Susceptibility Of Bacterial Isolates Research Articles

Determine Biofilm Genes in Pseudomonas Aeruginosa Isolated from Clinical and Environmental Samples

Objective: Isolates of Pseudomonas aeruginosa are an extremely adaptable bacterium that causes opportunistic diseases because of its varied metabolic pathways, genes, virulence factors, and considerable antibiotic resistance. Methods: A total of 293 samples were collected from different places: 193 samples (% 65.87) of human samples and 100 samples (% 34.13) of wastewater samples in the period between 3rd September to 15th November 2023). Bacterial isolates were identified according to microscopic, cultural, and genetic characteristics. Antibiotic susceptibility of bacterial isolates was determined against twelve of the selected antibiotics. The biofilm production was done by using phenotypic ways (Congo red agar and Microtiter plate methods) as well as genotypic ways by detection of biofilm genes (algD, pelf, and pslD genes). Results: Hundred-forty eight bacterial isolates were obtained, and sixty of these isolates were identified as Pseudomonas spp. (40.9%), twenty-six isolates of E. coli (17.9%), seventeen isolates of K. pneumoniae (11.3%), and forty-five isolates were beyond to other types of bacteria (30.1%), and out of sixty isolates of Pseudomonas spp., forty-two were identified as P. aeruginosa isolates. P. aeruginosa isolates revealed various resistance levels to antimicrobial agents gradually, ranging from 83.87% to Trimethoprim-sulfamethoxazole (SXT) to 3.22% to Aztreonam (AZT). Biofilm production by using the Congo red method showed that 27 isolates (64.28%) were positive results, while in the microtiter method, all forty-two isolates were positive (100%), the genetic detection showed that the AlgD gene was recognized in thirty-one isolates (73.8%), followed by Pelf and PslD genes in four isolates each (9.5%). Conclusion: The isolation percentage showed a high occurrence of multi-drug resistance biofilm forming Pseudomonas spp. isolates which could be a critical indicator. Methods of biofilm detection showed that the microtiter plate method has accuracy more than the Congo red method; as well AlgD gene was prevalent compared with both other genes Pelf, and PslD.

Antiviral Drug Candidate Repositioning for Streptococcus suis Infection in Non-Tumorigenic Cell Models.

The increasing prevalence of antimicrobial resistance against zoonotic bacteria, including Streptococcus (S.) suis, highlights the need for new therapeutical strategies, including the repositioning of drugs. In this study, susceptibilities of bacterial isolates were tested toward ten different 3-amidinophenyalanine (Phe(3-Am)) derivatives via determination of minimum inhibitory concentration (MIC) values. Some of these protease inhibitors, like compounds MI-432, MI-471, and MI-476, showed excellent antibacterial effects against S. suis. Their drug interaction potential was investigated using human liver microsomal cytochrome P450 (CYP450) measurements. In our work, non-tumorigenic IPEC-J2 cells and primary porcine hepatocytes were infected with S. suis, and the putative beneficial impact of these inhibitors was investigated on cell viability (Neutral red assay), on interleukin (IL)-6 levels (ELISA technique), and on redox balance (Amplex red method). The antibacterial inhibitors prevented S. suis-induced cell death (except MI-432) and decreased proinflammatory IL-6 levels. It was also found that MI-432 and MI-476 had antioxidant effects in an intestinal cell model upon S. suis infection. Concentration-dependent suppression of CYP3A4 function was found via application of all three inhibitors. In conclusion, our study suggests that the potential antiviral Phe(3-Am) derivatives with 2',4' dichloro-biphenyl moieties can be considered as effective drug candidates against S. suis infection due to their antibacterial effects.

Evaluation of the White Garden Snail (Theba pisana) Mucus Slime for its Efficacy as an Antimicrobial Agent

The escalation of environmental pathogenic microorganisms and disregard of public hygiene practices have resulted in the emergence of various skin infections within communities. Recent investigations have proposed that diverse snail mucus compositions may possess antimicrobial properties. Therefore, it is imperative to conduct further research to elucidate specific antibacterial characteristics inherent in the mucus of White Garden snail (Theba pisana). This study aimed to evaluate antibacterial activity of Theba pisana mucus extract against selected ATCC bacterial strains being Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and MRSA. The assessment involved comparing antimicrobial activity of the mucus extract to various broad-spectrum antibiotics. The susceptibility of bacterial isolates to snail mucus secretions was assessed using agar well diffusion method on Muller Hilton Agar plates. After incubation at 37°C for 24 hours, various concentrations of snail slime including 100%, 75%, 50%, and 25% were applied. The findings reported that E. faecalis exhibited highest activity, with zone of inhibition measuring 24 mm, followed by K. pneumonia and S. aureus (16 mm), MRSA (14 mm), and E. coli (12 mm). However, at concentration of 25%, mucus extract exhibited effect only against E. faecalis. Moreover, the antimicrobial activity of several tested antibiotics demonstrated similarity to that of the mucus extract. Therefore, it revealed that secretions of T. pisana mucus may possess the potential to act as a source of antibacterial agents. This may become as an alternative agent to costly synthetic antibacterial compounds. However, further studies are required to exploit the mucus secretion in addressing the issue of antibiotic resistance.

MICROBIOLOGICAL ANALYSIS AND ANTIBIOTIC SUSCEPTIBILITY FOR SPONTANEOUS BACTERIAL PERITONITIS IN HOSPITALIZED PATIENTS WITH AND WITHOUT HEPATITIS C

In individuals with cirrhosis, spontaneous bacterial peritonitis (SBP) is a common and often fatal bacterial infection with ascites. The Pathological accumulation and collection of fluid in the peritoneal cavity is known as ascites. Ascites can be a challenging diagnosis. Individuals with or without underlying hepatitis C may have varying SBP. Antimicrobials are used to treat SBP, however because MDR organisms are present, initial empirical treatment may not be effective. The aim of the present study was to investigate the causative agents of ascites in SBP and to check antibiotic susceptibility of bacterial isolates. In recent study twenty-five SBP patients were prospectively studied. The ascitic fluid was inoculated in to conventional bacterial broth, agar culture and blood culture media. Among all tested bacterial species E. coli was the most isolated bacteria present in 23 samples out of 25. The presence of E.coli was 92% followed by Shigella spp (72%), Salmonella spp (68%), Enterobacter spp (60%), K. aerogenes (52%), S. aureus (32%), S. epidermidis (30%), A. israelii (28%), P. aeruginosa (24%), C. freundii (20%) and S. marcescens (08%). A total of six antibiotics were used to check antibiotic susceptibility pattern of bacterial isolates through disc diffusion method. The used antibiotics were Meropenem, Moxifloxacin, Ciprofloxacin, Ceftriaxone, Cefixime and vancomycin. Our results showed that most of isolated bacterial species were MDR (Multi drug resistant) except E.coli and A.israelii that were sensitive to used antibiotics. Meropenem was the most effective antibiotic against all tested bacterial species. Meropenem showed maximum zone of inhibition against S.auresus (34mm), followed by A. israelii (33mm), P.aeruginosa (32mm), C.freundii (31mm), S.marcescens (28mm), S.epidermidis (27mm), E.coli (26mm), Salmonella spp (23mm), K.aerogenes (21mm), Enterobacter spp (21mm) and Shigella spp (15mm). It is concluded that gram-positive and gram-negative multi-drug resistant bacteria were the most common cause of SBP in patients with and without hepatitis C and meropenem was the most effective antibiotic against all tested isolate

COMPARISON OF SUSCEPTIBILITY BETWEEN CONVENTIONAL FIRST LINE ANTIBIOTIC CO-TRIMOXAZOLE AND NEWER ANTIBIOTICS IN RECURRENT UNCOMPLICATED URINARY TRACT INFECTIONS

Objective: The current study aims to evaluate the susceptibility pattern of relatively older antibiotic Co-trimoxazole and its comparison with Levofloxacin, Nitrofurantoin, and Fosfomycin. Material and Methods: This cross-sectional study was conducted in Mardan Medical Complex, Mardan, and the Postgraduate Medical Education Department of Khyber Girls Medical College, Peshawar from April 2022 to September 2022. Both male and female patients, above the age of 15 years with recurrent uncomplicated urinary tract infections were included in the study. The samples were inoculated onto CLED (Cystine-Lactose-Electrolyte-Deficient) Agar, a differential culture medium. The grown bacteria were identified, using Gram staining and BIOMÉRIEUX® API® 10S kits. Minimum inhibitory concentrations (MIC) were determined by the Agar dilution method; as per standard protocol. The results were compared among Co-trimoxazole, Levofloxacin, Nitrofurantoin, and Fosfomycin using statistical tests. Results: A total of 680 samples were received, of which 158 samples were culture-positive. The gender distribution of females and males was 63.3% and 36.7%, respectively. A predominant proportion of the patient cohort manifested within the age range of 21-40 years, with the subsequent highest representation observed in the 41-60 year age group. The isolated organisms were E. coli (74.1%), Klebsiella (10.8%), Pseudomonas (5.1%), Enterococci (6.3%), Proteus species (2.5%), and Citrobacter (1.3%). Based on MIC analysis, 77.2% of isolates were found to be sensitive to Co-trimoxazole, 52.5% to Levofloxacin, 86.7% to Nitrofurantoin, and 90.5% to Fosfomycin. When comparing antibiotics, Co-trimoxazole displayed significantly higher effectiveness against the isolates compared to Levofloxacin (p-value 0.004). However, in comparison to Nitrofurantoin and Fosfomycin, Co-trimoxazole exhibited lower effectiveness, with respective p-values of 0.000 and 0.007. Conclusion: The study revealed that the susceptibility of bacterial isolates to Co-trimoxazole is significantly higher than that to Levofloxacin but lower than that to Nitrofurantoin and Fosfomycin. KEYWORDS Urinary Tract Infections, Drug Resistance, Culture And Sensitivity, Minimum Inhibitory Concentration

Uncomplicated bacterial uroinfections: aetiological structure and antimicrobial susceptibility of bacterial isolates

Introduction and objective: The increasing resistance to antimicrobials also affects uropathogens, the causative agents of uncomplicated urinary tract infections (UTIs), and is a frequent cause of failure of empirical antimicrobial therapy. The purpose of the study is to analyze the aetiological structure and susceptibility to antimicrobials of the most common bacterial causative agents of UTIs. Material and Methods: A prospective analysis of microbial findings was performed in patients with clinical evidence of UTIs. A total of 9,426 urine cultures were examined (female: 5,157; male: 4,269). Results: Microbial growth was found in 29.7% of female urine cultures and in 19.1% of male urine cultures. A total of 2,364 microbial isolates were identified. The aetiological structure of the isolates in both groups was similar – female: E. coli 51.3 %, Enterococcus spp. 18.9%, K. pneumoniae 13.1%, P.mirabilis 4.6%, Candida spp. 1.2%; male: E. Coli 23.5 %, Enterococcus spp.27.4%, K. pneumoniae 19.5%, P. mirabilis 6.4%, Candida spp. 2.8%. A high frequency of resistance was found to cephalosporins III generation (ceftriaxone: K.pneumoniae 31.7 %, P. mirabilis 15.2 %), fluoroquinolones (ciprofloxacin: E.faecium 74.0 %, K.pneumoniae 34.4 %, E. faecalis 27.9 %, E. coli 27.0 %, P. Mirabilis 24.0 %) and trimethoprim/sufamethoxazole (K. pneumoniae 33.9 %, P. mirabilis 33.6 %, E. coli 25.3 %,). Resistance to fosfomycin and nitrofurantoin was low. ESBL-producers were more frequent among K. pneumoniae (31.7 %) compared to E. coli (9.3 %). Resistance to carbapenems was found only in K. pneumoniae (14.9 %). Conclusion: The study showed that the main uropathogens are often resistant to the antimicrobials recommended for empiric antimicrobial therapy in UTIs, necessitating a revision of local recommendations.

Microbial Profiles and Patterns of Antibiotic Susceptibility of Bacterial Isolates from Edible Sprouts of Flaxseeds (Linum usitatissimum L.)

Flaxseed (Linum usitatissimum L.) is a plant-based food that provides healthy fat, antioxidants, and fiber. It is referred to as a "functional food" by certain individuals, which suggests that someone can eat it to improve their health. A study on sprouted flaxseed produced in the laboratory revealed the presence of microbial flora, namely bacteria and molds, which is of concern for the health-conscious public. A total of 39 flaxseed sprout samples were cultured to evaluate microbial profiles and to determine antibiotic susceptibility patterns by the Kirby-Bauer Disc diffusion method. Bacterial isolates were tested only against 11 antibiotics, viz. Amoxycillin, Gentamicin, Ciprofloxacin, Colistin, Meropenem, Cefixime, Azithromycin, Mecillinam, Cotrimoxazole, Cefaclor, and Moxifloxacin. 39 (100%) of the 39 flaxseed sprout samples cultured tested positive for bacteria and molds. Among them, positive cultures, single bacterial growth, multiple bacterial growth, and molds were 33 (84.6%), 6 (15.4%) and 39 (100%) respectively. The most predominant organisms were Serratia marcescens, comprising 15(33.3%), followed by Citrobacter freundii 10(22.2%), Pseudomonas aeruginosa 9(20.0%), and other bacterial isolates were Enterobacter cloacae complex, Morganella morganii, and Aeromonas hydrophila, whose frequencies were 6(13.3%), 4(8.9%) and 1(2.2%) respectively. All of the bacterial isolates were 100% sensitive to Ciprofloxacin, Gentamicin, Mecillinam, and Moxifloxacin, and only two antibiotics were shown 100% resistance in Amoxicillin and Cefixime. Based on the results, it is suggested that consumers should pay attention to producing with hygienic techniques and cooking properly to avoid foodborne diseases from flaxseed sprouts
 J. of Sci. and Tech. Res. 4(1): 155-164, 2022

An In Vitro Susceptibility Study of Cefotaxime-Sulbactam on Clinical Bacterial Isolates From Various Regions in India: A Comparison With Ceftriaxone-Sulbactam.

Background and objective Combining sulbactam with cefotaxime/ceftriaxone augments its antimicrobial activity against β-lactamase-producing bacteria. They are widely used as empirical treatment for many clinical infections. However, there is a scarcity of data on the susceptibility of various organisms to these antibiotics in the Indian region. In light of this, the present in vitro study evaluated the susceptibility of bacterial isolates to cefotaxime-sulbactam and compared it withceftriaxone-sulbactam. Methodology Clinical samples with positive bacterial cultures from various laboratories in India were subjected to antibiotic sensitivity testing using in vitro E-test strips and disk diffusion methods to determine the minimum inhibitory concentration (MIC) and zone of inhibition (ZOI), respectively.MIC50 and MIC90 values were determined along with the measurement of the ZOI for the effectiveness of antibiotics. Interpretations of MIC and ZOI values were made as per the criteria set by the Clinical and Laboratory Standards Institute (CLSI) guidelines to estimate the proportion of sensitive organisms. Results Among 400 clinical isolates evaluated, Escherichia coli (E. coli)(47.75%) was the most common organism isolated followed by Klebsiella (26%), Salmonella (7.75%), Proteus (3.8%), and Acinetobacter (2.8%). The mean ZOI was found significantly higher for E. coli, Klebsiella, and Salmonella in the cefotaxime-sulbactam group than in the ceftriaxone-sulbactam group. MIC50 values for E. coliand Klebsiellawere 0.25and 0.19 µg/ml, respectively in the cefotaxime-sulbactam group as compared to 0.38and 0.25 µg/ml, respectively for ceftriaxone-sulbactam. The proportion of sensitive isolates was also higher in the cefotaxime-sulbactam group forE. coli, Klebsiella, and Salmonella. Conclusions The in vitro effect of cefotaxime-sulbactamon organisms is similar to that of ceftriaxone-sulbactam in terms of MIC, ZOI, and proportion of sensitivity based on our study involving clinical isolates fromvarious parts of India. Cefotaxime-sulbactam may be preferred in the empirical management of various clinical infections.

EFFICIENCY OF SOME CHEAP MATERIALS IN REMOVING SOME HEAVY METALS

Investigation tests showed the susceptibility of bacterial isolates (Leuconostoc mesenteroids spp.mesenteroids ATCC829, Leuconostoc mesenteroids, Leuconostoc mesenteroids ssp.dextranicum, Lactobacillus casei, Lactobacillus rhamosus GG, Lactobacillus plantarum, Leuconostoc mesenteroids) to biofilm formation using titration microplates. In removing metals, it was chosen in conducting subsequent experiments, and the live mass of the two isolates Lactobacillus casei and Leuconostoc mesenteroids showed better retention of lead, cadmium, and cobalt than the biofilm of which it is composed. The results of restriction using multiple materials (sodium alginate, white sand, glass sand, sugar cane residues, activated carbon, Orbeez balls, silica gel, sponge, sunflower seed husks, date kernels, and coconut residues) showed positive results in the removal process. The restriction of adsorption-efficient bacterial cells using Orbeez balls and silica gel was shown to be the best substrate compared to other restrictive substrates

Antimicrobial susceptibility of bacterial isolates from clinical specimens in four Pacific Island countries, 2017–2021

There are limited antimicrobial resistance (AMR) surveillance data from low- and middle-income countries, especially from the Pacific Islands region. AMR surveillance data is essential to inform strategies for AMR pathogen control. We performed a retrospective analysis of antimicrobial susceptibility results from the national microbiology laboratories of four Pacific Island countries - the Cook Islands, Kiribati, Samoa and Tonga - between 2017 and 2021. We focused on four bacteria that have been identified as 'Priority Pathogens' by the World Health Organization: Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. Following deduplication, a total of 20,902 bacterial isolates was included in the analysis. The most common organism was E.coli (n=8455) followed by S.aureus (n=7830), K.pneumoniae (n=2689) and P.aeruginosa (n=1928). The prevalence of methicillin resistance among S.aureus isolates varied between countries, ranging from 8% to 26% in the Cook Islands and Kiribati, to 43% in both Samoa and Tonga. Ceftriaxone susceptibility remained high to moderate among E.coli (87%-94%) and K.pneumoniae (72%-90%), whereas amoxicillin+clavulanate susceptibility was low against these two organisms (50%-54% and 43%-61%, respectively). High susceptibility was observed for all anti-pseudomonal agents (83%-99%). Despite challenges, these Pacific Island laboratories were able to conduct AMR surveillance. These data provide valuable contemporary estimates of AMR prevalence, which will inform local antibiotic formularies, treatment guidelines, and national priorities for AMR policy. Supported by the National Health and Medical Research Council.

Susceptibility of Bacterial Isolates From Wound Swabs in Bangladesh: Laboratory – Based Surveillance Study

Background: Wound infection is one of the major health problems that occur frequently. Infections of the wound result from entry of the organisms through breached skin. It plays an important role in the development of chronicity, delaying wound healing. It is evident that wound infection is a challenging situation for the physicians. Multiple bacteria can cause wound infection. Both broad spectrum and narrow spectrum antibiotics are available for the treatment. It is ideal to give proper antibiotic after culture and sensitivity of the wound swab. Improper and irrational use of antibiotics can lead to drug resistance. Objective: To isolate and identify the bacteria causing wound infection and to determine the antimicrobial susceptibility pattern. Materials and method: This descriptive cross-sectional study was carried out at the Microbiology Laboratory, Popular Diagnostic Centre Ltd, Dhanmondi, Dhaka, Bangladesh, from November 2019 to January 2020. A total of 62 wound swabs were collected and analyzed for culture and antibiotic sensitivity. Results: From total wound swab samples, 49 (79%) were culture positive. The most common isolated pathogen was Acinetobacter spp. 20 (40.81%). Other isolates were Staphylococcus aureus 10 (20.41%); Escherichia coli 07 (14.29%); Pseudomonas aeruginosa 07 (14.29%); Klebsiella 04 (08.16%) and Proteus 01 (02.04%). Among the Gram negative isolates, Acinetobacter spp showed 100% sensitivity to colistin, 85% to imipenem, 30% to amikacin and 25% to piperacillin + tazobactum. E. coli showed 100% sensitivity to imipenem and colistin and least sensitivity to third generation cephalosporin. Pseudomonas showed 100% sensitivity to piperacillin+tazobactum, 85.71% to imipenem and aztreonam. Among the Gram positive isolates, Staphylococcus aureus showed 100% sensitivity to linezolid, vancomycin, imipenem, cloxacillin, amikacin and least sensitivity to azithromycin. Conclusion: Acinetobacter spp. was the most frequently isolated pathogen from wound swab and the antibiotic sensitivity pattern of various isolates will help the clinician in appropriate selection of antibiotics against wound infection. J Dhaka Med Coll. 2021; 30(2) : 180-188

Informing antimicrobial stewardship with explainable AI.

The accuracy and flexibility of artificial intelligence (AI) systems often comes at the cost of a decreased ability to offer an intuitive explanation of their predictions. This hinders trust and discourage adoption of AI in healthcare, exacerbated by concerns over liabilities and risks to patients' health in case of misdiagnosis. Providing an explanation for a model's prediction is possible due to recent advances in the field of interpretable machine learning. We considered a data set of hospital admissions linked to records of antibiotic prescriptions and susceptibilities of bacterial isolates. An appropriately trained gradient boosted decision tree algorithm, supplemented by a Shapley explanation model, predicts the likely antimicrobial drug resistance, with the odds of resistance informed by characteristics of the patient, admission data, and historical drug treatments and culture test results. Applying this AI-based system, we found that it substantially reduces the risk of mismatched treatment compared with the observed prescriptions. The Shapley values provide an intuitive association between observations/data and outcomes; the associations identified are broadly consistent with expectations based on prior knowledge from health specialists. The results, and the ability to attribute confidence and explanations, support the wider adoption of AI in healthcare.

Microbiological Profile of Blood Stream Infections in a Tertiary Care Hospital, J&K. India

Background: Worldwide, bacteremia is one of the serious infections that cause significant morbidity and mortality in hospitalized patients. A wide range of microorganisms have been implicated in the etiology of BSIs. Early detection of these pathogens along with the determination of antimicrobial susceptibility patterns have been shown to improve treatment outcomes. The present study aimed to determine the microbiological profile of BSIs in a major tertiary care hospital in North India.
 Materials and Methods: This is a cross-sectional study conducted in the Department of Microbiology Government Medical College Srinagar. Blood samples submitted in brain heart infusion (BHI) broth for culture and sensitivity over a period of one year from September 2021 to 30th August 2022 were included in the study and processed per standard laboratory protocol techniques for isolating and identifying pathogens causing BSI. The antimicrobial susceptibility of bacterial isolates was determined by the disc diffusion method as per Clinical and Laboratory Standards Institute (CLSI) guidelines.
 Results: Out of 4260 blood samples, 1204(28.26%) isolates were obtained. Of these isolates, 575(47.75%) were Gram‑positive bacteria and 468 (38.87%) were Gram‑negative bacteria. Candida species were isolated from 161(13.7%) positive samples and 32 showed contamination. The most commonly identified organism was Coagulase‑negative Staphylococcus (CoNS) (22.59%) followed by Staphylococcus aureus(17.69%), Acinetobacter spp. (13.70%), Klebsiella spp (10.13%) and Escherichia coli (6.47%). Among the gram-positive bacteria, maximum resistance was seen with methicillin and azithromycin. No resistance was seen with vancomycin and linezolid. Most of the gram-negative bacilli were multidrug-resistant. Maximum resistance was seen with ampicillin(91.7%), amoxiclav (86.5%), ceftriaxone (88.5%), and gentamicin (60.9%). Higher prevalence of resistance was observed in Gram‑negative bacteria when compared with Gram‑positive bacteria.
 Conclusion: Hence, empirical treatment of BSIs should be based on the current knowledge of bacterial resistance profiles as provided by microbiology laboratory reports. The results of this study warrant continuous monitoring of antimicrobial patterns for the clinicians along with judicious antibiotic policy to mandate antimicrobial sensitivity testing against the BSIs in the hospital setup so that appropriate therapeutic measures should be taken at the earliest.

Chlorhexidine and octenidine susceptibility of bacterial isolates from clinical samples in a three-armed cluster randomised decolonisation trial.

Routine use of chlorhexidine or octenidine for antiseptic bathing may have unintended consequences. Our analysis aimed to assess the phenotypic susceptibility of bacterial isolates from clinical samples to chlorhexidine and octenidine collected from intensive care units (ICU) that routinely used 2% chlorhexidine-impregnated wash cloths or 0.08% octenidine wash mitts (intervention) or water and soap (control) for daily patient care. This study was conducted within the context of a three armed cluster-randomised controlled decolonisation trial (Registration number DRKS00010475, registration date August 18, 2016). Bacterial isolates were collected prior to and at the end of a 12-month-intervention period from patients with ≥ 3 days length of stay at an ICU assigned to one of two intervention groups or the control group. Phenotypic susceptibility to chlorhexidine and octenidine was assessed by an accredited contract research laboratory determining minimal inhibitory concentrations (MIC) as percentage of extraction solutions used. MIC were reported as estimated concentrations in μg/ml derived from the chlorhexidine and octenidine extraction solutions. Statistical analyses including generalized estimating equation models were applied. In total, 790 ICU-attributable bacterial isolates from clinical samples (e.g. blood, urine, tracheal aspirate) were eligible for all analyses. Pathogens included were Staphylococcus aureus (n = 155), coagulase-negative staphylococci (CoNS, n = 122), Escherichia coli (n = 227), Klebsiella spp. (n = 150) and Pseudomonas aeruginosa (n = 136). For all species, chlorhexidine and octenidine MIC did not increase from baseline to intervention period in the antiseptic bathing groups. For proportions of bacterial isolates with elevated chlorhexidine / octenidine MIC (≥ species-specific chlorhexidine / octenidine MIC50), adjusted incidence rate ratios (aIRR) showed no differences between the intervention groups and the control group (intervention period). We found no evidence for reduced phenotypic susceptibilities of bacterial isolates from clinical samples to chlorhexidine or octenidine in ICUs 12 months after implementation of routine antiseptic bathing with the respective substances.

Molecular Analysis of Mineral and Tap Water to Detect Microbial Contaminants in Jeddah Province

Since ancient times, drinking water quality has played a significant role in human diseases and infections. In Jeddah, the drinking water choices are mostly bottled water or water available in plastic containers after processing at private water treatment stations. A total of 30 drinking water samples were obtained from many sources in Jeddah, KSA: mineral water (13 samples) and tap water (17 samples). Microbiological tests were used, including a negative and positive Gram stain test for these colonies and determining the type of bacteria. To identify bacterially, Polymerase chain reaction (PCR) techniques were used in this study. Following that, the susceptibility of bacterial isolates to commonly used antibiotics is tested. The results showed that the microbial cultures were of different isolates. We confirmed that these results by microbiological examination and molecular identification found a difference between the study samples. Fifty-three bacteria were isolated from 30 samples, 34 (64.15%) tap water samples, and 19(35.85%) mineral water samples were found to be contaminated with bacteria. The highest bacterial contamination, 86.6% of all samples, was found in the tap water in 50% of samples, while 36.6% was found in mineral water. Microbiological tests and molecular diagnostic methods make it simpler and faster to shorten the detection time of drinking water contamination. The results of the current study suggest that mineral water is a better source of potable water in Jeddah than tap water. However, there may be pollution in the water at the reservoirs rather than at the water sources

Antibiotics Resistance Pattern of Food-Borne Bacteria Isolated from Ice Cream in Bangladesh: A Multidisciplinary Study

Ice cream is one of the most popular food items consumed during the summer season in Bangladesh by all ages but mostly school-going students. Due to the ingredients and handling process of ice cream, it acts as a good shelter for pathogenic and nonpathogenic microorganisms. Therefore, we aimed to investigate the microbial count, prevalence, isolate and characterize multidrug-resistant bacteria in ice cream samples collected from nearby shops of schools in Tangail district, Bangladesh. Ice-cream consumer and nonconsumer students were selected by providing questionnaires. Total viable count (TVC) and total coliform count (TCC) were determined by pour plate methods, where conventional methods were performed for bacterial identification. The Kirby-Bauer disk diffusion method was used to determine the antimicrobial susceptibility of bacterial isolates. Kado and Liu method, with some modifications, was used to extract plasmid from the isolated bacteria and visualized through gel electrophoresis. The demographic characteristics showed that the degree of symptoms regarding microorganisms mediated disorders and rate of antibiotics intake in ice cream consumers were significantly higher than the nonconsumers. The range of TVC and TCC in the ice cream samples was found 0–9.9 × 109 CFU/ml and 0–900 CFU/ml, respectively. Interestingly, 93.75% of the total ice cream samples also showed fungal positive. A total of 12 different bacterial species were identified, including Proteus spp, E. coli, V.cholera, Pseudomonas spp, Shigella spp, Klebsiella spp, Aeromonas spp, V. Parahemolyticus. Salmonella paratyphi, Citrobacterspp, Plesiomonasspp, and Staphylococcus aureus. The antimicrobial susceptibility assay showed the multiple resistance frequency of these isolates to different antimicrobial drugs. All individual isolates were screened for plasmid DNA, and we found that seven strains harbored a single or more than two plasmids sized approximately between 1.9 and 140 MDa, indicating a possible connection between resistance phenotype pattern and genotype.

Antimicrobial Activity of Mixed Lactobacilli in Combination with Antibiotics Against Bacterial Pathogens Causing Chronic Suppurative Otitis Media

This study aimed to determine the antimicrobial activity of CFS of the selected lactobacilli, alone and in combination with antibiotics against pathogenic bacteria isolated from patients with chronic otitis media. In addition, we investigated the ability of tested lactobacilli (a mixed of Lactobacillus acidophilus and Lactobacillus plantarum) to co-aggregate with the isolated pathogens.. One hundred sixty-two ear swab were collected from patients had otitis media with chronic discharging . These patients attended to the ENT department in the medical consultation clinic at Baquba Teaching Hospital and to private doctors’ clinics. Ear swabs were taken from both sexes, male and female starting from September to the end of December, 2021. The isolates were identified based on their morphological features of the colonies and some biochemical tests. The final identification and antibiotics susceptibility of bacterial isolates was confirmed by VITEK2 compact system. The results showed that 113 samples were positive for bacterial culture, while 30 samples were negative growth, mostly identified as fungi. A total of 113 bacterial species isolated from 162 otitis media samples were: Pseudomonas aeruginosa, Staphylococcus aureus, Proteus mirabilis, and Klebsiella pneumonia. The highest predominant bacterial agents were P. aeruginosa 52 (32%), S. aureus 40 (24.7%), followed by P. mirabilis 14 (8.64%), and K. pneumoniae 7 (4.32%). The obtained results showed that the highest rate of infection was identified in male rather than in female. In antibiotic susceptibility testing, 16 types of antibiotics was evaluated against four bacterial species using VITEK2 system. In addition, disk diffusion assay was utilized in order to perform antimicrobial combinations. The susceptibility of bacterial species were various among these antibiotics. It was noticed that ciprofloxacin was the most effective antibiotic against all the isolated bacteria with the lowest MIC in comparison to the other antibiotics. We evaluated the effect of ciprofloxacin alone and in combination with the cell-free supernatants (CFSs) of mixed lactobacilli against the four isolates using disk diffusion assay. Our results showed that the CFS produced by the mixed lactobacilli have a significant inhibitory effect on the growth of bacterial pathogens. Moreover, CFS in combination with ciprofloxacin was more effective against pathogenic isolates than using ciprofloxacin alone. Co-aggregation of the selected mixed lactobacilli strains with the four bacterial isolates were evaluated in order to investigate the competitive activity of the tested lactobacilli with the isolated pathogens. Keywords: Otitis media, Mixed lactobacilli, Cell Free Supernatant, Co-aggregation, Antimicrobial combinations

Urinary Tract Infection and Antimicrobial Susceptibility of Bacterial Isolates in Saint Joseph Kitgum Hospital, Kitgum, Uganda

A cross-sectional study of microorganisms isolated from mid-stream urine samples obtained from 139 patients with suspected urinary tract infection (UTI) who presented leukocyturia was conducted from April to June 2019 at Saint Joseph Kitgum Hospital (Uganda). All microorganisms were identified by MALDI-TOF mass spectrometry in a laboratory in Spain. Antimicrobial susceptibility was determined on site using the disc diffusion method (Kirby–Bauer test) and these results were subsequently compared with those obtained in Spain using the Becton Dickinson Phoenix M50 device. The overall prevalence of UTI with bacterial growth was 64.0% (n = 89) (95% CI, 56.1–72.0), and 11 presented mixed infection. As a result, 100 microorganisms were isolated. The most common uropathogens were Enterococcus spp. (57%) and Escherichia coli (28%). Nitrofurantoin was the most effective drug (81.7% in Gram-positive and 87.3% in Gram-negative bacteria), followed by imipenem (94.2% and 74.5%, respectively). The highest resistance rates were observed for amoxicillin and ciprofloxacin (66.2% and 44.6%, respectively). Given the increasing trend toward antibiotic resistance, there is a need for bacteriological cultures and continuous surveillance of uropathogen antibiotic susceptibility. Use of amoxicillin and ciprofloxacin as empirical treatments for UTIs should be discontinued in Uganda. The findings of this study may be useful for clinicians, as they may improve empirical treatment.

Analysis in Results of Microbiologic Exam Related to Donor Corneas

Purpose: This study analyzed the microorganisms identified in donor corneas and their clinical significance.Methods: The medical records of 94 patients (114 eyes) who underwent keratoplasty and microbiological tests of the donor corneas from October 2008 to December 2020 at our hospital were reviewed retrospectively. During keratoplasty, we conducted microbiological tests of the corneoscleral rim of the donor cornea and preserving solution Optisol™-GS (Bausch & Lomb, Rochester, NY, USA), and examined the antibiotic susceptibility of bacterial isolates from the cultures.Results: Some isolates of domestic donor corneas revealed bacteria, but none of the imported corneas did. Gram-negative bacilli were detected from the corneoscleral rim in three eyes (2.6%): two cases of Acinetobacter baumannii/haemolyticus and one case of Pseudomonas aeruginosa. In one case (0.9%), Acinetobacter baumannii/haemolyticus was identified from preserving solution, with no bacteria found in the corneoscleral rim. Antibiotic susceptibility tests showed multi-drug resistance, except to colistin. In all cases where bacteria were detected, there was no keratitis or endophthalmitis after corneal transplantation.Conclusions: Although rare, bacteria can be identified from donor corneas or the preserving solution used in corneal transplantation. Therefore, caution is needed in all processes dealing with donor corneas. Empirical antibiotics that have sufficient antimicrobial activity to suppress multi-drug resistant bacteria should be selected in corneal transplantation.

DIAGNOSIS OF E.COLI ISOLATED FROM ARTHRITIS IN CHICKENS BY VITEK AND MOLECULAR METHODS

This study was aimed to identify and confirm of E.coli isolated from arthritis in chickens. Samples for the isolation of the bacteria were taken from broiler chickens with arthritis symptoms (swelling hock joint), and then examined by culturing, VITEK test as well as molecular assay. Antimicrobial susceptibility of bacterial isolate was done. The results showed pink colour colonies on MacConkey agar. The VITEK system was used to identify those colonies. The results revealed that the isolate gave 97 % parallel to those features of E. coli equally recognized by the standard gram negative card. The isolate was found to be sensitive to ticarcillin/clavulanic acid and many others antibiotics and resistance to ticarcillin as well as for other types of antibiotics .The results of the 16Sr RNA gene revealed that E.coli primers of the16S rRNA gene had successfully targeted the respective gene and have shown the single bands of the16S RNA gene at 1500 bp. Sequencing of this gene was performed for the isolate. The results of nucleotide sequencing were submitted in Gene bank database and have accession number: ID: MT012194.1. The phylogenetic analysis Of the isolate was 100% similar to USA:CP048605, Pakistan:GU594300, VitNamHatey: AP022650 and China:MN208222, however, it was 99% similar to India:LCO58573.