Search results for: multi antibiotic resistance

Authors: Stelvin Sebastian, Andriya Annie Tom, Joyalanna Babu, Merin Joshy

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Introduction: The worldwide increase in the use of antibiotics in poultry and livestock industry to treat and prevent bacterial diseases and as growth promoters in feeds has led to the problem of development of antibiotic resistance both in animals and human population. Aim: To study the pattern of antibiotic use and prevalence of multidrug-resistant Escherichia coli in poultry chickens in selected farms in Muvattupuzha and to compare the spread of multidrug-resistant bacteria from poultry environment to UTI patients. Methodology: Two farms from each of 6 localities in Muvattupuzha were selected. A questionnaire on the pattern of antibiotic use and various farming practices were surveyed from farms. From each farm, 60samples of fresh fecal matter, litter from inside, litter from the outside shed, agricultural soil and control soil were collected, and antimicrobial susceptibility testing of E. coli was done. Antibiogram of UTI patients was collected from the secondary care hospital included in the study, and those were compared with resistance patterns of poultry samples. Results: From survey response antibiotics such as ofloxacin, enrofloxacin, levofloxacin, ciprofloxacin, colistin, ceftriaxone, neomycin, cephalexin, and oxytetracycline were used for treatment and prevention of infections in poultry. 31of 48 samples (51.66%) showed E. coli growth. 7 of 15 antibiotics (46.6%) showed resistance. Ampicillin, amoxicillin, meropenem, tetracycline showed 100% resistance to all samples. Statistical analysis confirmed similar resistance pattern in the poultry environment and UTI patients for antibiotics such as ampicillin, amoxicillin, amikacin, and ofloxacin. Conclusion: E. coli were resistant not only to extended-spectrum beta-lactams but also to carbapenems, which may be disseminated to the environment where litter was used as manure. This may due to irrational use of antibiotics in chicken or from their use in poultry feed as growth promoters. The study concludes the presence of multidrug-resistant E.coli in poultry and its spread to environment and humans, which may cause potentially serious implications for human health.

Keywords: multidrug resistance, escherichia coli, urinary tract infection, poultry

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Authors: Solomon Gebremariam, Mulugeta Naizigi, Aregawi Haileselassie

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Background: Knowledge of the pattern of bacterial isolates and their antimicrobial susceptibility is crucial for guiding empirical treatment and infection prevention and control measures. Objective: The aim of this study was to analyze the pattern of bacterial isolates and their susceptibility patterns from various specimens. Methods: Retrospectively, a total of 1067 microbiological culture results that were isolated, characterized, and identified by standard microbiological methods and whose antibiotic susceptibility was determined using CLSI guidelines between 2017 and 2019 were retrieved and analyzed. Data were entered and analyzed using the Stata release 10.1 statistical package. Result: The positivity rate of culture was 26.04% (419/1609). The most common bacteria isolated were S. aureus 23.8% (94), E. coli 15.1% (60), Klebsiella pneumonia 14.1% (56), Pseudomonas aeruginosa 8.5% (34), and CONS 7.3% (29). S. aureus and CONS showed a high (58.1% - 96.2%) rate of resistance to most antibiotics tested. They were less resistant to Vancomycin which is 18.6% (13/70) and 11.8% (2/17), respectively. Similarly, the resistance of E. coli, Klebsella pneumonia, and Pseudomonas aeruginosa was high (69.4% - 100%) to most antibiotics. They were less resistant to Ciprofloxacilin, which is 41.1% (23/56), 19.2% (10/52), and 16.1% (5/31), respectively. Conclusion: This study has shown that there is a high rate of antibiotic resistance among bacterial isolates in this hospital. A combination of Vancomycin and Ciprofloxacin should be considered in the choice of antibiotics for empirical treatment of suspected infections due to S. aureus, CONS, E. coli, Klebsiella pneumonia, Pseudomonas such as in infections within hospital setup.

Keywords: antimicrobial, resistance, bacteria, hospital

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Authors: M. Ottenbrite, G. Yilmaz, J. Devenish, M. Kang, H. Dan, M. Lin, C. Lau, C. Carrillo, K. Bessonov, J. Nash, E. Topp, J. Guan

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Consumption of food contaminated by antibiotic-resistant (AR) bacteria may lead to the transmission of AR genes in the gut microbiota and cause AR bacterial infection, a significant public health concern. However, information is limited on if and how background microbes from the food matrix (food microbes) may influence resistance transmission. Thus, we assessed the colonization of a β-lactam resistant Salmonella Heidelberg strain (donor) and a β-lactam susceptible S. Typhimurium strain (recipient) and the transfer of the resistance genes in the mouse gut in the presence or absence of food microbes that were derived from washing freshly-harvested carrots. Mice were pre-treated with streptomycin and then inoculated with both donor and recipient bacteria or recipient only. Fecal shedding of the donor, recipient, and transconjugant bacteria was enumerated using selective culture techniques. Transfer of AR genes was confirmed by whole genome sequencing. Gut microbial composition was determined by 16s rRNA amplicon sequencing. Significantly lower numbers of donors and recipients were shed from mice that were inoculated with food microbes compared to those without food microbe inoculation. S. Typhimurium transconjugants were only recovered from mice without inoculation of food microbes. A significantly higher survival rate was in mice with vs. without inoculation of food microbes. The results suggest that the food microbes may compete with both the donor and recipient Salmonella, limit their growth and reduce transmission of the β-lactam resistance gene in the mouse gut.

Keywords: antibiotic resistance, gene transfer, gut microbiota, Salmonella infection

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Authors: S. K. Al-Musharafi, I. Y. Mahmoud, S. N. Al-Bahry

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At Ras Al-Hadd Reserve, Eggs from green turtles and Chelonia mydas were randomly collected immediately after Oviposition. Eggshells taken from fresh eggs and sand collected from the body chamber were analyzed for eight heavy metals (Al, Br, Cd, Co, Cu, Fe, S, and Zn) using inductively coupled plasma mass spectrometry (ICP). Heavy metal concentrations varied significantly (P<0.05) between nest sand and eggshells. Zn values were significantly higher than the other heavy metals. A total of 60 heterotrophic bacteria belong to eight genera were isolated from fresh egg contents (albumen and yolk). Resistance of the isolates to Amikacin, ampicillin, chloramphenicol, gentamycine, minocylin, nalidixicacid, neomycin, streptomycin, tetracycline, tobramycin, and Trimethoprim was tested. More than 40 % of the isolates were multiple resistant to 2-7 antibiotics. Most of the resistant strains were also resistant to Zn. The value of these findings may indicate that the origin of pollution is of human contaminated effluents.

Keywords: antibiotic resistance, bacteria, environment, heavy metals, sea turtles

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Authors: Natesan Balasubramanian, Palpandi Pounpandi, Venkatraman Thamil Priya, Vellasamy Shanmugaiah, Karubbiah Balakrishnan, Mandayam Anandam Thirunarayan

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Streptococcus agalactiae is commonly known as Group B Streptococcus (GBS) and it is the most common cause of life-threatening bacterial infection. GBS first considered as a veterinary pathogen causing mastitis in cattle later becomes a human pathogen for severe neonatal infections. In this present study, a total of 20 new clinical isolates of S. agalactiae were collected from male (6) and female patient (14) with different age group. The isolates were from Urinary tract infection (UTI), blood, pus and eye ulcer. All the 20 S. agalactiae isolates has clear hemolysis properties on blood agar medium and were identified by serogrouping and MALTI-TOF-MS analysis. Antibiotic susceptibility/resistance test was performed for 20 S. agalactiae isolates, further phenotypic resistance pattern was observed for tetracycline, vancomycin, ampicillin and penicillin. Genotypically we found two antibiotic resistance genes such as Betalactem antibiotic resistance gene (Tem) (70%) and tetracycline resistance gene Tet(O) 15% in our isolates. Six virulence factors encoding genes were performed by PCR in twenty GBS isolates, cfb gene (100%), followed by, cylE(90.47%), lmp(85.7%), bca(71.42%), rib (38%) and low frequency in bac gene (4.76%) were determined. Most of the S. agalactiae isolates produced strong biofilm in the polystyrene surface (hydrophobic), and low-level biofilm formation was found in glass tube (hydrophilic) surface. lytR is secreted protein and localized in bacterial cell wall, extra cellular membrane, and cytoplasm. In silico docking studies were performed for lytR protein with four antibiofilm compounds, including a peptide (PR39) with the docking study showed peptide has strong interaction followed by ellagic acid and interaction length is 2.95, 2.97 and 2.95 A°. In ligand EGCGO10 and O11 two atoms intract with lytR (Leu271), with binding bond affinity length is 3.24 and 3.14. The aminoacid Leu 271 is act as an impartant aminoacid, since ellagic acid and EGCG interact with same aminoacid.

Keywords: antibiotics, biofilms, clinical isolates, S. agalactiae, virulence

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Authors: Recep Keşli, Gülşah Aşık, Cengiz Demir, Onur Türkyılmaz

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Objective: Serratia species are identified as aerobic, motile Gram negative rods. The species Serratia marcescens (S. marcescens) causes both opportunistic and nosocomial infections. The genus Providencia is Gram-negative bacilli and includes urease-producing that is responsible for a wide range of human infections. Although most Providencia infections involve the urinary tract, they are also associated with gastroenteritis, wound infections, and bacteremia. The aim of this study was evaluate the antimicrobial resistance rates of S. marcescens and Providencia spp. strains which had been isolated from various clinical materials obtained from different patients who belongs to intensive care units (ICU) and inpatient clinics. Methods: A total of 35 S. marcescens and Providencia spp. strains isolated from various clinical samples admitted to Medical Microbiology Laboratory, ANS Research and Practice Hospital, Afyon Kocatepe University between October 2013 and September 2015 were included in the study. Identification of the bacteria was determined by conventional methods and VITEK 2 system (bio-Merieux, Marcy l’etoile, France) was used additionally. Antibacterial resistance tests were performed by using Kirby Bauer disc (Oxoid, Hampshire, England) diffusion method following the recommendations of CLSI. Results: The distribution of clinical samples were as follows: upper and lower respiratory tract samples 26, 74.2 % wound specimen 6, 17.1 % blood cultures 3, 8.5%. Of the 35 S. marcescens and Providencia spp. strains; 28, 80% were isolated from clinical samples sent from ICU. The resistance rates of S. marcescens strains against trimethoprim-sulfamethoxazole, piperacillin-tazobactam, imipenem, gentamicin, ciprofloxacin, ceftazidime, cefepime and amikacin were found to be 8.5 %, 22.8 %, 11.4 %, 2.8 %, 17.1 %, 40 %, 28.5 % and 5.7 % respectively. Resistance rates of Providencia spp. strains against trimethoprim-sulfamethoxazole, piperacillin-tazobactam, imipenem, gentamicin, ciprofloxacin, ceftazidime, cefepime and amikacin were found to be 10.2 %, 33,3 %, 18.7 %, 8.7 %, 13.2 %, 38.6 %, 26.7%, and 11.8 % respectively. Conclusion: S. marcescens is usually resistant to ampicillin, amoxicillin, amoxicillin/clavulanate, ampicillin/sulbactam, cefuroxime, cephamycins, nitrofurantoin, and colistin. The most effective antibiotic on the total of S. marcescens strains was found to be gentamicin 2.8 %, of the totally tested strains the highest resistance rate found against to ceftazidime 40 %. The lowest and highest resistance rates were found against gentamiycin and ceftazidime with the rates of 8.7 % and 38.6 % for Providencia spp.

Keywords: Serratia marcescens, Providencia spp., antibiotic resistance, intensive care unit

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Authors: Lucian Mocan, Cristian Matea, Flaviu A. Tabaran, Teodora Mocan, Cornel Iancu

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Introduction: Due to antibiotic resistance, systemic infections caused by Meticilin resistant Staphyloccocous Aureus (MRSA) are the main cause of millions of deaths each year. Development of new active biomolecules that are highly effective and refractory to antibiotic resistance may open new avenues in the field of antimicrobial therapy. In this research, we have focused on the development of a novel nanobiosystem with high affinity for MRSA microorganism to mediate its selective laser thermal ablation. Materials and Methods: Gold nanoparticles (15nm in diameter) linked to a specific antibody against MRSA surface were selectively delivered (at various concentrations and incubation times) and internalized into MRSA microorganism following the treatment these multidrug-resistant bacteria were irradiated using a 2w, 808 nm LASER. Results and Discussions: The post-irradiation necrotic rate ranged from 51.2% (for 1 mg/L) to 87.3% (for 50 mg/L) at 60 seconds (p<0.001), while at 30 minute the necrotic rate increased from 64.3% (1 mg/L) to 92.1% (50 mg/L), p value<0.001. Significantly lower apoptotic rates were obtained in irradiated MRSA treated with GNPs only (control) treated for 60 seconds and 30 minutes at concentrations ranging from 1 mg/L to 50 mg/L. We show here that the optimal LASER mediated the necrotic effect of MRSA after incubation with anti-MRSA-Ab was obtained at a concentration of 50 mg/L. Conclusion: In the presented research, we obtained a very efficacious pulse laser mode treatment of individual MRSA agents with minimal effects on the surrounding medium, providing highly localized destruction only for MRSA microorganism.

Keywords: MRSA, photothermolysis, antibiotic resistance, gold nanoparticles

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Authors: Jin-Hyeong Park, Hong-Seok Kim, Jin-Hyeok Yim, Young-Ji Kim, Dong-Hyeon Kim, Jung-Whan Chon, Kun-Ho Seo

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Salmonella contamination in chicken samples can cause major health problems in humans. However, not only the effects of antibiotic treatment during growth but also the impacts of poultry slaughter line on the prevalence of Salmonella in final chicken meat sold to consumers are unknown. In this study, we compared the isolation rates and antimicrobial resistance of Salmonella between antibiotic-free, conventional, conventional Korean native retail chicken meat samples and clonal divergence of Salmonella isolates by multilocus sequence typing. In addition, the distribution of extended-spectrum β-lactamase (ESBL) genes in ESBL-producing Salmonella isolates was analyzed. A total of 72 retail chicken meat samples (n = 24 antibiotic-free broiler [AFB] chickens, n = 24 conventional broiler [CB] chickens, and n = 24 conventional Korean native [CK] chickens) were collected from local retail markets in Seoul, South Korea. The isolation rates of Salmonella were 66.6% in AFB chickens, 45.8% in CB chickens, and 25% in CK chickens. By analyzing the minimum inhibitory concentrations of β -lactam antibiotics with the disc-diffusion test, we found that 81.2% of Salmonella isolates from AFB chickens, 63.6% of isolates from CB chickens, and 50% of isolates from CK chickens were ESBL producers; all ESBL-positive isolates had the CTX-M-15 genotype. Interestingly, all ESBL-producing Salmonella were revealed as ST16 by multilocus sequence typing. In addition, all CTX-M-15-positive isolates had the genetic platform of blaCTX-M gene (IS26-ISEcp1-blaCTX-M-15-IS903), to the best of our knowledge, this is the first report in Salmonella around the world. The Salmonella ST33 strain (S. Hadar) isolated in this study has never been reported in South Korea. In conclusion, our findings showed that antibiotic-free retail chicken meat products were also largely contaminated with ESBL-producing Salmonella and that their ESBL genes and genetic platforms were the same as those isolated from conventional retail chicken meat products.

Keywords: antibiotic-free poultry, conventional poultry, multilocus sequence typing, extended-spectrum β-lactamase, antimicrobial resistance

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Authors: Abdurrahim A. Elouzi, Abdurrauf M. Gusbi, Ali M. Elgerbi

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Antibiotic resistance has become a global public-health problem, thus it is imperative that new antibiotics continue to be developed. Major problems are being experienced in human medicine from antibiotic resistant bacteria. Moreover, no new chemical class of antibiotics has been introduced into medicine in the past two decades. The aim of the current study presents experimental results that evaluate the capability of bio surfactant rhamnolipid on enhancing the efficacy of hydrophobic antibiotics. Serial dilutions of azithromycin and clarithromycin were prepared. A bacterial suspension (approximately 5 X 105 CFU) from an overnight culture in MSM was inoculated into 20 ml sterile test tube each containing a serial 10-fold dilution of the test antibiotic(s) in broth with or without 200 mgL-1 rhamnolipid. The tubes were incubated for 24 h with vigorous shaking at 37°C. Antimicrobial activity in multiple antibiotic-resistant gram-negative bacteria pathogens and gram-positive bacteria were assessed using optical density technique. The results clearly demonstrated that the presence of rhamnolipid significantly improved the efficiency of both antibiotics. We hypothesized that the addition of rhamnolipid at low concentration, causes release of LPS which results in an increase in cell surface hydrophobicity. This allows increased association of cells with hydrophobic antibiotics resulting in increased cytotoxicity rates.

Keywords: hydrophobic antibiotics, biosurfactant, rhamnolipid, azithromycin, clarithromycin

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Authors: H. Akhloufi, M. Hulscher, J. M. Prins, I. H. Van Der Sijs, D. Melles, A. Verbon

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Background: A timely switch from intravenous to oral antibiotic therapy has many advantages, such as reduced incidence of IV-line related infections, a decreased hospital length of stay and less workload for healthcare professionals with equivalent patient safety. Additionally, numerous studies have demonstrated significant decreases in costs of a timely intravenous to oral antibiotic therapy switch, while maintaining efficacy and safety. However, a considerable variation in iv to oral antibiotic switch therapy criteria has been described in literature. Here, we report the development of a set of iv to oral switch criteria that are generally applicable in all hospitals. Material/methods: A RAND-modified Delphi procedure, which was composed of 3 rounds, was used. This Delphi procedure is a widely used structured process to develop consensus using multiple rounds of questionnaires within a qualified panel of selected experts. The international expert panel was multidisciplinary and composed out of clinical microbiologists, infectious disease consultants and clinical pharmacists. This panel of 19 experts appraised 6 major intravenous to oral antibiotic switch therapy criteria and operationalized these criteria using 41 measurable conditions extracted from the literature. The procedure to select a concise set of iv to oral switch criteria included 2 questionnaire rounds and a face-to-face meeting. Results: The procedure resulted in the selection of 16 measurable conditions, which operationalize 6 major intravenous to oral antibiotic switch therapy criteria. The following 6 major switch therapy criteria were selected: (1) Vital signs should be good or improving when bad. (2) Signs and symptoms related to the infection have to be resolved or improved. (3) The gastrointestinal tract has to be intact and functioning. (4) The oral route should not be compromised. (5) Absence of contra-indicated infections. (6) An oral variant of the antibiotic with good bioavailability has to exist. Conclusions: This systematic stepwise method which combined evidence and expert opinion resulted in a feasible set of 6 major intravenous to oral antibiotic switch therapy criteria operationalized by 16 measurable conditions. This set of early antibiotic iv to oral switch criteria can be used in daily practice in all adult hospital patients. Future use in audits and as rules in computer assisted decision support systems will lead to improvement of antimicrobial steward ship programs.

Keywords: antibiotic resistance, antibiotic stewardship, intravenous to oral, switch therapy

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Authors: Abdollah Jamshidi, Tayebeh Zeinali, Mehrnaz Rad, Jamshid Razmyar

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Listeria infections occur worldwide in variety of animals and man. Listeriae are widely distributed in nature. The organism has been isolated from the feces of humans and several animals, different soils, plants, aquatic environments and food of animal and vegetable origin. Listeria monocytogenes is recognized as important food-borne pathogens due to its high mortality rate. This organism is able to growth at refrigeration temperature, and high osmotic pressure. Poultry can become contaminated environmentally or through healthy carrier birds. In recent decades, prophylactic use of antimicrobial agents may be lead to emergence of antibiotic resistant organisms, which can be transmitted to human through consumption of contaminated foods. In this study, from 200 fresh chicken carcasses samples which were collected randomly from different supermarkets and butcheries, 80 samples were detected as contaminate with Listeria spp. and 19% of the isolates identified as Listeria monocytogene using multiplex PCR assay. Conventional methods were used to differentiate other species of the listeria genus. The results showed the most prevalent isolates as L. monocytogenes (48.75%). Other isolates were detected as Listeria innocua (28.75%), Listeria murrayi (20%), Listeria grayi (3.75%) and Listeria welshimeri (2.5%).The Majority of the isolates had multidrug resistance to commonly used antibiotics. Most of them were resistant to erythromycin (50%), followed by Tetracycline (44.44%), Clindamycin (41.66%), and Trimethoprim (25%). Some of them showed resistance to chloramphenicol (17.65%). The results indicate the resistance of the isolates to antimicrobials commonly used to treat human listeriosis, which could be a potential health hazard for consumers.

Keywords: listeria species, L. monocytogenes, antibiotic resistance, chicken carcass

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Authors: Chamilani Nikapitiya, Mahanama De Zoysa, Jehee Lee

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Most of the bacterial diseases are associated with high mortalities in aquaculture species and causing huge economic losses. Different approaches have been taken to prevent or control of bacterial diseases including use of vaccines, probiotics, chemotherapy, water quality management, etc. Antibiotics are widely applying as chemotherapy to control bacterial diseases, however, it has been shown that frequent use of antibiotics is favored to develop multi-drug resistance bacteria. Therefore, phages and phage encoded lytic proteins are known to be one of the most promising alternatives for antibiotics to avoid the emergence of antibiotic-resistant bacteria. We isolated and characterized the two lytic phages, namely pAh-1 and pAs-1 against pathogenic Aeromonas hydrophila and Aeromonas salmonicida, respectively. Morphological characteristics were analyzed by Transmission electron microscopy (TEM) and host strain specificities were tested with Aeromonas and other closely related bacterial strains. TEM analysis revealed that both pAh-1 and pAsm-1 are composed of an icosahedral head and a segmented tail, and we suggest that, they are new members of Myoviridae family. Genome sizes of isolated phages were estimated by restriction enzyme digestion of genomic DNA using selected endonucleases followed by agarose gel electrophoresis. Estimated genome size of pAh-1 and pAs-1 were approximately 64 Kbp and 120 Kbp, respectively. Both pAh-1 and pAs-1 have shown narrow host specificity. Moreover, protective effects of phage therapy against fish pathogenic A. hydrophila were investigated in zebrafish model. The survival rate was 40% higher when zebrafish received intra-peritoneal injection (i.p.) of pAh-1 were simultaneously challenge A. hydrophila (2 x 106 CFU/fish) compared to that without phage treatment. Overall results suggest that both pAh-1 and pAs-1 can be used as a potential phage therapy to control Aeromonas infections in aquaculture.

Keywords: Aeromonas infections, antibiotic resistance, bacteriophage, bio-control, lytic phage

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Authors: Miguel García-Ferrús, Yolanda Domínguez, M Angeles Castillo, M Antonia Ferrús, Ana Jiménez-Belenguer

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Antibiotic resistance is a growing public health concern worldwide, and it is now regarded as a critical issue within the "One Health" approach that affects human and animal health, agriculture, and environmental waste management. This concept focuses on the interconnected nature of human, animal and environmental health, and WHO highlights zoonotic diseases, food safety, and antimicrobial resistance as three particularly relevant areas for this framework. Fresh vegetables are garnering attention in the food chain due to the presence of pathogens and because they can act as a reservoir for Antibiotic Resistance Bacteria (ARB) and Antibiotic Resistance Genes (ARG). These fresh products are frequently consumed raw, thereby contributing to the spread and transmission of antibiotic resistance. Therefore, the aim of this research was to study the microbiological quality, the prevalence of ARB, and their role in the dissemination of ARG in fresh vegetables intended for human consumption. For this purpose, 102 samples of fresh vegetables (30 lettuce, 30 cabbage, 18 strawberries and 24 spinach) from different retail establishments in Valencia (Spain) have been analyzed to determine their microbiological quality and their role in spreading ARB and ARG. The samples were collected and examined according to standardized methods for total viable bacteria, coliforms, Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes and Salmonella spp. Isolation was made in culture media supplemented with antibiotics (cefotaxime and meropenem). A total of 239 strains resistant to beta-lactam antibiotics (Third-Generation Cephalosporins and Carbapenems) were isolated. Thirty Gram-negative isolates were selected and biochemically identified or partial sequencing of 16S rDNA. Their sensitivity to 12 antibiotic discs was determined using the Kirby-Bauer disc diffusion technique to different therapeutic groups. To determine the presence of ARG, PCR assays for the direct sample and selected isolate DNA were performed for main expanded spectrum beta-lactamase (ESBL)-, carbapenemase-encoding genes and plasmid-mediated quinolone resistance genes. From the total samples, 68% (24/24 spinach, 28/30 lettuce and 17/30 cabbage) showed total viable bacteria levels over the accepted standard 10(2)-10(5) cfu/g range; and 48% (24/24 spinach, 19/30 lettuce and 6/30) showed coliforms levels over the accepted standard 10(2)-10(4) cfu/g range. In 9 samples (3/24 spinach, 3/30 lettuce, 3/30 cabbage; 9/102 (9%)) E. coli levels were higher than the standard 10(3) cfu/g limit. Listeria monocytogenes, Salmonella and STEC have not been detected. Six different bacteria species were isolated from samples. Stenotrophomonas maltophilia (64%) was the prevalent species, followed by Acinetobacter pitii (14%) and Burkholderia cepacia (7%). All the isolates were resistant to at least one tested antibiotic, including meropenem (85%) and ceftazidime (46%). Of the total isolates, 86% were multidrug-resistant and 68% were ESBL productors. Results of PCR showed the presence of resistance genes to beta-lactams blaTEM (4%) and blaCMY-2 (4%), to carbapenemes blaOXA-48 (25%), blaVIM (7%), blaIMP (21%) and blaKPC (32%), and to quinolones QnrA (7%), QnrB (11%) and QnrS (18%). Thus, fresh vegetables harboring ARB and ARG constitute a potential risk to consumers. Further studies must be done to detect ARG and how they propagate in non-medical environments.

Keywords: ESBL, β-lactams, resistances, fresh vegetables.

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Authors: Wan-Ling Jiang, Hsin Chi, Jia-Lu Cheng, Ming-Fang Cheng

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Background: Investigating the risk factors for the fecal carriage of third-generation cephalosporin-resistant E.coli could contribute to further disease prevention. Previous research on third-generation cephalosporin-resistant prevalence in children in different regions of Taiwan is limited. This project aims to explore the risk factors and regional differences in the prevalence of third-generation cephalosporin-resistant and other antibiotic-resistant E. coli in the northern, southern, and eastern regions of Taiwan. Methods: We collected data from children aged 0 to 18 from community or outpatient clinics from July 2022 to May 2023 in southern, northern, and eastern Taiwan. The questionnaire was designed to survey the characteristics of participants and possible risk factors, such as clinical information, household environment, drinking water, and food habits. After collecting fecal samples and isolating stool culture with E.coli, antibiotic sensitivity tests and MLST typing were performed. Questionnaires were used to analyze the risk factors of third-generation cephalosporin-resistant E. coli in the three different regions of Taiwan. Results: In the total 246 stool samples, third-generation cephalosporin-resistant E.coli accounted for 37.4% (97/246) of all isolates. Among the three different regions of Taiwan, the highest prevalence of fecal carriage with third-generation cephalosporin-resistant E.coli was observed in southern Taiwan (42.7%), followed by northern Taiwan (35.5%) and eastern Taiwan (28.4%). Multi-drug resistant E. coli had prevalence rates of 51.9%, 66.3%, and 37.1% in the northern, southern, and eastern regions, respectively. MLST typing revealed that ST131 was the most prevalent type (11.8%). The prevalence of ST131 in northern, southern, and eastern Taiwan was 10.1%, 12.3%, and 13.2%, respectively. Risk factors analysis identified lower paternal education, overweight status, and non-vegetarian diet as statistical significance risk factors for third-generation cephalosporin-resistant E.coli. Conclusion: The fecal carriage rates of antibiotic-resistant E. coli among Taiwanese children were on the rise. This study found regional disparities in the prevalence of third-generation cephalosporin-resistant and multi-drug-resistant E. coli, with southern Taiwan having the highest prevalence. Lower paternal education, overweight, and non-vegetarian diet were the potential risk factors of third-generation cephalosporin-resistant E. coli in this study.

Keywords: Escherichia coli, fecal carriage, antimicrobial resistance, risk factors, prevalence

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Authors: Latifat Afolake Ogunfolabo, Hakeem Babafemi Ogunfolabo

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The gastrointestinal tract is densely populated with micro-organism which closely and intensively interacts with the host and ingested feed. Food borne infections are some of the major international challenges that lead to high mortality and also, antimicrobial resistance, which has been classified as a serious threat by World Health Organization. Samples of slaughtered cattle and goats intestines were collected and standard culture methods were used for bacteria isolation and identification. Minimum inhibitory concentration of commonly used antibiotic using modification of the disk diffusion method was carried out on isolates. The samples cultured were all positive to Pseudomonas aeruginosa (95% and 90%), Escherichia coli (85%), Salmonella typhi (70% and 60%), Staphylococcus aureus (75%and 100%), Micrococcus luteus (55% and35%), Bacillus macerans (60% and 5%), Bacillus cereus (25% and 20%), Clostridium perfringens (20% and 5%), Micrococcus varians (20% and 5%), Bacillus subtilis (25% and 5%), Streptococcus faecalis (40% and 25%) and Streptococcus faecium (15% and 10%) in goat and cattle respectively. Also, Proteus mirabilis (40%), Micrococcus luteus (35%), Proteus vulgaris (30%), Klebsiella aerogenes(15%) were isolated from cattle. The total coliform (13.55 x10⁵cfu/gm ± 1.77) and (20.30 x10⁵cfu/gm ± 1.27) counts were significantly higher than the total bacteria count (8.3 x10⁵cfu/gm ± 1.41) and (16.60 x10⁵cfu/gm ±0.49) for goat and cattle respectively. Selected Bacteria count of isolates showed that Staphylococcus aureus had the highest significant value (6.9 x10⁵cfu/gm ± 0.57) and (16.80 x10⁵cfu/gm ± 0.57) Escherichia coli (4.60 x10⁵cfu/gm ± 0.42) and (7.05 x10⁵cfu/gm ± 0.64) while the lowest significant value was obtained in Salmonella/Shigella (1.7 x10⁵cfu/gm ± 0.00) and (1.5 x10⁵cfu/gm ± 0.00) for goat and cattle respectively. Susceptibility of bacteria isolated from slaughtered goat and cattle intestine to commonly used antibiotics showed that the highest statistical significant value for zone of inhibition for goat was obtained for Ciprofloxacin (30.00 ± 2.25, 23.75 ± 2.49, 17.17 ± 1.40) followed by Augmentin (28.33 ± 1.22, 21. 83 ± 2.44, 16.67 ± 1.49), Erythromycin (27.75 ±1.48, 20.25 ± 1.29, 16.67 ± 1.26) while the lowest values were obtained for Ofloxacin (27.17 ± 1.89, 21.42 ± 2.19, 16.83 ± 1.26) respectively and values obtained for cattle are Ciprofloxacin (30.64 ± 1.6, 25.79 ± 1.76, 8.07 ± 11.49) followed by Augmentin (28.29 ± 1.33, 22.64 ± 1.82, 17.43 ± 1.55) Ofloxacin (26.57 ± 2.02, 20.79 ± 2.75, 16.21 ± 1.19) while the lowest values were obtained for Erythromycin (26.64 ± 1.49, 20.29 ± 1.49, 16.29 ± 1.33) at different dilution factor (10⁻¹, 10⁻², 10⁻³) respectively. The isolates from goat and cattle were all susceptible to Augmentin at the three different dilution factors. Some goat isolates are intermediate to Ciprofloxacin and Erythromycin at 10⁻² and 10⁻³, while resistance to Ciprofloxacin at 10⁻³ dilution factor. Ciprofloxacin and Ofloxacin at the dilution factors of 10⁻³ and 10⁻¹ for some cattle isolate and resistance were observed for Ofloxacin and Erythromycin at dilution of 10⁻³. These results indicate the susceptibilities and the antimicrobial resistance to commonly used antibiotic.

Keywords: antibiotic susceptibility, bacteria, cattle, goat, identification

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Authors: M. S. Raza, A. Kapil, Sonu Tyagi, H. Gautam, S. Mohapatra, R. Chaudhry, S. Sood, V. Goyal, R. Lodha, V. Sreenivas, B. K. Das

Abstract:

Background: Acute bacterial meningitis remains the major cause of mortality and morbidity. More than half of the survivors develop the significant lifelong neurological abnormalities. Diagnosis of the hospital acquired acute bacterial meningitis (HAABM) is challenging as it appears either in the post operative patients or patients acquire the organisms from the hospital environment. In both the situations, pathogens are exposed to high dose of antibiotics. Chances of getting multidrug resistance organism are very high. We have performed this experiment to find out the etiological agents of HAABM and its antibiotics susceptibility pattern. Methodology: A perspective study was conducted at the Department of Microbiology, All India Institute of Medical Sciences, New Delhi. From March 2015 to April 2018 total 400 Cerebro spinal fluid samples were collected aseptically. Samples were processed for cell count, Gram staining, and culture. Culture plates were incubated at 37°C for 18-24 hours. Organism grown on blood and MacConkey agar were identified by MALDI-TOF Vitek MS (BioMerieux, France) and antibiotic susceptibility tests were performed by Kirby Bauer disc diffusion method as per CLSI 2015 guideline. Results: Of the 400 CSF samples processed, 43 (10.75%) were culture positive for different bacteria. Out of 43 isolates, the most prevalent Gram-positive organisms were S. aureus 4 (9.30%) followed by E. faecium 3 (6.97%) & CONS 2 (4.65%). Similarly, E. coli 13 (30.23%) was the commonest Gram-negative isolates followed by A. baumannii 12 (27.90%), K. pneumonia 5 (11.62%) and P. aeruginosa 4(9.30%). Most of the antibiotics tested against the Gram-negative isolates were resistance to them. Colistin was most effective followed by Meropenem and Imepenim for all Gram-negative HAABM isolates. Similarly, most of antibiotics tested were susceptible to S. aureus and CONS. However, E. faecium (100%) were only susceptible to vancomycin and teicoplanin. Conclusion: Hospital acquired acute bacterial meningitis (HAABM) is becoming the emerging challenge as most of isolates are showing resistance to commonly used antibiotics. Gram-negative organisms are emerging as the major player of HAABM. Great care needs to be taken especially in tertiary care hospitals. Similarly, antibiotic stewardship should be followed and antibiotic susceptibility test (AST) should be performed regularly to update the antibiotic patter and to prevent from the emergence of resistance. Updated information of the AST will be helpful for the better management of the meningitis patient.

Keywords: CSF, MALDI-TOF, hospital acquired acute bacterial meningitis, AST

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Authors: Parimalakrishnan Sundararjan, Madheswaran Murugan, Dhanya Dharman, Yatindra Kumar, Sudhir Singh Gangwar, Guru Prasad Mohanta

Abstract:

Antibiotic resistance AR is a global issue, India started to redress the issues of antibiotic resistance late and it plans to have: active surveillance of microbial resistance and promote appropriate use of antibiotics. The present study attempted to achieve appropriate use of antibiotics through pharmacists’ intervention at practice point. In a quasi-experimental prospective cohort study, the cases with bacteremia from four hospitals were identified during 2015 and 2016 for intervention. The pharmacists centered intervention: active screening of each prescription and comparing with the selection of antibiotics with susceptibility of the bacteria. Wherever irrationality noticed, it was brought to the notice of the treating physician for making changes. There were two groups: intervention group and control group without intervention. The active screening and intervention in 915 patients has reduced therapeutic regimen time in patients with bacteremia. The intervention group showed the decreased duration of hospital stay 3.4 days from 5.1 days. Further, multivariate modeling of patients who were in control group showed that patients in the intervention group had a significant decrease in both duration of hospital stay and infection-related mortality. Unlike developed countries, pharmacists are not active partners in patient care in India. This unique attempt of pharmacist’ invention was planned in consultation with hospital authorities which proved beneficial in terms of reducing the duration of treatment, hospital stay, and infection-related mortality. This establishes the need for a collaborative decision making among the health workforce in patient care at least for promoting rational use of antibiotics, an attempt to combat resistance.

Keywords: antibiotics resistance, intervention, bacteremia, multivariate modeling

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Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel A. Abugri, Robertson K. Boakai, Olufemi S. Ajayi

Abstract:

The recalcitrance of pathogenic bacteria indicates that millions of people who are at risk of infection arising from chronic diseases, surgery, organ transplant, diabetes, and several other debilitating diseases present an aura of potentially untreatable illness due to resistance development. Antimicrobial resistance has successfully become a global health menace, and resistances are often acquired by bacteria through health-care-related incidence (HRI) orchestrated by multi-drug resistant (MDR) and extended drug-resistant pathogens (EDRP). To understand the mechanisms S. Typhimurium uses to resist CDB, we study the abundance of LPS modification, Ergosterols, Mysristic palmitic resistance, Oleic acid resistance of susceptible and resistant S. Typhimurium. Using qPCR, we also analyzed the expression of selected genes known for enabling resistance in S. Typhimurium. We found high abundance of LPS, Ergosterols, Mysristic palmitic resistance, Oleic acid resistance of and high expression of resistant genes in S. Typhimurium compared to the susceptible strain. LPS modification, Ergosterols, Mysristic palmitic resistance, Oleic acid and genes such as Fims, integrons, blaTEM are important indicators of resistance development of S. typhimurium.

Keywords: antimicrobials, resistance, Cannabidiol, Salmonella, blaTEM, fimA, Lipopolysaccharide, Ergosterols

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Authors: Yasir Bashawri, Vincent N. Chigor James McDonald, Merfyn Williams, Davey Jones, A. Prysor Williams

Abstract:

Resistance to antibiotics has become a threat to public health. As a result of their misuse and overuse, bacteria have become resistant to many common antibiotics. Βeta lactam (β-lactam) antibiotics are one of the most significant classes of antimicrobials in providing therapeutic benefits for the treatment of bacterial infections in both human and veterinary medicine, for approximately 60% of all antibiotics are used. In particular, some Enterobacteriaceae produce Extend Spectrum Beta Lactamases (ESBLs) that enable them to some break down multi-groups of antibiotics. CTX-M enzymes have rapidly become the most important ESBLs, with increases in mainly CTX-M 15 in many countries during the last decade. Global travel by intercontinental medical ‘tourists’, migrant employees and overseas students could theoretically be a risk factor for spreading antibiotic resistance genes in different parts of the world. Bangor city, North Wales, is subject to sudden demographic changes due to a large proportion (>25%) of the population being students, most of which arrive over a space of days. This makes it a suitable location to study the impacts of large demographic change on the presence of ESBLs. The aim of this study is to monitor the presence of ESBLs in Escherichia coli and faecal coliform bacteria isolated from Bangor wastewater treatment plant, before, during and after the arrival week of students to Bangor University. Over a five-week period, water samples were collected twice a week, from the influent, primary sedimentation tank, aeration tank and the final effluent. Isolation and counts for Escherichia coli and other faecal coliforms were done on selective agar (primary UTI agar). ESBL presence will be confirmed by phenotypic and genotypic methods. Sampling at all points of the tertiary treatment stages will indicate the effectiveness of wastewater treatment in reducing the spread of ESBLs genes. The study will yield valuable information to help tackle a problem which many regard to be the one of the biggest threats to modern-day society.

Keywords: extended spectrum β-lactamase, enterobacteriaceae, international travel, wastewater treatment plant

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Authors: Recep Kesli, Gulsah Asik, Cengiz Demir, Onur Turkyilmaz

Abstract:

Objective: Stenotrophomonas maltophilia (S. maltophilia) has recently emerged as an important nosocomial microorganism. Treatment of invasive infections caused by this organism is problematic because this microorganism is usually resistant to a wide range of commonly used antimicrobials. We aimed to evaluate clinical isolates of S. maltophilia in respect to sampling sites and antimicrobial resistant. Method: During a two years period (October 2013 and September 2015) eighteen samples collected from the intensive care unit (ICU) patients hospitalized in Afyon Kocatepe University, ANS Practice and Research Hospital. Identification of the bacteria was determined by conventional methods and automated identification system-VITEK 2 (bio-Mérieux, Marcy l’toile, France). Antibacterial resistance tests were performed by Kirby Bauer disc (Oxoid, England) diffusion method following the recommendations of CLSI. Results: Eighteen S. maltophilia strains were identified as the causative agents of different infections. The main type of infection was lower respiratory tract infection (83,4 %); three patients (16,6 %) had bloodstream infection. While, none of the 18 S. maltophilia strains were found to be resistant against to trimethoprim sulfametaxasole (TMP-SXT) and levofloxacine, eight strains 66.6 % were found to be resistant against ceftazidim. Conclusion: The isolation of S.maltophilia starains resistant to TMP-SXT is vital. In order to prevent or minimize infections due to S. maltophilia such precuations should be utilized: Avoidance of inappropriate antibiotic use, prolonged implementation of foreign devices, reinforcement of hand hygiene practices and the application of appropriate infection control practices. Microbiology laboratories also may play important roles in controlling S. maltophilia infections by monitoring the prevalence, continuously, the provision of local antibiotic resistance paterns data and the performance of synergistic studies also may help to guide appropirate antimicrobial therapy choices.

Keywords: Stenotrophomonas maltophilia, trimethoprim-sulfamethoxazole, antimicrobial resistance, Stenotrophomonas spp.

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Authors: Ibrahim Isah Lakan, Nasiru Ibrahim

Abstract:

These Phytochemical screening, Antimicrobial activities and mineral Determination of aqueous extract of Stachtarpheta indica were assessed. The result reveals the presence of flavonoids, tannins, saponins, alkaloids, glycosides and anthraquinones. The disc diffusion of aqueous extract showed Escherichia coli, 13 and antibiotic, 19 mm; Bacillus subtilis, 10 and anti –biotic, 17 mm; Klebsiller pnemuoniae , 14 and antibiotic, 24mm and Pseudmonas aeruginosa, 24 and antibiotic, 36 mm which are all comparable with the standard antibiotic cyprotomycin. The mineral content determination by flame photometer revealed that 1.25 (Na+), 0.85 (K +), 1.75 (Ca 2+) % which is a clear indication of the safety of the extract for the hypertensive patients and could be used to lower blood pressure.

Keywords: microbials, mineral, phytochemicals, stachtarpheta indica extracts

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Authors: Nourhan G. Sahly, Ahmed Moustafa, Mohamed S. Zaghloul, Tamer Z. Salem

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The gut microbiome plays important roles in the human body that includes but not limited to digestion, immunity, homeostasis and response to some drugs such as chemotherapy and immunotherapy. Its role has also been linked to radiotherapy and associated gastrointestinal injuries, where the microbial dysbiosis could be the driving force for dose determination or the complete suspension of the treatment protocol. Linking the gut microbiota alterations to different cancer treatment protocols is not easy especially in humans. However, enormous effort was exerted to understand this complex relationship. In the current study, we described the gut microbiota dysbiosis in pediatric sarcoma patients, in the pelvic region, with regards to radiotherapy and antibiotics. Fecal samples were collected as a source of microbial DNA for which the gene encoding for V3-V5 regions of 16S rRNA was sequenced. Two of the three patients understudy had experienced an increase in alpha diversity post exposure to 50.4 Gy. Although phylum Firmicutes overall relative abundance has generally decreased, six of its taxa increased in all patients. Our results may indicate the possibility of radiosensitivity or enrichment of the antibiotic resistance of the elevated taxa. Further studies are needed to describe the extent of radiosensitivity with regards to antibiotic resistance.

Keywords: combined radiotherapy and chemotherapy, gut microbiome, pediatric cancer, radiosensitivity

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Authors: Sadie Namani, Lindita Ajazaj, Arjeta Zogaj, Vera Berisha, Bahrije Halili, Luljeta Hasani, Ajete Aliu

Abstract:

Background: The outcome of bacterial meningitis is strongly related to the resistance of bacterial pathogens to the initial antimicrobial therapy. The objective of the study was to analyze the initial antimicrobial therapy, the resistance of meningeal pathogens and the outcome of adults bacterial meningitis cases. Materials/methods: This prospective study enrolled 46 adults older than 16 years of age, treated for bacterial meningitis during the years 2009 and 2010 at the infectious diseases clinic in Prishtinë. Patients are categorized into specific age groups: > 16-26 years of age (10 patients), > 26-60 years of age (25 patients) and > 60 years of age (11 patients). All p-values < 0.05 were considered statistically significant. Data were analyzed using Stata 7.1 and SPSS 13. Results: During the two year study period 46 patients (28 males) were treated for bacterial meningitis. 33 patients (72%) had a confirmed bacterial etiology; 13 meningococci, 11 pneumococci, 7 gram-negative bacilli (Ps. aeruginosa 2, Proteus sp. 2, Acinetobacter sp. 2 and Klebsiella sp. 1 case) and 2 staphylococci isolates were found. Neurological complications developed in 17 patients (37%) and the overall mortality rate was 13% (6 deaths). Neurological complications observed were: cerebral abscess (7/46; 15.2%), cerebral edema (4/46; 8.7%); haemiparesis (3/46; 6.5%); recurrent seizures (2/46; 4.3%), and single cases of thrombosis sinus cavernosus, facial nerve palsy and decerebration (1/46; 2.1%). The most common meningeal pathogens were meningococcus in the youngest age group, gram negative-bacilli in second age group and pneumococcus in eldery age group. Initial single-agent antibiotic therapy (ceftriaxone) was used in 17 patients (37%): in 60% of patients in the youngest age group and in 44% of cases in the second age group. 29 patients (63%) were treated with initial dual-agent antibiotic therapy; ceftriaxone in combination with vancomycin or ampicillin. Ceftriaxone and ampicillin were the most commonly used antibiotics for the initial empirical therapy in adults > 50 years of age. All adults > 60 years of age were treated with the initial dual-agent antibiotic therapy as in this age group was recorded the highest mortality rate (M=27%) and adverse outcome (64%). Resistance of pathogens to antimicrobics was recorded in cases caused by gram-negative bacilli and was associated with greater risk for developing neurological complications (p=0.09). None of the gram-negative bacilli were resistant to carbapenems; all were resistant to ampicillin while 5/7 isolates were resistant to cefalosporins. Resistance of meningococci and pneumococci to beta-lactams was not recorded. There were no statistical differences in the occurrence of neurological complications (p > 0.05), resistance of meningeal pathogens to antimicrobics (p > 0.05) and the inital antimicrobial therapy (one vs. two antibiotics) concerning group-ages in adults. Conclusions: The initial antibiotic therapy with ceftriaxone alone or in combination with vancomycin or ampicillin did not cover cases caused by gram-negative bacilli.

Keywords: adults, bacterial meningitis, outcomes, therapy

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Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

Abstract:

Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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Authors: Recep Keşli, Gülşah Aşık, Cengiz Demir, Onur Türkyılmaz

Abstract:

Objective: Proteus mirabilis (P. mirabilis) is one of Gram-negative pathogens in human and it causes urinary tract and nosocomial infections. P. mirabilis is susceptible to β-lactams, aminoglycosides, fluoroquinolones, and trimethoprim/sulfamethoxazole. It was aimed to investigate the resistance status to antimicrobial agents of Proteus mirabilis strains produced from samples sent to Afyon Kocatepe University, ANS Research and Practice Hospital, Microbiology Laboratory from different clinics and polyclinics during the period of 24 months. Methods: Between October 2013 and September 2015, a total of 30 Proteus were isolated from clinical samples of patients were hospitalized in intensive care units and in various departments of Afyon Kocatepe University, ANS Research and Practice Hospital. Identification of the bacteria was determined by conventional methods and VITEK 2 system (bioMérieux, France) was used additionally. Antibacterial susceptibility tests were performed by Kirby Bauer disc (Oxoid, Hempshire, England) diffusion method following the recommendations of CLSI. Results: Of the total 30 Proteus strains isolated from clinical samples, 19 from urine, 7 from wound, 4 from tracheal aspiration materials were isolated. Antimicrobial resistant for these strains were determined to 24,3% for meropenem, 26.2% for imipenem, 20.2% for amikacin 10.5% for cefepim, 33.3% for ciprofloxacin and levofloxacine, 31.6% for ceftazidime, 20% for ceftriaxone, 15.2% for gentamicin and 26.6% for amoxicillin-clavulanate, 26.2% trimethoprim-sulfamethoxale. Conclusion: In the present study, the highest number of clinical isolates of P. mirabilis were isolated from urine (63,3%), followed by the others (36,6%). The distribution of samples P. mirabilis strains to the clinics were as fallows; 16,8% intensive care unit (ICU), 29,9% polyclinics, 53,3% hospital service units The most effective antibiotic on the total of strains were found to be cefepim, the least effective antibiotics on the total of strains were found to be trimethoprim-sulfamethoxale.

Keywords: proteus mirabilis, antibiotic resistance, intensive care unit, Proteus spp.

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Authors: Avirup Chowdhury, Angus K. McFadyen, Linsey Batchelor

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Blood cultures (BCs) are an important aspect of management of the septic patient, identifying the underlying pathogen and its antibiotic sensitivities. However, while the current literature outlines indications for initial BCs to be taken, there is little guidance for repeat sampling in the following 5-day period and little information on how antibiotic use can affect the usefulness of this investigation. A retrospective cohort study was conducted using inpatients who had undergone 2 or more BCs within 5 days between April 2016 and April 2017 at a 400-bed hospital in the west of Scotland and received antibiotic therapy between the first and second BCs. The data for BC sampling was collected from the electronic microbiology database, and cross-referenced with data from the hospital electronic prescribing system. Overall, 283 BCs were included in the study, taken from 92 patients (mean 3.08 cultures per patient, range 2-10). All 92 patients had initial BCs, of which 83 were positive (90%). 65 had a further sample within 24 hours of commencement of antibiotics, with 35 positive (54%). 23 had samples within 24-48 hours, with 4 (17%) positive; 12 patients had sampling at 48-72 hours, 12 at 72-96 hours, and 10 at 96-120 hours, with none positive. McNemar’s Exact Test was used to calculate statistical significance for patients who received blood cultures in multiple time blocks (Initial, < 24h, 24-120h, > 120h). For initial vs. < 24h-post BCs (53 patients tested), the proportion of positives fell from 46/53 to 29/53 (one-tailed P=0.002, OR 3.43, 95% CI 1.48-7.96). For initial vs 24-120h (n=42), the proportions were 38/42 and 4/42 respectively (P < 0.001, OR 35.0, 95% CI 4.79-255.48). For initial vs > 120h (n=36), these were 33/36 and 2/36 (P < 0.001,OR ∞). These were also calculated for a positive in initial or < 24h vs. 24-120h (n=42), with proportions of 41/42 and 4/42 (P < 0.001, OR 38.0, 95% CI 5.22-276.78); and for initial or < 24h vs > 120h (n=36), with proportions of 35/36 and 2/36 respectively (P < 0.001, OR ∞). This data appears to show that taking an initial BC followed by a BC within 24 hours of antibiotic commencement would maximise blood culture yield while minimising the risk of false negative results. This could potentially remove the need for as many as 46% of BC samples without adversely affecting patient care. BC yield decreases sharply after 48 hours of antibiotic use, and may not provide any clinically useful information after this time. Further multi-centre studies would validate these findings, and provide a foundation for future health policy generation.

Keywords: antibiotics, blood culture, efficacy, inpatient

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Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel Abugri, Boakai Robertson, Olufemi S. Ajayi

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The emergence of multidrug resistance poses a huge risk to public health globally. Yet these recalcitrant pathogens continue to rise in incidence rate, with resistance rates significantly outpacing the speed of antibiotic development. This, therefore, presents an aura of related health issues such as untreatable nosocomial infections arising from organ transplants and surgeries, as well as community-acquired infections that are related to people with compromised immunity, e.g., diabetic and HIV patients, etc. There is a global effort to fight multidrug-resistant pathogens spearheaded by the World Health Organization, thus calling for research into novel antimicrobial agents to fight multiple drug resistance. Previously, our laboratory demonstrated that Cannabidiol (CBD) was an effective antimicrobial against Salmonella typhimurium (S. typhimurium). However, we observed resistance development over time. To understand the mechanisms S. typhimurium uses to develop resistance to Cannabidiol (CBD), we studied the abundance of bacteria lipopolysaccharide (LPS) and membrane sterols of both susceptible and resistant S. typhimurium. Using real-time quantitative polymerase chain reaction (RT-qPCR), we also analyzed the expression of selected genes known for aiding resistance development in S. typhimurium. We discovered that there was a significantly higher expression of blaTEM, fimA, fimZ, and integrons in the CBD-resistant bacteria, and these were also accompanied by a shift in abundance in cell surface molecules such as lipopolysaccharide (LPS) and sterols.

Keywords: antimicrobials, resistance, cannabidiol, gram-negative bacteria, integrons, blaTEM, Fim, LPS, ergosterols

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Authors: Afroza Parvin, Md. Mahmudul Hasan, Md. Rokunozzaman, Papon Debnath

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The effluents of textile industries have considerable amounts of heavy metals, causing potential microbial metal loads if discharged into the environment without treatment. Aim: In this present study, both lactose and non-lactose fermenting bacterial isolates were isolated from textile industrial effluents of a specific region of Bangladesh, named Savar, to compare and understand the load of heavy metals in these microorganisms determining the effects of heavy metal resistance properties on antibiotic resistance. Methods: Five different textile industrial canals of Savar were selected, and effluent samples were collected in 2016 between June to August. Total bacterial colony (TBC) was counted for day 1 to day 5 for 10-6 dilution of samples to 10-10 dilution. All the isolates were isolated and selected using 4 differential media, and tested for the determination of minimum inhibitory concentration (MIC) of heavy metals and antibiotic susceptibility test with plate assay method and modified Kirby-Bauer disc diffusion method, respectively. To detect the combined effect of heavy metals and antibiotics, a binary exposure experiment was performed, and to understand the plasmid profiling plasmid DNA was extracted by alkaline lysis method of some selective isolates. Results: Most of the cases, the colony forming units (CFU) per plate for 50 ul diluted sample were uncountable at 10-6 dilution, however, countable for 10-10 dilution and it didn’t vary much from canal to canal. A total of 50 Shigella, 50 Salmonella, and 100 E.coli (Escherichia coli) like bacterial isolates were selected for this study where the MIC was less than or equal to 0.6 mM for 100% Shigella and Salmonella like isolates, however, only 3% E. coli like isolates had the same MIC for nickel (Ni). The MIC for chromium (Cr) was less than or equal to 2.0 mM for 16% Shigella, 20% Salmonella, and 17% E. coli like isolates. Around 60% of both Shigella and Salmonella, but only 20% of E.coli like isolates had a MIC of less than or equal to 1.2 mM for lead (Pb). The most prevalent resistant pattern for azithromycin (AZM) for Shigella and Salmonella like isolates was found 38% and 48%, respectively; however, for E.coli like isolates, the highest pattern (36%) was found for sulfamethoxazole-trimethoprim (SXT). In the binary exposure experiment, antibiotic zone of inhibition was mostly increased in the presence of heavy metals for all types of isolates. The highest sized plasmid was found 21 Kb and 14 Kb for lactose and non-lactose fermenting isolates, respectively. Conclusion: Microbial resistance to antibiotics and metal ions, has potential health hazards because these traits are generally associated with transmissible plasmids. Microorganisms resistant to antibiotics and tolerant to metals appear as a result of exposure to metal-contaminated environments.

Keywords: antibiotics, effluents, heavy metals, minimum inhibitory concentration, resistance

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Authors: Maryam Beiranvand

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Multi-drug resistance in various microorganisms has increased globally in many healthcare facilities. Less effective antimicrobial activity of drug therapies for infection control becomes trouble. Since 1980, no new type of antimicrobial drug has been identified, even though combinations of antibiotic drugs have been discovered almost every decade. Between 1981 and 2006, over 70% of novel pharmaceuticals and chemical agents came from natural sources. Microorganisms have yielded almost 22,000 natural compounds. The identification of antimicrobial components from endophytes bacteria could help overcome the threat posed by multi-drug resistant strains. The project aims to analyze and identify antimicrobial peptides isolated from entophytic bacteria and their activity against multidrug-resistant Gram-negative bacteria in South Korea. Endophytic Paenibacillus polymyxa. 4G3 isolated from the plant, Gynura procumbery exhibited considerable antimicrobial activity against Methicillin-resistant Staphylococcus aureus, and Escherichia coli. The Rapid Annotations using Subsystems Technology showed that the total size of the draft genome was 5,739,603bp, containing 5178 genes with 45.8% G+C content. Genome annotation using antiSMASH version 6.0.0 was performed, which predicted the most common types of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS). In this study, diethyl aminoethyl cellulose (DEAEC) resin was used as the first step in purifying for unknown peptides, and then the target protein was identified using hydrophilic and hydrophobic solutions, optimal pH, and step-by-step tests for antimicrobial activity. This crude was subjected to C18 chromatography and elution with 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% methanol, respectively. Only the fraction eluted with 20% -60% methanol demonstrated good antimicrobial activity against MDR E. coli. The concentration of the active fragment was measured by the Brad-ford test, and Protein A280 - Thermo Fisher Scientific at the end by examining the SDS PAGE Resolving Gel, 10% Acrylamide and purity were confirmed. Our study showed that, based on the combined results of the analysis and purification. P polymyxa. 4G3 has a high potential exists for producing novel functions of polymyxin E and bacitracin against bacterial pathogens.

Keywords: endophytic bacteria, antimicrobial activity, antimicrobial peptide, whole genome sequencing analysis, multi -drug resistance gram negative bacteria

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Authors: Ji-Chan Jang

Abstract:

Tuberculosis is still major health problem because there is an increase of multidrug-resistant and extensively drug-resistant forms of the disease. Therefore, the most urgent clinical need is to discover potent agents and develop novel drug combination capable of reducing the duration of MDR and XDR tuberculosis therapy. Three reference strains H37Rv, CDC1551, W-Beijing GC1237 and six clinical isolates of MDRTB were tested to daptomycin in the range of 0.013 to 256 mg/L. Daptomycin is resistant to all tested M. tuberculosis strains not only laboratory strains but also clinical MDR strains that were isolated at different source. Daptomycin will not be an antibiotic of choice for treating infection of Gram positive atypical slowly growing M. tuberculosis.

Keywords: tuberculosis, daptomycin, resistance, Mycobacterium tuberculosis

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