Assessing antibiotic resistance of microorganisms in sanitary sewage

2014 ◽  
Vol 71 (2) ◽  
pp. 168-173 ◽  
Author(s):  
Thomas Kaeseberg ◽  
Frank Blumensaat ◽  
Jin Zhang ◽  
Peter Krebs

The release of antimicrobial substances into surface waters is of growing concern due to direct toxic effects on all trophic levels and the promotion of antibiotic resistance through sub-inhibitory concentration levels. This study showcases (1) the variation of antibiotics in sanitary sewage depending on different timescales and (2) a method to assess the antibiotic resistance based on an inhibition test. The test is based on the measurement of the oxygen uptake rate (OUR) in wastewater samples with increasing concentrations of the selected antibiotic agents. The following antibiotics were analysed in the present study: clarithromycin (CLA) was selected due to its high toxicity to many microorganisms (low EC50), ciprofloxacin (CIP) which is used to generally fight all bacteria concerning interstitial infections and doxycyclin (DOX) having a broad spectrum efficacy. Results show that CLA inhibited the OUR by approximately 50% at a concentration of about 10 mg L−1, because Gram-negative bacteria such as Escherichia coli are resistant, whereas CIP inhibited about 90% of the OUR at a concentration equal to or greater than 10 mg L−1. In the case of DOX, a moderate inhibition of about 38% at a concentration of 10 mg L−1 was identified, indicating a significant antibiotic resistance. The results are consistent with the corresponding findings from the Clinical and Laboratory Standards Institute. Thus, the presented inhibition test provides a simple but robust alternative method to assess antibiotic resistance in biofilms instead of more complex clinical tests.

2016 ◽  
Vol 88 (1) ◽  
pp. 155-163 ◽  
Author(s):  
Tiele Carvalho ◽  
Sueli Van Der Sand

Endophytic actinomycetes are promising sources of antimicrobial substances. This study evaluates the activity of metabolites produced by the endophytic actinomycete R18(6) against Gram-negative bacteria multiresistant to antimicrobials. R18(6) isolate was grown in submerged cultures under different conditions: carbon source, temperature, pH and incubation time to optimize antimicrobials production. The actinomycete grown in base medium supplemented with 1% glucose, pH 6.5 and incubation at 30 ºC for 96 h with shaking at 100 rpm, exhibited the highest activity against the used Gram-negative bacteria. Minimum inhibitory concentration (MIC) of the crude extract produced by the microorganism varied between 1/32 and 1/256. It had bactericide or bacteriostatic activity, depending on the Gram-negative organism. The active extract was stable at high temperatures, and unstable in medium containing proteolytic enzymes. Micromorphology of R18(6) was investigated by optical and scan microscopy, revealing that it was morphologically similar to the genusStreptomyces.


Author(s):  
R. H. Liss

Piperacillip (PIP) is b-[D(-)-α-(4-ethy1-2,3-dioxo-l-piperzinylcar-bonylamino)-α-phenylacetamido]-penicillanate. The broad spectrum semisynthetic β-lactam antibiotic is believed to effect bactericidal activity through its affinity for penicillin-binding proteins (PBPs), enzymes on the bacterial cytoplasmic membrane that control elongation and septation during cell growth and division. The purpose of this study was to correlate penetration and binding of 14C-PIP in bacterial cells with drug-induced lethal changes assessed by microscopic, microbiologic and biochemical methods.The bacteria used were clinical isolates of Escherichia coli and Pseudomonas aeruginosa (Figure 1). Sensitivity to the drug was determined by serial tube dilution in Trypticase Soy Broth (BBL) at an inoculum of 104 organisms/ml; the minimum inhibitory concentration of piperacillin for both bacteria was 1 μg/ml. To assess drug binding to PBPs, the bacteria were incubated with 14C-PIP (5 μg/0.09 μCi/ml); controls, in drug-free medium.


Author(s):  
Rubal C Das ◽  
Rajib Banik ◽  
Robiul Hasan Bhuiyan ◽  
Md Golam Kabir

Macrophomina phaseolina is one of the pathogenic organisms of gummosis disease of orange tree (Citrus reticulata). The pathogen was identified from the observation of their colony size, shape, colour, mycelium, conidiophore, conidia, hyaline, spore, and appressoria in the PDA culture. The crude chloroform extracts from the organism showed antibacterial activity against a number of Gram positive and Gram-negative bacteria. The crude chloroform extract also showed promising antifungal activity against three species of the genus Aspergillus. The minimum inhibitory concentration (MIC) of the crude chloroform extract from M. phaseolina against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Shigella sonnie were 128 ?gm, 256 ?gm, 128 ?gm and 64 ?gm/ml respectively. The LD50 (lethal dose) values of the cytotoxicity assay over brine shrimp of the crude chloroform extract from M. phaseolina was found to be 51.79 ?gm/ml. DOI: http://dx.doi.org/10.3329/cujbs.v5i1.13378 The Chittagong Univ. J. B. Sci.,Vol. 5(1 &2):125-133, 2010


2020 ◽  
Author(s):  
Nusrat Abedin ◽  
Abdullah Hamed A Alshehri ◽  
Ali M A Almughrbi ◽  
Olivia Moore ◽  
Sheikh Alyza ◽  
...  

Antimicrobial resistance (AMR) has become one of the more serious threats to the global health. The emergence of bacteria resistant to antimicrobial substances decreases the potencies of current antibiotics. Consequently, there is an urgent and growing need for the developing of new classes of antibiotics. Three prepared novel iron complexes have a broad-spectrum antimicrobial activity with minimum bactericidal concentration (MBC) values ranging from 3.5 to 10 mM and 3.5 to 40 mM against Gram-positive and Gram-negative bacteria with antimicrobial resistance phenotype, respectively. Time-kill studies and quantification of the extracellular DNA confirmed the bacteriolytic mode of action of the iron-halide compounds. Additionally, the novel complexes showed significant antibiofilm activity against the tested pathogenic bacterial strains at concentrations lower than the MBC. The cytotoxic effect of the complexes on different mammalian cell lines show sub-cytotoxic values at concentrations lower than the minimum bactericidal concentrations.


Author(s):  
Ganiyat Shitta ◽  
Olufunmilola Makanjuola ◽  
Olusolabomi Adefioye ◽  
Olugbenga Adekunle Olowe

Background: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the bla TEM,bla SHVand bla CTX-Mgenes. The prevalence of ESBL producing bacteria has been on the increase globally especially its upsurge among isolates from community-acquired infections. Aim: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. Material and Methods: A cross-sectional study was carried out from August 2016 –July 2017 in Osun State, Nigeria. Three hundred and sixty Gram negative bacteria recovered from clinical samples obtained from both community and healthcare associated infections were tested. They included147 Escherichia coli(40.8%), 116 Klebsiella spp(32.2%), 44 Pseudomo-nas aeruginosa(12.2%) and23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). Results: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). : Overall prevalence of ESBL producers was 41.4% with Klebsiellaspp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the blaCTX-Mgene predominating (47.0%) followed by blaTEM(30.9%) and blaSHVgene was the least, 22.1%. The blaCTX-Mgene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. Conclusion: A high prevalence of ESBL producing gram negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control spread of these pathogens should be addressed.


2020 ◽  
Vol 41 (S1) ◽  
pp. s356-s357
Author(s):  
Tomasz Kasperski ◽  
Biophage Pharma S.A. Kraków ◽  
Agnieszka Chmielarczyk ◽  
Monika Pomorska-Wesolowska ◽  
Dorota Romaniszyn ◽  
...  

Background:Acinetobacter spp are gram-negative bacteria that have emerged as a leading cause of hospital-associated infections, most often in the intensive care unit (ICU) setting. This is particularly important in Poland, where the prevalence of A. baumannii in various types of infections, including bloodstream infection (BSI), pneumonia, skin and soft-tissue infection (SSTI), and urinary tract infection (UTI) is higher than in neighboring countries. Recently, other Acinetobacter spp, including A. lwoffii or A. ursingii, have been found to be clinically relevant. In Poland, we have also observed a very rapid increase in antimicrobial resistance, significantly faster for A. baumannii than for other nosocomial pathogens. Methods: A study was conducted in 12 southern Polish hospitals, including 3 ICUs, from January 1 to December 31, 2018. Only adult hospitalized patients were included. Strains were identified using the MALDI-TOF method. Carbapenem resistance was determined using the minimum inhibitory concentration (MIC). Results: During the study, 194 strains belonging to the Acinetobacter genus were isolated. A. baumannii was the dominant species, 88.1% (n = 171), and 23 isolates (11.9%) were other Acinetobacter spp: A. ursingii (n = 5), A. lwofii (n = 4), A. haemolyticus (n = 4), A. junii (n = 3), A. radioresistens (n = 2), A. bereziniae (n = 2), and A. johnsonii (n = 2). Moreover, 15 Acinetobacter strains were collected from ICUs. The most Acinetobacter strains were isolated from SSTIs (n = 115) from non-ICU settings. Non–A. baumannii strains were also most frequently isolated from SSTIs; they constituted 11.3% of all Acinetobacter strains from this type of infection (n = 13). The total Acinetobacter prevalence was 2.6%, whereas the prevalence in the ICU setting was 7%. Acinetobacter prevalence in SSTIs was 10.4%. In pneumonia, Acinetobacter prevalence was 18.6% for ICUs (n = 13) and 2.7% for non-ICUs (n = 46). Strains from UTIs were isolated only with the non-ICU setting, and their prevalence was 0.7% (n = 14). More than half of the tested strains (52.1%) were resistant to carbapenems, but all non–A. baumannii strains were susceptible. The highest resistance to carbapenems was among strains from pneumonia cases in ICUs (58.3%) and resistance among all strains isolated from ICU was 50%. However, even higher resistance was noted among SSTI strains from non-ICUs (61.7%). Conclusions: Increasingly, more than A. baumannii, other species among Acinetobacter strains are isolated from patients hospitalized in Polish hospitals. To assess the significance of non–A. baumannii spp in clinical settings, precise species identification is needed. Therefore, the diagnostic methods used must be improved. Carbapenem-resistant A. baumannii infections are the biggest problem in pneumonia patients in ICUs and in SSTI patients in other hospital departments. Carbapenem resistance occurs in a very high percentage of A. baumannii strains; among non–A. baumannii strains it is not yet a therapeutic problem.Funding: NoneDisclosures: None


Antibiotics ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 495
Author(s):  
Masateru Nishiyama ◽  
Susan Praise ◽  
Keiichi Tsurumaki ◽  
Hiroaki Baba ◽  
Hajime Kanamori ◽  
...  

There is increasing attention toward factors that potentially contribute to antibiotic resistance (AR), as well as an interest in exploring the emergence and occurrence of antibiotic resistance bacteria (ARB). We monitored six ARBs that cause hospital outbreaks in wastewater influent to highlight the presence of these ARBs in the general population. We analyzed wastewater samples from a municipal wastewater treatment plant (MWWTP) and hospital wastewater (HW) for six species of ARB: Carbapenem-resistant Enterobacteria (CARBA), extended-spectrum β-lactamase producing Enterobacteria (ESBL), multidrug-resistant Acinetobacter (MDRA), multidrug-resistant Pseudomonas aeruginosa (MDRP), methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant Enterococci (VRE). We registered a high percentage of ARBs in MWWTP samples (>66%) for all ARBs except for MDRP, indicating a high prevalence in the population. Percentages in HW samples were low (<78%), and no VRE was detected throughout the study. CARBA and ESBL were detected in all wastewater samples, whereas MDRA and MRSA had a high abundance. This result demonstrated the functionality of using raw wastewater at MWWTP to monitor the presence and extent of ARB in healthy populations. This kind of surveillance will contribute to strengthening the efforts toward reducing ARBs through the detection of ARBs to which the general population is exposed.


Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 321
Author(s):  
Shekh Sabir ◽  
Tsz Tin Yu ◽  
Rajesh Kuppusamy ◽  
Basmah Almohaywi ◽  
George Iskander ◽  
...  

The quorum sensing (QS) system in multi-drug-resistant bacteria such as P. aeruginosa is primarily responsible for the development of antibiotic resistance and is considered an attractive target for antimicrobial drug discovery. In this study, we synthesised a series of novel selenourea and thiourea-containing dihydropyrrol-2-one (DHP) analogues as LasR antagonists. The selenium DHP derivatives displayed significantly better quorum-sensing inhibition (QSI) activities than the corresponding sulphur analogues. The most potent analogue 3e efficiently inhibited the las QS system by 81% at 125 µM and 53% at 31 µM. Additionally, all the compounds were screened for their minimum inhibitory concentration (MIC) against the Gram-positive bacterium S. aureus, and interestingly, only the selenium analogues showed antibacterial activity, with 3c and 3e being the most potent with a MIC of 15.6 µM.


2017 ◽  
Vol 83 (15) ◽  
Author(s):  
Mohammad Aminul Islam ◽  
Moydul Islam ◽  
Rashedul Hasan ◽  
M. Iqbal Hossain ◽  
Ashikun Nabi ◽  
...  

ABSTRACT Resistance to carbapenem antibiotics through the production of New Delhi metallo-β-lactamase-1 (NDM-1) constitutes an emerging challenge in the treatment of bacterial infections. To monitor the possible source of the spread of these organisms in Dhaka, Bangladesh, we conducted a comparative analysis of wastewater samples from hospital-adjacent areas (HAR) and from community areas (COM), as well as public tap water samples, for the occurrence and characteristics of NDM-1-producing bacteria. Of 72 HAR samples tested, 51 (71%) samples were positive for NDM-1-producing bacteria, as evidenced by phenotypic tests and the presence of the bla NDM-1 gene, compared to 5 of 41 (12.1%) samples from COM samples (P < 0.001). All tap water samples were negative for NDM-1-producing bacteria. Klebsiella pneumoniae (44%) was the predominant bacterial species among bla NDM-1-positive isolates, followed by Escherichia coli (29%), Acinetobacter spp. (15%), and Enterobacter spp. (9%). These bacteria were also positive for one or more other antibiotic resistance genes, including bla CTX-M-1 (80%), bla CTX-M-15 (63%), bla TEM (76%), bla SHV (33%), bla CMY-2 (16%), bla OXA-48-like (2%), bla OXA-1 (53%), and bla OXA-47-like (60%) genes. Around 40% of the isolates contained a qnr gene, while 50% had 16S rRNA methylase genes. The majority of isolates hosted multiple plasmids, and plasmids of 30 to 50 MDa carrying bla NDM-1 were self-transmissible. Our results highlight a number of issues related to the characteristics and source of spread of multidrug-resistant bacteria as a potential public health threat. In view of the existing practice of discharging untreated liquid waste into the environment, hospitals in Dhaka city contribute to the potential dissemination of NDM-1-producing bacteria into the community. IMPORTANCE Infections caused by carbapenemase-producing Enterobacteriaceae are extremely difficult to manage due to their marked resistance to a wide range of antibiotics. NDM-1 is the most recently described carbapenemase, and the bla NDM-1 gene, which encodes NDM-1, is located on self-transmissible plasmids that also carry a considerable number of other antibiotic resistance genes. The present study shows a high prevalence of NDM-1-producing organisms in the wastewater samples from hospital-adjacent areas as a potential source for the spread of these organisms to community areas in Dhaka, Bangladesh. The study also examines the characteristics of the isolates and their potential to horizontally transmit the resistance determinants. The significance of our research is in identifying the mode of spread of multiple-antibiotic-resistant organisms, which will allow the development of containment measures, leading to broader impacts in reducing their spread to the community.


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