metallo Beta Lactamase (MBL) by Imipenem with EDTA, and detection of Carbapenemase production by Modified Hodge test (MHT). The results showed that out of 150 isolates of Acinetobacter species, Acinetobacter baumannii 138 (92%) was the most common species isolated, followed by Acinetobacter lwoffii 10 (7%) and Acinetobacter hemolytic 2 (1%). Of these, 22 (15%) isolates showed ESBL production by Double disc synergy test, 80 (53%) isolates were MBL producers, and 11 (7%) were Carbapenemase producers by Modified Hodge test. The study observed that ESBL production in Acinetobacter was 22(15%) and MBL 80(53%). It demonstrated that most of the Acinetobacter isolated were found to be Multi-Drug Resistant (95%). It brings out the need for active surveillance combined to eradicate the curtail of this organism in hospital settings.

Exploration of the protein-dependent mechanism of Lactobacillus crispatus GAI98322 to prevent recurrent cystitis

ObjectivesTo elucidate the mechanism of Lactobacillus crispatus (L. crispatus) suppositories to prevent patients from recurrent cystitis (RC), independent from viable-Lactobacilli-bacteria- and acid-dependent ones such as hydrogen peroxide and lactate. MethodsWe used the GAI98322 strain of L. crispatus in all experiments and pH-matched.cell-free culture supernatant of L. crispatus (CFCS) was collected. The growth inhibitory activity and the biofilm formation inhibitory activity of the CFCS against uropathogenic Escherichia coli (UPEC), Extended Spectrum beta (β) Lactamase producing (ESBL+) UPEC, and Pseudomonas aeruginosa (P. aeruginosa) was assessed by agar-disk diffusion tests and crystal violet assay. Also, CFCS was subjected to mass spectrometry to specify ingredients. ResultsThe CFCS suppressed the proliferation of E. coli, ESBL + E. coli, and P. aeruginosa. Also, the CFCS at a concentration of 40% significantly impeded the biofilm formation of these three bacteria. The aggregation-promoting factor and Lysin was detected from CFCS. ConclusionsThe cell-free supernatant from the GAI98322 strain of L. crispatus inhibits the growth/biofilm formation of broad pathogens by aggregation promoting factor and lysin, which may prevent hosts from RC regardless of the antimicrobial resistance of the pathogens and even under pH modulation.

Defining an optimal control for RNAi experiments with adult Schistosoma mansoni

In parasites such as Schistosoma mansoni, gene knockdown by RNA interference (RNAi) has become an indispensable tool for functional gene characterization. To distinguish target-specific RNAi effects versus off-target effects, controls are essential. To date, however, there is still no general agreement about suitable RNAi controls, which limits the comparability between studies. To address this point, we investigated three selected dsRNAs for their suitability as RNAi controls in experiments with adult S.mansoni in vitro. Two dsRNAs were of bacterial origin, the neomycin resistance gene (neoR) and the ampicillin resistance gene (ampR). The third one, the green fluorescent protein gene (gfp), originated from jellyfish. Following dsRNA application, we analyzed physiological parameters like pairing stability, motility, and egg production as well as morphological integrity. Furthermore, using RT-qPCR we evaluated the potential of the used dsRNAs to influence transcript patterns of off-target genes, which had been predicted by si-Fi (siRNA-Finder). At the physiological and morphological levels, we observed no obvious changes in the dsRNA treatment groups compared to an untreated control. However, we detected remarkable differences at the transcript level of gene expression. Amongst the three tested candidates, we suggest dsRNA of the E. coli ampR gene as the most suitable RNAi control.

Factors influencing infections caused by Extended Spectrum Beta Lactamase (ESBL) producing bacteria in patients with urinary tract infections at RSUD dr. R. Soetijono Blora

Urinary tract infection (UTI) is a bacterial infection that is often found in pediatric patients. The prevalence of infections due to ESBL-producing Enterobacterales is increasing worldwide. This study aims to identify and analyze the factors that influence Extended Spectrum Beta Lactamase (ESBL) producing bacteria in UTI patients at RSUD dr. R. Soetijono Blora. A cross-sectional study has been carried out at RSUD dr. R. Soetijono Blora in January-March 2023. The results of this study showed that 58 respondents with urine culture results found Enterobacterales pathogenic bacteria. Based on the sensitivity test for the Extended Spectrum Beta Lactamase (ESBL) enzyme producer, 39 people (67.2%) were categorized as ESBLs and 19 people (32.8%) Non-ESBLs. The study variables included age, gender, history of catheter insertion, history of stones or urinary tract surgery and history of taking antibiotics in the last 6 months. The chi-square test to determine the relationship between variables was used to determine significant risk factors. Bivariate logistic regression showed that history of catheter insertion and history of use of antibiotics in the last 6 months were significant risk factors. Prevalence of UTIs due to ESBL-producing Enterobacterales in RSUD dr. R. Soetijono Blora is high and the wise use of antibiotics, especially the third generation cephalosporins as therapy, is needed for preventive strategies to reduce antibiotic resistance

Acquisition and carriage of genetically diverse multi-drug resistant gram-negative bacilli in hospitalised newborns in The Gambia

BackgroundThis detailed genomic study characterised multi-drug resistant-Gram negative bacilli (MDR-GNB) carriage in neonates < 2 kg and paired mothers at a low-resource African hospital.MethodsThis cross-sectional cohort study was conducted at the neonatal referral unit in The Gambia with weekly neonatal skin and peri-anal sampling and paired maternal recto-vaginal swabs. Prospective bacteriological culture used MacConkey agar with species identification by API20E and API20NE. All GNB isolates underwent whole genome sequencing on Illumina Miseq platform. Multi-Locus Sequence Typing and SNP-distance analysis identified strain type and relatedness.Results135 swabs from 34 neonates and 21 paired mothers, yielded 137 GNB isolates, of which 112 are high quality de novo assemblies. Neonatal MDR-GNB carriage prevalence is 41% (14/34) at admission with 85% (11/13) new acquisition by 7d. Multiple MDR and ESBL-GNB species are carried at different timepoints, most frequently K. pneumoniae and E. coli, with heterogeneous strain diversity and no evidence of clonality. 111 distinct antibiotic resistance genes are mostly beta lactamases (Bla-AMPH, Bla-PBP, CTX-M-15, Bla-TEM-105). 76% (16/21) and 62% (13/21) of mothers have recto-vaginal carriage of ≥1 MDR-GNB and ESBL-GNB respectively, mostly MDR-E. coli (76%, 16/21) and MDR-K. pneumoniae (24%, 5/21). Of 21 newborn-mother dyads, only one have genetically identical isolates (E. coli ST131 and K. pneumoniae ST3476).ConclusionsGambian hospitalised neonates exhibit high MDR and ESBL-GNB carriage prevalence with acquisition between birth and 7d with limited evidence supporting mother to neonate transmission. Genomic studies in similar settings are required to further understand transmission and inform targeted surveillance and infection prevention policies.

Coexistence Of Blaoxa-48, Blavim, And Blashv Genes In Klebsiella Pneumoniae And Escherichia Coli Isolated From Urinary Tract Infections: Microbiological And Epidemiological Analysis.

To investigate antimicrobial resistance mechanisms of isolated bacterialstrains, and their correlation with virulence profile. The cross-sectional study was conducted in January 2020 at outpatient health centres in Kafrelsheikh Governorate of Egypt, and comprised urine samples from patients regardless of age and gender. Midstream samples were collected into sterile swaps which were kept in ice-cooled boxes until transported to the laboratory within 5h. Antimicrobial resistance profile of the isolated Enterobacteriaceae was done using Kirby-Bauer disk diffusion method and was confirmed withVitek compact 2. The phenotypic of carbapenemases and extended-spectrum beta lactamase was determined, and polymerase chain reaction was used, as appropriate. Data was analysed using SPSS 20. Of the 199 patients, 101(50.7%) were females and 98(49.3%) were males. The majority 73(36.6%) were aged 30-50 years. Urinary tract infection was found in 68(34.2%) patients. In 28(41.2%) of these patients, there were 32 isolates of Enterobacterales; 21(65.62%) Klebsiella pneumoniae, 7(21.87%) Escherichia coli and 4(12.5%) Enterobacter cloacae. Of the 28(41.2%) patients, 24(85.7%) were infected with a single strain; 17(70.8%) Klebsiella pneumoniae, 4(16.7%) Escherichia coli and 3(12.5%) Enterobacter cloacae. In 3(10.7%) cases, there was co-infection with Escherichia coli and Klebsiella pneumoniae, and 1(3.6%)sample had mixed infection with Klebsiella pneumoniae and Enterobacter cloacae. The other 40(58.8%) patients had other causative agents. Housewives, agricultural workers and those aged >50 years had a higher risk of urinary tract infections(p<0.05) Among Klebsiella pneumonia isolates, 6(28.5%) possessed carbapenemase-related genes and 4(19.1%) extended-spectrum beta lactamase-related genes. The carbapenemase related genes were bla-Verona integron-encoded metallo beta lactamase 6(100%) bla-New Delhi metallo beta lactamase-1 4(66.6%) and bla-oxacillinase-48 2(33.3%). The 4(19.1%) cases of extended-spectrum beta lactamase related genes had bla-temoneira gene 3(75%) and bla-sulfhydryl variable gene 4(100%). In Escherichia coli isolates, bla-oxacillinase-48 and bla-Cefotaximase genes were observed in 2(28.5%) cases.Virulence genes uridine diphosphate glucose 4-epimerase, fimbrial adhesion and mannose-resistance adhesin of Klebsiella spp genes in Klebsiella pneumoniae isolates were positive in in 16(76.2%), 14(66.7%) and 10(47.6%) cases, respectively. All 21(100%) isolates of Klebsiella pneumoniae were negative for mucoviscosity-associated gene A. There was evidence of the coexistence of bla- oxacillinase-48, bla-Verona integron-encoded metallo beta lactamase and bla-sulfhydryl variable genes in Klebsiella pneumoniae and Escherichia coli isolates from mixed urinary tract infection samples.

In Vitro Study of Antibacterial Activity of Snake Fruit Extract against Extended Spectrum Beta Lactamase (ESBL) Escherichia coli

In Vitro Study of Antibacterial Activity of Snake Fruit Extract against Extended Spectrum Beta Lactamase (ESBL) Escherichia coli

Antibacterial effect of kitchen herbs against pathogenic multidrug-resistant E. coli isolates from calf diarrhoea.

Calf diarrhoea remains the biggest challenge both in the small and large farms. Infectious diarrhoea is associated with many pathogens, Escherichia coli being one, but majority are systematically treated with antibiotics. Since antimicrobial resistance (AMR) is a growing menace, the need to find alternative prophylactic solutions using popular kitchen herbs such as Trachyspermum ammi (carom seeds), Curcuma longa (turmeric) and cinnamon (Cinnamomum sp.) extracts is been investigated against virulent form of E. coli isolated from calf diarrhoea. The virulence factors identified in these isolates were ST (32.5%), LT (20%), eaeA (15%), stx1 (2.5%) and stx2 (5%) with the occurrence of the most common serogroups as O18 (15%) followed by O111 (12.5%). Highest resistance was seen with beta lactam + beta lactamase inhibitor (amoxicillin/clavulanic acid) followed by beta lactams (ampicillin, cefuroxime and cefepime). The zone of inhibition due to cinnamon (methanol) and carom seed (ethanol) extracts (500 to 250 μg/mL concentration) on E. coli bacteria was >19 mm, respectively. Turmeric, cinnamon and carom had the potency of inhibiting the pathogenic E. coli which maybe suggestive of its use in calf diets as prophylaxis against diarrhoea.

Plasmid and Total Protein Analyses of Extended Spectrum Beta Lactamase Producing Bacteria from Periplaneta americana (Coackroaches)

Background: The importance of plasmid-encoded Extended Spectrum Beta Lactamase (ESBL) organisms in the dissemination of multi-drug resistance and complicated infections are contributory factors to treatment failure. Despite these, there is a paucity of information regarding plasmid-borne associated infection from primary sources including cockroaches while studies correlating total protein and plasmid-encoded ESBL are also limited especially in cockroaches. The objective of this study was therefore aimed at determining the presence of plasmid in the ESBL organisms from cockroaches as well as their total protein profiles.&#x0D; Materials and Methods: Bacterial isolation, characterization and molecular identification were carried out in our previous work following standard recommended techniques. These molecularly identified organisms which were delineated into twenty-two (22) were further subjected to plasmid and total protein analyses using alkaline lysis and Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) respectively.&#x0D; Results: Results obtained identifies 18(81.8%) of the ESBL isolates to be harbouring plasmids of different molecular weights (2-21.5kbp). The total protein profile of the plasmid-encoded ESBL and the non-plasmid encoded ESBL reveals differential protein expression patterns except for two non- plasmid encoded ESBL isolates that have similar patterns as that of plasmid-encoded ESBL organisms (isolates 2 and 3).&#x0D; Conclusion: There is a need to monitor the primary sources of infection in the epidemiological distribution of resistance. Improving environmental sanitation through proper disinfection of carrier insects may be a means to curtailing such spread.

Phenotypic and genetic screening of Klebsiella pneumoniae isolates from human UTI patients for beta-lactamases and their genetic diversity analysis by ERIC and REP PCRs.

Klebsiella pneumoniae is one of the major nosocomial pathogens responsible for pneumoniae, septicaemia, liver abscesses, and urinary tract infections. Coordinated efforts by antibiotic stewardship and clinicians are underway to curtail the emergence of antibiotic-resistant strains. The objective of the present study is to characterize K. pneumoniae strains through antibiotic resistance screening for production of beta-lactamases (β-lactamases) such as extended spectrum beta lactamases (ESBLs), AmpC β-lactamases, and carbapenemases by phenotypic and genotypic methods and genetic fingerprinting by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and repetitive element palindromic PCR (REP-PCR). A total of 85 K. pneumoniae strains isolated from 504 human urinary tract infections (UTI) were used in this study. Only 76 isolates showed positive in phenotypic screening test (PST), while combination disc method (CDM) as phenotypic confirmatory test (PCT) confirmed 72 isolates as ESBL producers. One or more β-lactamase genes were detected by PCR in 66 isolates (91.66%, 66/72) with blaTEM gene being the most predominant (75.75%, 50/66). AmpC genes could be detected in 21 isolates (31.8%, 21/66) with FOX gene being the predominant (24.24%, 16/66), whereas NDM-I was detected in a single strain (1.51%, 1/66). Genetic fingerprinting using ERIC-PCR and REP-PCR revealed wide heterogeneity among β-lactamase producing isolates with discriminatory power of 0.9995 and 1, respectively.

Fluoroquinolone should be reconsidered as empirical therapy for adult inpatient urinary tract infections in tertiary hospital: a perspective descriptive study from the microbiological and cost-effectiveness aspects

Background: Urinary Tract Infections (UTI) has become difficult to treat because of its increasing resistance characteristics to antimicrobial agents, especially to fluoroquinolone. The objective of this study was to describe the effectiveness of fluoroquinolone as empirical therapy for adult inpatient UTI. Methods: This descriptive cross-sectional study was performed at Prof. Dr. I.G.N.G. Ngoerah Hospital, a tertiary Hospital in Denpasar, Bali, from January 2020 until March 2020. All urine specimens were examined using bioMérieux VITEK® 2 System. Result: A total of 155 urine samples met the inclusion and exclusion criteria. We found Escherichia coli (39.4%) as the most common microbes followed by Klebsiella pneumoniae (12.3%). Most of the isolates are multi drug resistant organisms (MDRO) (52.9%) and 57% (49) of the Enterobacteriaceae isolates are extended spectrum beta lactamases (ESBLs). The average duration of the empirical antibiotic therapy was 3.7 days for all fluoroquinolone. Ciprofloxacin (53.5%) is the most common empirical therapy, followed by levofloxacin (16.8%). Antimicrobial sensitivity tests showed that bacteria remained highly sensitive to amikacin (96%) and meropenem (94%). The sensitivity test for ciprofloxacin and levofloxacin was only 26% and 4% respectively. The suitability antibiotic results of fluoroquinolone as empirical therapy were only 20% from 110 isolates. The cost ineffectiveness of fluoroquinolone as empirical therapy is Rp.8,402,400 for 3 months. Conclusion: We concluded that both fluoroquinolone (ciprofloxacin and levofloxacin) have a very low sensitivity rate and are not cost-effective, therefore the use of those antimicrobial agents as empirical therapy should be reconsidered.

Phylogenetic analysis of zoonotic Acinetobacter spp. isolated from Geoffroy's bat (Myotis emarginatus), Northern Iraq

The current study was conducted to isolate and evaluate the phylogenetic status of the zoonotic Acinetobacter spp. from the Geoffroy's bat (Myotis emarginatus). The sample collection included 35 bats captured in some caves in Northern Iraq. Intestine parts and swabs were taken from each bat. The specimens were subjected to bacterial cultivation processes and 16S rRNA gene and beta lactamase (bla TEM-1) dependent polymerase chain reaction (PCR) methods to detect the genus level and the virulence activity, respectively. The phylogenetic tree was built utilizing the 16S rRNA gene. The findings of the bacterial cultivation revealed the presence of the bacterium in 23 (65.7%) of the collected bats; however, the 16S rRNA-PCR showed that only 10 (28.57%) of the bats demonstrated the incidence of this microorganism in their intestines. The bla TEM-1-PCR reported that 4 (40%) isolates of the 16S rRNA-PCR positive bats carried the β-lactamase gene in their genetic materials. The phylogenetic tree showed that the genetic similarity of the current study isolates was closely related to those from Egypt and China. The present data show that Acinetobacter spp. is present in the intestine of the Geoffroy's bat (Myotis emarginatus) located in some caves from Northern Iraq, and some isolates have virulence potential represented by the composition of the beta lactamase gene.

Phenotypic and genetic extended spectrum beta lactamase profiles of bacterial isolates from ICU in tertiary level hospital in Kenya

Background: Bacterial infections in the ICUs are a threat to the lives of critically ill patients. Moreover, their vulnerable immunity predisposes them to developing bacteria-associated sepsis, further deteriorating their already fragile health. In the face of increasing drug resistance menace, the problem of bacterial infection in ICU is likely to worsen. This study sought to assess bacterial infections in ICU setting by identifying prevalent gram-negative bacterial species and characterizing their antibiotic susceptibility patterns. Phenotypic and genetic resistance were determined among gram-negative isolates from ICU patients. Methods: Cross-sectional samples collected from Kenyatta National Hospital ICU between January and June 2021 were cultured for subsequent analysis. Phenotypic identification of culture-positive samples was performed using VITEK 2 while Antibiotic susceptibility patterns were determined based on Antimicrobial Susceptibility Testing (AST) results. Cephalosporin-resistant gram-negative bacteria were assessed by PCR to detect the presence of ESBL genes. Results and discussion: Out of the 168 gram-negative isolates, Acinetobacter baumanii was the most abundant (35%). Other isolates that were present at frequencies more than 15% are Klebsiella pneumoniae and Escherichia. coli. A. baumaniii is known to be a notorious bacterium in ICU due to its multidrug resistance nature. Indeed, A. baumanii isolates from Kenyatta National Hospital showed significantly high level of phenotypic resistance. Concordant with the high level of phenotypic resistance, we found high carriage of the ESBL genes among the isolates analysed in this study. Moreover, majority of isolates harboured all the four ESBL genes. Conclusion: A high rate of phenotypic and genetic resistance was detected among the tested isolates. Resistance to cephalosporins was primarily driven by acquisition of the ESBL genes. The high-rate multidrug resistance in ICU bacterial isolates in this study has a crucial implication for ICU patient management and general antibiotics use.

Drivers of Resistance in Uganda and Malawi (DRUM): a protocol for the evaluation of One-Health drivers of Extended Spectrum Beta Lactamase (ESBL) resistance in Low-Middle Income Countries (LMICs).

In sub-Saharan Africa (sSA), there is high morbidity and mortality from severe bacterial infection and this is compounded by antimicrobial resistance, in particular, resistance to 3rd-generation cephalosporins. This resistance is typically mediated by extended-spectrum beta lactamases (ESBLs). To interrupt ESBL transmission it will be important to investigate how human behaviour, water, sanitation, and hygiene (WASH) practices, environmental contamination, and antibiotic usage in both urban and rural settings interact to contribute to transmission of ESBL E. coli and ESBL K. pneumoniae between humans, animals, and the environment. Here we present the protocol for the Drivers of Resistance in Uganda and Malawi (DRUM) Consortium, in which we will collect demographic, geospatial, clinical, animal husbandry and WASH data from a total of 400 households in Uganda and Malawi. Longitudinal human, animal and environmental sampling at each household will be used to isolate ESBL E. coli and ESBL K. pneumoniae. This will be complimented by a Risks, Attitudes, Norms, Abilities and Self-Regulation (RANAS) survey and structured observations to understand the contextual and psychosocial drivers of regional WASH practices. Bacterial isolates and plate sweeps will be further characterised using a mixture of short-,long-read and metagenomic whole-genome sequencing. These datasets will be integrated into agent-based models to describe the transmission of EBSL resistance in Uganda and Malawi and allow us to inform the design of interventions for interrupting transmission of ESBL-bacteria.

Resistance genes and plasmid profile of bacteria Isolated from selected water bodies in Owo Metropolis

Aquatic samples from three water bodies were collected following laid down procedures. The bacteria were isolated using, Eosin Methylene Blue agar (EMB), Mannitol salt agar (MSA), Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS) Agar, Muller Hinton (MH). The isolates were subjected to colonial, morphological and biochemical characterization. Pathogenicity test was carried out on the isolates using blood agar to determine their hemolytic activities. Antibiotic resistance status was also investigated against standard antibiotics using the Kirby Bauer disk diffusion method and the multiple antibiotic resistance indices of the isolates were also determined. The plasmid profiles were determined and the resistance genes were identified using molecular means. The bacteria species isolated were Veillonella Spp, Vibrio orientalis and Micrococcus luteus. Veillonella Spp showed Beta hemolysis while Micrococcus luteus showed alpha hemolysis and Vibrio orientalis showed gamma hemolysis. Antibiotic susceptibility test showed all isolates to be antibiotic resistant to varying degrees and multiple antibiotic resistance indices (MARi) depending on their level of resistance. Veillonella. spp. had MARi of 0.3 while M. luteus and V. orietalis had MARi of 0.5. Plasmid profiling showed all isolates to have plasmids with bandwidth of 10,000 bp. Vibrio spp and Veillonella Spp were positive for beta lactamase genes (bla SHV). Only Vibrio was positive for resistance to aminoglycosides (aac(3)-IV). Vibrio orientalis and Micrococcus luteus were found to have the resistance genes for quinolones (qnrA gene) while all the isolates possess genes against the sulphonamides (sul1 gene) used. The result of this work establishes the antibiotic resistance status of bacteria isolated from aquatic habitat and suggests their important role in the spread of antimicrobial resistance AMR, and antibiotic resistance genes (ARGs)

A Dual Beta Blocker and Calcium Channel Blocker Overdose in a Patient with Substance Abuse

Aim: Nearly 25% of hospital admissions have related adverse drug events with 6-7% of these related to beta blockers (BB) or calcium channel blockers (CCB). Introduction: In some cases, patients do not report their noncompliance at home as well as their illicit substance use leading to worsened outcomes. We present a case of BB and CCB toxicity with concomitant substance abuse, its treatment and general outcome. Case: A 49-year-old woman with history of diabetes mellitus, stroke, and hypertension presented with flu-like symptoms including fever, nausea, vomiting, diarrhea and dysuria. She was admitted for treatment of urinary tract infection and possible left lower lobar pneumonia with guideline-directed antibiotic and IV fluids for acute kidney injury. She was elusive about her substance use history and endorsed compliance with her cardiac and other medications. Unfortunately, after administration of her morning medications, she had cardiac decline including hypotension and bradycardia with a subsequent hypoglycemia and left sided deficits leading to further investigation into her medication adherence and substance use history. History evolved that she was noncompliant with her high dose cardiac medications and a urine drug screen was positive for amphetamines, barbiturates, and nonprescription benzodiazepines. Treatment: The patient’s shock was treated with resuscitative fluids, glucagon, and calcium infusion as well as a short round of vasopressor. She was transferred to a higher acuity center for large vessel occlusion at the distal portion of the left vertebral artery as seen on stroke imaging. Her blood and urine cultures ultimately grew extended spectrum beta lactamase bacteria, meanwhile her shock was deemed medication-induced hypovolemic shock with a depressed heart rate response given a normal transthoracic echocardiogram and quick resolution after reversal agent. Despite the severity and progression of the hospitalization the patient elected to leave against medical advice (AMA) prior to neurosurgical evaluation and completion of antibiotics. She has since been seen well in follow up by her outpatient provider. Discussion: Cardiac medications are associated with adverse cardiac effects and should be prescribed carefully in patients with substance abuse. Further elucidation of substance abuse led to improved treatment outcomes in this patient with polysubstance and alcohol abuse.

Prevalence of uropathogen producing extended spectrum beta lactamase (ESBL) at urinary tract infection in chronic kidney disease patients

Background: Urinary tract infection (UTI) is one of the most common bacterial infections encountered by clinicians in developing countries. UTI in patients with Chronic Kidney Disease (CKD) caused by ESBL-producing gram-negative bacteria has been increased in different countries. The common uropathogenic Gram negative bacteria are Escherichia coli and Klebsiella pneumoniae. The aim of this study was to identify prevalence of the common uropathogen producing ESBL E. coli and K. pneumonia among the patients with urinary tract infection (UTI) in chronic kidney disease as well as the susceptibility of antibiotic therapy. Methods: This was a cross-sectional retrospective study to evaluate clinical urine isolates collected from Prof. Dr. I.G.N.G. Ngoerah Hospital within July 2019 until July 2020. Identification and susceptibility tests in urine isolate were performed by Vitex-2 System (Biomerieux) and interpreted with CLSI 2020 standard. Results: From 1291 urine specimens, 210 urine specimens were collected (29.43%) from patients with UTI in CKD during 12 months. Prevalence of uropathogen in chronic kidney disease from urine specimens were E.coli (41.57%), K.pneumoniae (13.15%), and Pseudomonas aeruginosa (7.36%). It was found that ESBL produced E. coli (56.32%) and also K. pneumoniae (54%). Susceptibility antibiotic E. coli and K. pneumonia of UTI patient with CKD with ESBL producers were meropenem (99,10%), amikacin (98,07%), tigecycline (92,30%), gentamicin (80%), and fosfomycin (79,47%). Conclusion: ESBL producers in patient UTI in CKD patients were E. coli 56,32%, K. pneumoniae 54% that were sensitive to meropenem, amikacin, tigecycline gentamicin, fosfomycin.

Antibiotic resistance and consumption before and during the COVID-19 pandemic in Valle del Cauca, Colombia.

To assess changes in antibiotic resistance of eight of the World Health Organization priority bug-drug combinations and consumption of six antibiotics (ceftriaxone, cefepime, piperacillin/tazobactam, meropenem, ciprofloxacin, vancomycin) before (March 2018 to July 2019) and during (March 2020 to July 2021) the COVID-19 pandemic in 31 hospitals in Valle del Cauca, Colombia. This was a before/after study using routinely collected data. For antibiotic consumption, daily defined doses (DDD) per 100 bed-days were compared. There were 23 405 priority bacterial isolates with data on antibiotic resistance. The total number of isolates increased from 9 774 to 13 631 in the periods before and during the pandemic, respectively. While resistance significantly decreased for four selected bug-drug combinations (Klebsiella pneumoniae, extended spectrum beta lactamase [ESBL]-producing, 32% to 24%; K. pneumoniae, carbapenem-resistant, 4% to 2%; Pseudomonas aeruginosa, carbapenem-resistant, 12% to 8%; Acinetobacter baumannii, carbapenem-resistant, 23% to 9%), the level of resistance for Enterococcus faecium to vancomycin significantly increased (42% to 57%). There was no change in resistance for the remaining three combinations (Staphylococcus aureus, methicillin-resistant; Escherichia coli, ESBL-producing; E. coli, carbapenem-resistant). Consumption of all antibiotics increased. However, meropenem consumption decreased in intensive care unit settings (8.2 to 7.1 DDD per 100 bed-days). While the consumption of antibiotics increased, a decrease in antibiotic resistance of four bug-drug combinations was observed during the pandemic. This was possibly due to an increase in community-acquired infections. Increasing resistance of E. faecium to vancomycin must be monitored. The findings of this study are essential to inform stewardship programs in hospital settings of Colombia and similar contexts elsewhere.