scholarly journals Extended-Spectrum Beta-Lactamase Gene Sequences in Gram-Negative Saprophytes on Retail Organic and Nonorganic Spinach

2011 ◽  
Vol 77 (5) ◽  
pp. 1601-1607 ◽  
Author(s):  
Eva Raphael ◽  
Lisa K. Wong ◽  
Lee W. Riley

ABSTRACTA substantial proportion of infections caused by drug-resistant Gram-negative bacteria (GNB) in community and health care settings are recognized to be caused by evolutionarily related GNB strains. Their global spread has been suggested to occur due to human activities, such as food trade and travel. These multidrug-resistant GNB pathogens often harbor mobile drug resistance genes that are highly conserved in their sequences. Because they appear across different GNB species, these genes may have origins other than human pathogens. We hypothesized that saprophytes in common human food products may serve as a reservoir for such genes. Between July 2007 and April 2008, we examined 25 batches of prepackaged retail spinach for cultivatable GNB population structure by 16S rRNA gene sequencing and for antimicrobial drug susceptibility testing and the presence of extended-spectrum beta-lactamase (ESBL) genes. We found 20 recognized GNB species among 165 (71%) of 231 randomly selected colonies cultured from spinach. Twelve strains suspected to express ESBLs based on resistance to cefotaxime and ceftazidime were further examined forblaCTX-MandblaTEMgenes. We found a 712-bp sequence inPseudomonas teessideathat was 100% identical to positions 10 to 722 of an 876-bpblaCTX-M-15gene of anE. colistrain. Additionally, we identified newly recognized ESBLblaRAHN-2sequences fromRahnella aquatilis. These observations demonstrate that saprophytes in common fresh produce can harbor drug resistance genes that are also found in internationally circulating strains of GNB pathogens; such a source may thus serve as a reservoir for drug resistance genes that ultimately enter pathogens to affect human health.

2021 ◽  
Vol 32 (1) ◽  
pp. 76-84
Author(s):  
Gideon Ikechechukwu Ogu ◽  
Faith Iguodala Akinnibosun ◽  
Odaro Stanley Imade

Abstract In Nigeria, there is still a scarcity of data on the recovery of multidrug-resistant ESBL-producing Salmonella in chicken meat. Hence this study characterized the probable multidrug-resistant extended-spectrum beta-lactamase-producing Salmonella prevalent in chilled raw chicken meat vended in Nigerian markets. Detection of Salmonella was performed by meat rinse centrifugation-plating technique. Presumptive Salmonella colonies were identified by phenotypic and 16S rRNA gene sequencing. The confirmed Salmonella isolates were tested for multidrug resistance by the Kirby Bauer disc diffusion test. Detection and confirmation of extended-spectrum beta-lactamase (ESBL) phenotypes were performed by double disc synergy and combination disc tests. PCR and DNA sequencing of the ESBL-encoding genes (bla SHV, bla TEM, and bla CTX-M) were also performed. The conserved and three-dimensional (3D) domains in ESBLs were respectively characterized by the reverse position-specific BLAST (RPS-BLAST) and Cn3D modeling tool. Of the 229 presumptive Salmonella isolates examined, 52 isolates were confirmed as Salmonella species, 46 isolates were multidrug-resistant and 41 isolates confirmed as multidrug-resistant ESBL-producing Salmonella species. The main serotypes were Salmonella enterica subsp. enterica serovar Typhimurium (35/52; 67.31%) and Salmonella enterica subsp. enterica serovar Enteritidis (17/52; 32.69%). Overall, the prevalence of chilled raw chicken meat contaminated with Salmonella was estimated at 0.17 (40/240). This value of prevalence exceeded the limits (≤ 0.1) set by the Meat Industry Guide, United Kingdom. All CTX-M, TEM, and SHV beta-lactamases produced by the Salmonella isolates were confirmed by RPS-BLAST and Cn3D modeling tool as serine-based hydrolases that consisted of two 3D domains with unique ligands such as sodium ion, formic acid, and glycerol. This study showed that multidrug-resistant ESBL-producing Salmonella was widespread in raw chicken meat vended in Nigerian markets. Thus, there is a need for relevant regulatory agencies to enforce safety.


2019 ◽  
Vol 12 ◽  
pp. 117863371988495 ◽  
Author(s):  
Adane Bitew

Background: Emergence of resistance to multiple antimicrobial agents in Non-Fermenting Gram-Negative Bacilli is a major problem to public health, as it limits drug treatment options against infections. The aim of this study was to determine the prevalence of multi-drug resistance and extended spectrum beta lactamase production in Non-Fermenting Gram-Negative Bacilli. Materials and methods: Different clinical samples were collected and processed following standard procedures. Each sample was then inoculated onto culture media. Identification, drug susceptibility testing, and extended spectrum beta lactamase production of the isolates were carried out by using the VITEK 2 compact system. Results: Among 996 clinical samples, 135 samples yielded Non-Fermenting Gram-Negative Bacilli of which Pseudomonas and Acinetobacter species were the commonest isolates. The overall drug resistance rates of Non-Fermenting Gram-Negative Bacilli were above 80% against ampicillin (89.6%), cefuroxime axetil (88.9%), nitrofurantoin (85.9%), cefalotin (84.4%), cefoxitin (83.7%), cefazolin (83.0%), and cefuroxime (83.0%). Tobramycin with a resistance rate of 19.3% was the most active antimicrobial agent. Out of 135 isolates, 81.5% were multi-drug resistant of which 13.3% were extensively drug resistant and 10.4% were pandrug resistant. Extended spectrum beta lactamase production was detected in 48.9% of the isolates. Conclusions: The spectrum of bacterial species isolated was diverse. The isolates demonstrated high level of drug resistance in different classes of antibiotics. The magnitude of multi-drug resistance and the level of extended spectrum beta lactamase production were high. Hence, further studies on multi-drug resistant and extended spectrum beta lactamase producing Non-Fermenting Gram-Negative Bacilli both in the community and in hospital setting are essential.


Author(s):  
Feleke Moges ◽  
Mucheye Gizachew ◽  
Mulat Dagnew ◽  
Azanaw Amare ◽  
Bekele Sharew ◽  
...  

Abstract Background Multidrug resistance (MDR), extended-spectrum beta-lactamase (ESBL) and carbapenemase-producing Gram-negative bacteria (GNB) has become a public health threat worldwide. This threat is worse in developing countries where there is high infectious disease burden and spread of antimicrobial resistance co-exist. The aim of the present study was, therefore, to assess MDR, ESBL and carbapenemase producing GNB from patients attending three selected referral hospitals in Amhara region. Methods A cross-sectional study was conducted from December 2017- April 2018 at the University of Gondar Comprehensive Specialized Hospital, Dessie Referral Hospital and Debre Markos Referral Hospital of Amhara national regional state. A total of 833 study subjects were recruited using a convenient sampling technique. Clinical samples such as blood, urine, stool, wound, abscess, ear discharge, nasal discharge, cervical discharge and body fluid specimens were aseptically collected. Culturing for identification of bacteria and determination of drug susceptibility testing were done following standard microbiological techniques. Selected MDR isolates were phenotypically assessed for ESBL and carbapenemase production. Results Of the 833 clinical samples cultured for bacterial growth, 141 (16.9%) were positive for GNB. The most common GNB identified were E. coli 46 (32.6%), Klebsiella spp. 38 (26.5%) and Proteus spp. 13 (9.2%). The overall MDR prevalence was 121 (85.8%). Among the total isolates, 137 (97.2%) were resistant to ampicillin followed by cotrimoxazole 115 (81.6%), amoxicillin-clavulanic acid 109 (77.3%), cefixime 99 (70.2%), cefepime 93 (66.0%) and tetracycline 91 (64.5%). The extended-spectrum beta-lactamase producing GNB were 69/124 (55.6%). Of which Klebsiella spp. 19 (15.3%) and E. coli 17 (13.7%) were common ESBL producers. Carbapenemase-producing isolates were 8/51(15.7%). Of which Enterobacter, Klebsiella and E. coli were common carbapenemase producers. Conclusion and recommendation Multi-drug resistance and ESBL producing isolates in the present study were high. E. coli and Klebsiella spp. were the most common ESBL producing GNB. Klebsiella spp., Enterobacter spp., E. coli and Citrobacter spp. were typical carbapenemase-producing isolates. Continuous monitoring, antibiotic stewardship and molecular detection of the gene responsible for drug resistance are important means to reduce the spread of drug-resistant pathogens.


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.


KYAMC Journal ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 171-175
Author(s):  
Tania Rahman ◽  
Momtaz Begum ◽  
Sharmeen Sultana ◽  
SM Shamsuzzaman

Background: In recent years, Extended-spectrum beta-lactamase (ESBL) producing microorganisms have complicated treatment of infections due to resistance of ESBL producing strains to a wide range of antimicrobials. Objective: Target of this study was to determine the prevalence of ESBL producing gramnegative bacteria in neonatal sepsis cases and to reveal the antimicrobial susceptibility pattern of those isolated ESBL producers. Materials and Methods: This cross sectional study was carried out in Dhaka Medical College Hospital (DMCH) over a period of 12 months from January to December in 2016. Following isolation and identification of gram-negative bacteria from blood samples of suspected septicemic neonates, antimicrobial susceptibility test was performed by Kirby Bauer disk-diffusion method and ESBL producers were detected by Double Disk Synergy (DDS) test. Results: Among 52 Gram-negative bacteria isolated from 106 blood samples, 34.61% ESBL producers were detected and Enterobacter spp. (45%) was predominant followed by Klebsiella pneumoniae (33.33%). None of the ESBL producers was resistant to colistin and tigecycline. All ESBL producing Acinetobacter baumannii, 77.78% and 66.67% of ESBL producing Enterobacter spp and Klebsiella spp. respectively showed resistance to meropenem. All ESBL producers were resistant to piperacillintazobactam. Conclusion: Appropriate measures should be taken to prevent the spread of ESBL producing strains by combining strategies for infection prevention, control and rational use of antibiotics. KYAMC Journal Vol. 11, No.-4, January 2021, Page 171-175


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