scholarly journals Carbapenems Resistance among Gram Negative Bacteria Isolated from Poultry Samples in Gaza - Palestine

10.3823/826 ◽  
2019 ◽  
Vol 8 (3) ◽  
Author(s):  
Abdelraouf A Elmanama ◽  
Mariam R. Al-Reefi ◽  
Madleen A. Shamali ◽  
Haya I Hemaid

Background: Food is fundamental for everyone’s life. Therefore, the safety of food we consume is a priority. Gram-negative bacteria are important and common cause of human infections and could be transmitted through food handling and consumption. Carbapenemase-producing Gram-negative (CRGN) bacteria are becoming a global threat. Infections caused by CRGN are hard to cure because the carbapenems are last resort drugs for treatment. The main objective of this research is to determine the occurrence of Carbapenem-resistance among Gram-negative bacteria from poultry samples. Results:  Two hundred twenty samples (chicken litters, water, chicken feed, and intestinal content) were collected from slaughterhouses, farms, and homes from different locations in Gaza strip. Samples were cultured onto MacConkey and Blood agar plates. Gram negative isolates were identified using conventional techniques. Disk diffusion method (based on CLSI recommendations) was used to determine the antimicrobial susceptibility against 14 antimicrobials including two carbapenems (Meropenem and imipenem). Carbapenemase production was detected by the Modified Hodge Test (MHT). The Multiple Antibiotic Resistance (MAR) index for each isolate was calculated. Escherichia species were the most frequent isolates (39.5%), followed by non-lactose fermenting Enterobacteriaceae (29.5%), other lactose fermenting Enterobacteriaceae (29%). The lowest frequency was for non-fermenting Gram-negative bacilli (4.8%). Most isolates were resistant to most antimicrobial agents tested. A prominent exception was observed with meropenem, and amikacin with only 4% resistance. 41 isolates (34.7%) were resistance to imipenem. High level of intermediate results was detected for imipenem (45.2%). Among 124 isolates, 44 carbapenem-resistant (35.5%) were detected. None of the five meropenem resistant isolates and only five out of the 43 imipenem resistance isolates tested positive for carbapenemase production. Most isolates showed resistance to three or more antibiotics and are regarded as multidrug resistant strains. MDR isolates were present in 117 isolates (94.3%) with MARI index (higher than 0.3). Conclusion: Resistance to carbapenems as well as to other antimicrobials was high among GNB isolates as indicated by the MAR index. Concerned authorities should consider these alarming finding and implement an immediate monitoring program for poultry. Cross contamination, prevention measures should also be promoted and implemented.

Chemotherapy ◽  
2017 ◽  
Vol 62 (3) ◽  
pp. 194-198 ◽  
Author(s):  
Socorro Leyva-Ramos ◽  
Denisse de Loera ◽  
Jaime Cardoso-Ortiz

Background: Fluoroquinolones are widely prescribed synthetic antimicrobial agents. Quinolones act by converting their targets, gyrase and topoisomerase IV, into toxic enzymes that fragment the bacterial chromosome; the irreversible DNA damage eventually causes the killing of bacteria. Thorough knowledge of the structure-activity relationship of quinolones is essential for the development of new drugs with improved activity against resistant strains. Methods: The compounds were screened for their antibacterial activity against 4 representing strains using the Kirby-Bauer disk diffusion method. Minimal inhibitory concentration (MIC) was determined by measuring the diameter of the inhibition zone using concentrations between 250 and 0.004 μg/mL. Results: MIC of derivatives 2, 3, and 4 showed potent antimicrobial activity against gram-positive and gram-negative bacteria. The effective concentrations were 0.860 μg/mL or lower. MIC for compounds 5-11 were between 120 and 515 μg/mL against Escherichia coli and Staphylococcus aureus, and substituted hydrazinoquinolones 7-10 showed poor antibacterial activity against gram-positive and gram-negative bacteria compared with other quinolones. Conclusion: Compounds obtained by modifications on C-7 of norfloxacin with the acetylated piperazinyl, halogen atoms, and substituted hydrazinyl showed good in vitro activity - some even better than the original compound.


Author(s):  
Jyoti Diwakar ◽  
Rajesh K. Verma ◽  
Dharmendra P. Singh ◽  
Amit Singh ◽  
Sunita Kumari

Background: Carbapenemase producing multidrug-resistant organisms (i.e., MDROs) is a critical medical and public health issue globally. These bacteria are often resistant to all beta-lactam agents and are also co-resistant to other multiple classes of antimicrobial agents, leaving very few antimicrobial options.Methods: This study was carried out at UP University of medical sciences Saifai, Etawah, Uttar Pradesh, India, from January 2015 to June 2016. 110 isolates were found resistant by the Kirby Bauer’s disc diffusion method according to the CLSI guidelines. Modified Hodge test and combined disk test were performed for resistant isolates.Results: A total of 800-gram negative isolate were included in the study. 110 isolates were found resistant to imipenem by disk diffusion method. Out of these 90 (81.81%) were positive for carbapenemase production by modified Hodge test.Conclusions: We conclude that the modified Hodge test is a useful method for detection of carbapenemase production. Combined disc method is useful to detect metallo beta lactamase production.


2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S386-S386
Author(s):  
Susan M Novak-Weekley ◽  
Aye Aye Khine ◽  
Tino Alavie ◽  
Namidha Fernandez ◽  
Laxman Pandey ◽  
...  

Abstract Background Conventional antimicrobial susceptibility testing (AST) of microorganisms from positive blood cultures (PBC) can take ≥ 2 days. In order to improve the turnaround time for AST on a PBC, CLSI and EUCAST have made efforts to standardize procedures for disk diffusion (DD) direct from a PBC. Qvella Corporation (Richmond Hill, ON, Canada) has recently developed FAST-Prep, an automated centrifugal sample preparation system that rapidly delivers a Liquid Colony consisting of a purified, concentrated, viable cell suspension directly from a PBC. This study was performed to investigate the feasibility of DD AST off of a PBC using a FAST-Prep Liquid Colony. Methods Contrived PBC samples were prepared by spiking 6 species of Gram-positive and 4 species of Gram-negative bacteria (3-5 strains per species) into FA® Plus bottles and incubating in the BACT/ALERT® VIRTUO® System (bioMerieux, Durham, NC). After positivity, 3 mL of PBC was added to the FAST-Prep cartridge. After 20 minutes of processing in the FAST-Prep instrument, the Liquid Colony was removed from the cartridge and a 0.5 McFarland sample was prepared for DD AST. In parallel, the DD AST from a PBC was performed using 4 drops of PBC (CLSI direct method). Both methods were compared to conventional colony-based DD AST. After 16-18 hours of incubation zone diameters and S/I/R interpretations were determined. Categorical agreement (CA) and errors for both DD AST methods were calculated. In addition, colony plate counting was performed on 0.5 McFarland suspensions of Liquid Colony and the plate colony to determine biomass recovery and sample purity. Results CA for a FAST-Prep DD AST for Gram-positive and Gram-negative bacteria was 95.6% and 98.6%, respectively, compared to CA for CLSI DD AST of 77.2% and 81.9%, respectively. Biomass in the Liquid Colony was 7.2x108 and 1.2x109 CFU for Gram-positive and Gram-negative bacteria, respectively. Cell concentration in the 0.5 McFarland suspension of the Liquid Colony was 3.7x107 and 5.9x107 CFU/mL for Gram-positive and Gram-negative bacteria, respectively, which was similar to the concentration for the reference colony suspension. Conclusion The results support the potential role of FAST-Prep in providing a Liquid Colony for use in rapid AST. Disclosures Susan M. Novak-Weekley, PhD, D(ABMM), Qvella (Employee, Shareholder) Aye Aye Khine, PhD, Qvella (Employee, Shareholder) Tino Alavie, PhD, Qvella (Employee) Namidha Fernandez, MS, Qvella (Employee) Laxman Pandey, MS, Qvella (Employee) Abdossamad Talebpour, PhD, Qvella (Employee, Shareholder)


2012 ◽  
Vol 7 (5) ◽  
pp. 1934578X1200700 ◽  
Author(s):  
Alexis Peña ◽  
Luis Rojas ◽  
Rosa Aparicio ◽  
Libia Alarcón ◽  
José Gregorio Baptista ◽  
...  

The essential oil of the leaves of Espeletia nana Cuatrec, obtained by hydrodistillation, was analyzed by GC-MS, which allowed the identification of 24 components, which made up 99.9% of the oil. The most abundant compounds were α-pinene (38.1%), β-pinene (17.2%), myrcene (15.0%), spathulenol (4.2%), bicyclogermacrene (4.0%), α-zingiberene (4.0%), and γhimachalene (3.7%). Antibacterial activity was tested against Gram-positive and Gram-negative bacteria using the agar disk diffusion method. Activity was observed only against Gram-positive bacteria. MIC values were determined for Staphylococcus aureus ATCC 25923(200 μg/mL) and Enterococcus faecalis ATCC 29212 (600 μg/mL).


Author(s):  
Ranjit Sah ◽  
Shusila Khadka ◽  
Gentle Sunder Shrestha ◽  
Subhash Acharya ◽  
Diptesh Aryal ◽  
...  

Abstracts Background Resistance to antimicrobial agents of pathogenic bacteria has become a major problem in routine medical practices. Carbapenem resistance has long been increasing. The production of carbapenem- hydrolysing β-lactamases (carbapenamases), which include NDM, KPC, OXA-48, IMP-1 and VIM is the most common mechanism. Case presentation A 56 years old male presented with fever and mental changes with progressively decreasing sensorium for the last 3 days. He was admitted to Intensive care unit (ICU) with a diagnosis of meningoencephalitis. On day seven, he developed ventilator associated pneumonia due Klebsiella pnemoniae and Acinetobacter baumannii. He was on meropenem, but the isolates were susceptible to colistin, tigecyclin and amikacin solely. Hence, amikacin was started with addition of intravenous and nebulized colistin. Subsequently, vital signs improved with resolution of fever. However, on day 18, he developed fever once again with a drop in blood pressure. Inotropic support was maintained, and echinocandins and tigecycline were added to the regimen. Repeat blood and urine culture grew Providencia species, which were resistant to most of the drugs on phenotypic Kirby-Bauer disk diffusion method and are intrinsically resistant to colistin and tigecycline. Phenotypic detection of ESBL (combined disk method), MBL, KPCs, AmpC and co-producer were tested according to updated CLSI guideline and all were negative. But the Modified Hodges test was found to be positive. Consequenty, OXA-48 drug resistance pattern was brought into action by blank disc method according to A Tsakris et al., which revealed indentation of growth toward both EDTA and EDTA/PBA disk indicating production of OXA-48 carbapenamase. To confirm the resistance pattern we processed the isolated colonies for Xpert Carba-R (Cepheid) assay, which detected blaOXA-48 gene and confirmed the OXA-48 drug resistance pattern. Hence, the infecting organism was not susceptible to any of the antibiotics. The patient was kept under isolation and on 31th day of admission, he died of septic shock. Conclusions Carbapenamase production along with intrinsic colistin resistance in infecting bacterial pathogens can cause fatal outcomes in the resource limited countries like Nepal where new antibiotic combinations ceftazidime+ Avibactam, or aztreonam +avibactam are not available. Drug resistance patterns including OXA 48 producer should be characterized in all cases by standard phenotypic methods or by Xpert Carba-R assay and larger studies are required to know the exact burden of OXA 48 producer in Nepal.


2013 ◽  
Vol 11 (1) ◽  
pp. 45-53 ◽  
Author(s):  
Igor Stojanovic ◽  
Niko Radulovic ◽  
Vladimir Cvetkovic ◽  
Tatjana Mitrovic ◽  
Slavisa Stamenkovic

Antimicrobial activity of methanol extracts of four Parmeliaceae lichens (Hypogymnia physodes (L.) Nyl., Evernia prunastri (L.) Ach., Flavoparmelia caperata (L.) Hale and Parmelia sulcata Taylor) against a panel of microbial strains (11 Gram-positive (Enterococus sp., Bacillus subtilllis, Sarcina lutea, Micrococus luteus, Staphylococcus aureus, Clostridium sporogenes) and Gram-negative bacteria (Escherichia coli, Proteus vulgaris, Salmonela enteritidis, Pseudomonas aeruginosa, Klebsiella pneumoniae), the filamentous fungus A. niger and the yeast C. albicans) was assayed using a disk diffusion method (1 mg of the extract per disc; extracts were dissolved in methanol, 25 mg/mL). All tested extracts showed moderate antimicrobial activity. Multivariate statistical treatment (agglomerative hierarchical clustering analysis, AHC) of the obtained results allowed grouping of the samples according to their antimicrobial potential against different strains: antimicrobial profile of H. physodes and E. prunastri extracts were comparable; the similar is true for F. caperata and P. sulcata samples. In addition, based on the similarities/ dissimilarities in their susceptibility toward the tested extracts, two groups of microorganisms could be distinguished: Group I - P. vulgaris, K. pneumoniae (Gram-negative bacteria), A. niger and C. albicans; Group II - E. coli, S. enteritidis, P. aeruginosa (Gram-negative bacteria) and all of the assayed Gram-positive strains.


2019 ◽  
Vol 13 (01) ◽  
pp. 50-55
Author(s):  
Umut Safiye Say Coskun ◽  
Emel Caliskan ◽  
Asegul Copur Cicek ◽  
Halbay Turumtay ◽  
Cemal Sandalli

Introduction: The spread of Acinetobacter baumannii, resistant to most of the available antimicrobial agents, is a serious health problem. The high rate of carbapenem resistance among Acinetobacter baumannii isolates is considered as a threat to public health. In this study, we aimed to determine the antibiotic resistance and related genes in carbapenem-resistant Acinetobacter baumannii isolates. Methodology: Ninety six isolates of A. baumannii were included. Antimicrobial susceptibility was performed by Phoenix Automated System and disk diffusion method. Carbapenem resistane was characterized by scrneeing of resistance genes such as blaTEM, blaSHV, blaCTX-M1-2, blaPER, blaVEB, blaKPC, blaGES, blaNDM, blaVIM, blaIMP and blaOXA23-24-51-58 using multiplex polymerase chain reaction. Results: Resistance for the levofloxacin, gentamicin, amikacin, and tigecycline were determined as 96.9%, 93.7%, 72.9% and 45.8% respectively. Colistin was the only susceptible antibiotic against all clinical isolates. All isolates were defined as multidrug resistance and of these, 31.2% were extensively drug-resistant (sensitive only to colistin). BlaOXA-51­  and blaOXA-23 genes were detected in 100% strains while blaTEM was found in only 2% strains. There was no amplification for the blaSHV, blaCTX-M1-2, blaPER, blaVEB, blaKPC, blaGES blaNDM, blaVIM, blaIMP and blaOXA24-58 genes. Conclusions: The high frequency of blaOXA-23 and low frequency of blaTEM gene was observed that indicate prevalence of a variety of A. baumannii strains. The rates of resistance genes vary from region to region. Studies are required for the prevention and control of A. baumannii infection and to formulate the strategies of antibiotic usage.


2012 ◽  
Vol 6 (06) ◽  
pp. 489-494 ◽  
Author(s):  
Alaa Hani Al-Charrakh

Introduction: The presence of microorganisms in pharmaceuticals is undesirable because they may cause spoilage of the product and may present an infection hazard to the consumers or patients.  Methodology: A total of 102 samples of oral and topical non-sterile pharmaceutical products were collected at random from different drug houses and pharmacies in Iraq, to investigate the microbial contamination of these products. Bacterial isolates recovered from these medicaments were subjected to susceptibility testing against various antibiotics by disk diffusion method according to Clinical and Laboratory Standards (CLSI) guidelines. Results: The results revealed that the occurrence of Gram-positive bacteria was in oral and topical medicaments while Gram-negative bacteria were only detected in topical medicaments. More than 58% of Bacillus isolates were resistant to lincomycin and Bacillus mycoides isolates were resistant to beta-lactam antibiotics and trimethoprim-sulfamethoxazole. Staphylococcus spp. showed a relatively high resistance to ampicillin, amoxicillin, penicillin, tetracycline, and trimethoprim-sulfamethoxazole. S. epidermidis had the highest number of multi-resistant isolates. Furthermore, 87.5% of isolated Gram-negative rods showed high resistance to beta-lactam antibiotics and 75% of them were highly resistant to erythromycin. One isolate of Pseudomonas aeruginosa was the most resistant among all Gram-negative rod isolates. Conclusion: The high rate of resistance to antimicrobial agents of bacterial isolates recovered from oral and topical medicaments in this study may indicate a widespread antibiotic resistance among bacteria isolated from different sources, including those of anthropological and environmental origin. 


2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S93-S93
Author(s):  
Andrew Walkty ◽  
Heather Adam ◽  
Melanie Baxter ◽  
Amina Henni ◽  
Philippe Lagace-Wiens ◽  
...  

Abstract Background Inadequate empiric antimicrobial therapy for Gram-negative bacteremia is associated with adverse clinical outcomes. The purpose of this study was to evaluate the proportion of Gram-negative bacterial isolates recovered from the bloodstream of patients attending Canadian emergency rooms (ERs) that remain susceptible to commonly prescribed antimicrobials. Methods Annually from 2007 to 2018, sentinel hospitals across Canada collected bloodstream isolates from patients attending ERs as part of the CANWARD study. Susceptibility testing was performed using broth microdilution as described by CLSI (data analysis limited to Gram-negative bacteria in the top 10 pathogens), with current CLSI breakpoints applied. Extended-spectrum β-lactamase (ESBL)-producing isolates were confirmed using the CLSI disk diffusion method. Results Gram-negative bacteria among the top 10 bloodstream pathogens for patients seen at ERs across Canada were: Escherichia coli (n = 2,414), Klebsiella pneumoniae (n = 573), Pseudomonas aeruginosa (n = 211), Proteus mirabilis (n = 119), and Enterobacter cloacae (n = 114). Aggregate susceptibility of these isolates to common antimicrobials was as follows (% susceptible [S]): meropenem 99.4% S, piperacillin–tazobactam 98.5% S, gentamicin 93.3% S, ceftriaxone 88.1% S, ciprofloxacin 81.4% S, TMP-SMX 73.5% S. The most active antimicrobials evaluated vs. E. coli were meropenem (100% S), piperacillin–tazobactam (98.8% S), and ceftriaxone (93.3% S). Ceftriaxone susceptibility among E. coli isolates declined from 95.4% in 2007 to 89.8% in 2018. The average proportion of E. coli isolates that harbored an ESBL enzyme increased from 3.4% in the first three study years to 8.4% in the last three study years. The most active antimicrobials evaluated vs. K. pneumoniae isolates were meropenem (99.7% S), piperacillin–tazobactam (98.8% S), gentamicin (97.7% S), and ceftriaxone (96.9% S). Conclusion The most consistently active antimicrobials for empiric treatment of patients at Canadian ERs with Gram-negative bacteremia are meropenem and piperacillin–tazobactam. Ceftriaxone susceptibility among E. coli has declined over the last 12 years, mostly related to an increase in ESBL-producing isolates. Disclosures All authors: No reported disclosures.


Author(s):  
Maria Chatzidimitriou ◽  
Panagiota Chatzivasileiou ◽  
Georgios Sakellariou ◽  
MariaAnna Kyriazidi ◽  
Asimoula Kavvada ◽  
...  

AbstractThe present study evaluated the carbapenem resistance mechanisms of Klebsiella pneumoniae strains isolated in two Greek tertiary teaching hospitals and their susceptibility to currently used and novel antimicrobial agents.Forty-seven carbapenem resistant K. pneumoniae strains were collected in G. Papanikolaou and Ippokrateio hospital of Thessaloniki between 2016 and 2018. Strain identification and antimicrobial susceptibility was conducted by Vitek 2 system (Biomérieux France). Susceptibility against new antimicrobial agents was examined by disk diffusion method. Polymerase chain reaction (PCR) was used to detect blaKPC, blaVIM, blaNDM and blaOXA-48 genes.The meropenem–EDTA and meropenem–boronic acid synergy test performed on the 24 K. pneumoniae strains demonstrated that 8 (33.3%) yielded positive for metallo-beta-lactamases (MBL) and 16 (66.6%) for K. pneumonia carbapenemases (KPC) production. Colistin demonstrated the highest in vitro activity (87.7%) among the 47 K. pneumoniae strains followed by gentamicin (76.5%) and tigecycline (51%). Among new antibiotics ceftazidime/avibactam showed the highest sensitivity (76.6%) in all strains followed by eravacycline (66.6%). The blaKPC gene was present in 30 strains (63.8%), the blaNDM in 11 (23.4%) and the blaVIM in 6 (12.8%). The blaOXA-48 gene was not detected.Well established antimicrobial agents such as colistin, gentamicin and tigecycline and novel antibiotics like ceftazidime/avibactam and eravacycline can be reliable options for the treatment of invasive infections caused by carbapenem-resistant K. pneumoniae.


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