Phenotypic and Molecular Characterization of Staphylococcaceae and Enterobacteriaceae Species Isolated from Smoked, Dried, and Braised Fish Marketed in Ouagadougou

Background: To preserve its nutritional properties, fish must have good sanitary quality. The objective of this study was to investigate some pathogens contaminating smoked, dried, and braised fish marketed in Ouagadougou. Methodology: Potential pathogens of Enterobacteriaceae and Staphylococcus were screened in eight (8) species of processed fish. The investigation of the germs was carried out following the normative methods of microbiology. The identities of the strains were determined by API 20 E (BioMerieux S.A., France) and API STAPH (BioMerieux S.A., France) kits for Enterobacteriaceae and Staphylococcus species respectively. The uidA gene profile in Escherichia coli isolates was determined by simplex PCR. The identity of Staphylococcus aureus was confirmed by amplification of specific 23S rDNA regions and nuc gene profile with PCR. Results: A total of 235 fish samples were analyzed. A diversity of Enterobacteriaceae and Staphylococcus was detected. Twenty species of Enterobacteriaceae were identified among which, the most frequent were Escherichia coli, Salmonella sp, Raoultella ornithinolytica and Serratia odorifera, respectively in 22.6%, 4.3%, 28.9%, 17.4% of the samples analyzed. However, eleven species of Staphylococcus were identified among which, Staphylococcus xylosus, Staphylococcus aureus, Staphylococcus lugdunensis, and Staphylococcus sciuri were the most frequent with respective percentages of 47.7%, 23.4%, 12.8% and 10.6% of samples. For all the samples, the species frequently isolated were: Raoultella ornithinolytica, Escherichia coli, Serratia odorifera, Staphylococcus aureus, Staphylococcus xylosus, and Staphylococcus lugdunensis. The uidA gene specific to Escherichia coli was detected in 82.85% of strains (29/35). Amplification of the specific 23S rDNA region using staur primers was observed in 98% (49/50) of the isolated Staphylococcus aureus strains and the nuc gene was detected in 86% of Staphylococcus aureus strains. Conclusion: The isolated bacteria are potential pathogens involved in foodborne illnesses and intoxications. Effective sanitary safety systems must be implemented to guarantee the sanitary quality of fish supplied to consumers.

Phenotypic Characterization of Carbapenemase-Producing Enterobacteriaceae Strains in a Referral Teaching Hospital in Yaoundé, Cameroon

Background: Carbapenemase-producing Enterobacteriaceae (CPE) are an important and increasing threat to global health. They are nowadays more encountered routinely in hospitals and cause high morbidity and mortality due to limited therapeutic alternatives. This study sought to determine the prevalence of CPE in Yaoundé teaching hospital, Cameroon, and the associated risk factors. Materials and Method: To achieve this goal, a descriptive cross-sectional study coupled to an analytical component with consecutive collection of Enterobacteria strains was carried out during a three-month period (from 27th July to 24th October 2018) in the University Teaching Hospital of Yaoundé, Cameroon. The oxidase and biochemical identification tests using a miniaturized Api 20 E system were performed on colonies grown on Eosin Methylene Blue (EMB) medium and subcultured on nutrient agar. Drug susceptibility testing was carried out according to the Antibiogram Committee of the French Society of Microbiology (CA-SFM 2018.V.2.0). The detection of carbapenemase production was performed by the CA-SFM 2018 algorithm for the screening of carbapenemase-producing enterobacteriaceae and its classification by inhibitory synergy tests. Results: Out of the 104 isolates, Escherichia coli (50%) was the most prevalent species, followed by Klebsiella pneumoniae (37.5%) and Citrobacter frendii (12.5%). Drugs susceptibility patterns showed a high resistance to penicillins group (97.4% to amoxicillin), cephalosporins (68.4% to cefotaxim, 58.1% to cefixim, 60.7% to ceftazidim, 57.1% of cefoxitin) and aztreonam (55.7%). However, 11.9% carbapenems related resistance was noticed: 14.4% to imipenem, 13.8% to ertapenem and 7.5% to meropenem. Numerous co-resistance to quinolones (65.8%), fluoroquinolones (49.6%), aminoglycosides (49.6%) and cotrimoxazole (71.8%) were also observed. From 104 isolates, AmpC production represented 23.08% (25/104) and 36.54% (38/104) were ESBL-isolates. The overall prevalence of CPE was 25% (26/104) with K.pneumoniae predominant (61.53%). Besides, Class A and class B carbapenemase were mainly produced with respectively 20% (21/104) and 5% (5/104). Univariate analysis revealed a significant association of carbapenemase production to Klebsiella pneumoniae (p = 0.01), ESBL and AmpC production ((P = 0.01 and P = 0.001 respectively) while that association was only significant to Klebsiella spp (p = 0.04) and AmpC production (p = 0.02) in multivariate analysis. Conclusion: The multi-resistance of Enterobacteriaceae to antibiotics in Cameroon has considerably increased. More attention should be paid to those bacteria to stall antimicrobial resistance spread.

Risk of transmission of carbapenem-resistant Enterobacteriaceae and related “superbugs” during gastrointestinal endoscopy

To evaluate the risk of transmission of carbapenem-resistant Enterobacteriaceae(CRE) and their related superbugs during gastrointestinal(GI) endoscopy. Reports of outbreaks linked to GI endoscopes contami-nated with different types of infectious agents, includ-ing CRE and their related superbugs, were reviewed. Published during the past 30 years, both prior to and since CRE's emergence, these reports were obtained by searching the peer-reviewed medical literature(via the United States National Library of Medicine's "MEDLINE" database); the Food and Drug Administration's Manu-facturer and User Facility Device Experience database, or "MAUDE"; and the Internet(via Google's search engine). This review focused on an outbreak of CRE in 2013 following the GI endoscopic procedure known as endoscopic retrograde cholangiopancreatography, or ERCP, performed at "Hospital X" located in the sub-urbs of Chicago(IL; United States). Part of the largest outbreak of CRE in United States history, the infection and colonization of 10 and 28 of this hospital's patients, respectively, received considerable media attention and was also investigated by the Centers for Disease Con-trol and Prevention(CDC), which published a report about this outbreak in Morbidity and Mortality WeeklyReport(MMWR), in 2014. This report, along with the results of an independent inspection of Hospital X's in-fection control practices following this CRE outbreak, were also reviewed. While this article focuses primar-ily on the prevention of transmissions of CRE and their related superbugs in the GI endoscopic setting, some of its discussion and recommendations may also apply to other healthcare settings, to other types of flexible endoscopes, and to other types of transmissible infec-tious agents. This review found that GI endoscopy is an important risk factor for the transmission of CRE and their related superbugs, having been recently as-sociated with patient morbidity and mortality following ERCP. The CDC reported in MMWR that the type of GI endoscope, known as an ERCP endoscope, that Hospi-tal X used to perform ERCP in 2013 on the 38 patients who became infected or colonized with CRE might be particularly challenging to clean and disinfect, because of the complexity of its physical design. If performed in strict accordance with the endoscope manufacturer's labeling, supplemented as needed with professional organizations' published guidelines, however, current practices for reprocessing GI endoscopes, which include high-level disinfection, are reportedly adequate for the prevention of transmission of CRE and their related superbugs. Several recommendations are provided to prevent CRE transmissions in the healthcare setting. CRE transmissions are not limited to contaminated GI endoscopes and also have been linked to other reusable flexible endoscopic instrumentation, including broncho-scopes and cystoscopes. In conclusion, contaminated GI endoscopes, particularly those used during ERCP, have been causally linked to outbreaks of CRE and their related superbugs, with associated patient morbidity and mortality. Thorough reprocessing of these complex reusable instruments is necessary to prevent disease transmission and ensure patient safety during GI endos-copy. Enhanced training and monitoring of reprocessing staffers to verify the proper cleaning and brushing of GI endoscopes, especially the area around, behind andnear the forceps elevator located at the distal end othe ERCP endoscope, are recommended. If the ERCPendoscope features a narrow and exposed channel thathouses a wire connecting the GI endoscope's controhead to this forceps elevator, then this channel's com-plete reprocessing, including its flushing with a deter-gent using a procedure validated for effectiveness, is also emphasized.

Bacteriophage therapy against Enterobacteriaceae

The Enterobacteriaceae are a class of gram-negative facultative anaerobic rods, which can cause a variety of diseases, such as bacteremia, septic arthritis, endocarditis, osteomyelitis, lower respiratory tract infections, skin and soft-tissue infections, urinary tract infections, intra-abdominal infections and ophthalmic infections, in humans, poultry, animals and fish. Disease caused by Enterobacteriaceae cause the deaths of millions of people every year, resulting in enormous economic loss. Drug treatment is a useful and efficient way to control Enterobacteriaceae infections. However, with the abuse of antibiotics, drug resistance has been found in growing number of Enterobacteriaceae infections and, as such, there is an urgent need to find new methods of control. Bacteriophage therapy is an efficient alternative to antibiotics as it employs a different antibacterial mechanism. This paper summarizes the history of bacteriophage therapy, its bacteriallytic mechanisms, and the studies that have focused on Enterobacteriaceae and bacteriophage therapy.

Treatment of sepsis: What is the antibiotic choice in bacteremia due to carbapenem resistant Enterobacteriaceae?

Sepsis is one of the major challenges of today. Although gram-positive bacteria related infections are more prevalent in hospital setting, the highest mortality rate is associated with gram-negative microorganisms especially Enterobacteriaceae. Enterobacteriaceae, including Escherichia coli, Klebsiella spp., Proteus spp., Enterobacter spp. and Serratia spp. Resistance to β-lactams in Enterobacteriaceae is primarily attributed to the production of B-lactamase enzymes with subsequent antibiotic hydrolysis and to a lesser extent by alteration of efflux pump or porins expression. Carbapenem resistant Enterobacteriaceae(CRE) and Acinetobacter baumannii are the most notorious pathogens due to the high incidence of morbidity and mortality especially in the immunocompromised patients in the intensive care unit. The most appropriate antimicrobial therapy to treat CRE is still controversial. Combination therapy is preferred over monotherapy due to its broad-spectrum coverage of micro-organisms, due to its synergetic effect and to prevent development of further resistance. Current suggested therapies for CRE resistance as well as promising antibiotics that are currently under investigation for winning the war against the emerging CRE resistance are reviewed and discussed.

Characterization of NDM-5-producing Enterobacteriaceae isolates from retail grass carp(Ctenopharyngodon idella) and evidence of blaNDM-5-bearing IncHI2 plasmid transfer between ducks and fish

We aimed to characterize NDM-5-producing Enterobacteriaceae from aquatic products in Guangzhou,China.A total of 196 intestinal samples of grass carp collected in 2019 were screened for carbapenemase genes.Characterization of bla_(NDM-5) positive isolates and plasmids was determined by antimicrobial susceptibility testing,conjugation experiments,Illumina HiSeq,and Nanopore sequencing.One Citrobacter freundii and six Escherichia coli strains recovered from seven intestinal samples were verified as bla_(NDM-5) carriers(3.57%,7/196).The bla_(NDM-5) genes were located on the lncX3(n=5),lncHI2(n=1),or lncHI2-lncF(n=1)plasmids.All bla_(NDM-5)-bearing plasmids were transferred by conjugation at frequencies of~10^(-4)-10^(-6).Based on sequence analysis,the lncHI2 plasmid pHNBYF33-1 was similar to other bla_(NDM-5)-carrying lncHI2 plasmids deposited in GenBank from Guangdong ducks.In all lncHI2 plasmids,bla_(NDM-5)was embedded in a novel transposon,Tn7057(IS3000-△ISAba125-IS5-△ISAba125-bla_(NDM-5)-bleMBL-trpF-tat-△dct-IS26-△umuD-△ISKox3-IS3000),which was identical to the genetic structure surrounding bla_(NDM-5)found in some IncX3 plasmids.The lncHI2-lncF hybrid plasmid pHNTH9F11-1 was formed by homologous recombination of the bla_(NDM-5)-carrying lncHI2 plasmid and a heavy-metal-resistant IncF plasmid through△Tn1721 To the best of our knowledge,this is the first report on the characterization of bla_(NDM-5)-bearing plasmids in fish in China.The lncHI2 plasmid pHNBYF33-1 may be transmitted from ducks,considering the common duck-fish freshwater aquaculture system in Guangdong.Tn7051 is likely responsible for the transfer of bla_(NDM-5) from lncX3 to lncHI2 plasmids in Enterobacteriaceae,resulting in the expansion of transmission vectors of bla_(NDM-5).

Genomic and Phenotypic Diversity of Carbapenemase-Producing Enterobacteriaceae Isolates from Bacteremia in China: A Multicenter Epidemiological, Microbiological, and Genetic Study

Carbapenemase-producing Enterobacteriaceae(CPE) isolates are recognized as one of the most severe threats to public health. However, the population structure and genetic characteristics of CPE isolates among bloodstream infections(BSIs) are largely unknown. To address this knowledge gap, in this study,we included patients with clinically significant BSIs due to Enterobacterales isolates, recruited from 26 sentinel hospitals in China(2014–2015). CPE isolates were microbiologically and genomically characterized,including their susceptibility profiles, molecular typing, phylogenetic features, and genetic context analysis of carbapenemase-encoding genes. Of the 2569 BSI Enterobacterales isolates enrolled, 42(1.6%) were carbapenemase-positive. Moreover, among the 2242 investigated isolates, 1111(49.6%) extendedspectrum β-lactamase(ESBL)-producing isolates were identified in Escherichia coli(E. coli), Klebsiella pneumoniae(K. pneumoniae), Proteus mirabilis(P. mirabilis), and Klebsiella oxytoca. Whole genome sequencing analysis showed the clonal spread of K. pneumoniae carbapenemase(KPC)-2-producing K. pneumoniae sequence type(ST) 11 and New Delhi metallo-β-lactamase(NDM)-5-producing E. coli ST167 in our collection. Plasmid analysis revealed that carbapenemase-encoding genes were located on multiple plasmids. A high prevalence of biofilm-encoding type 3 fimbriae clusters and yesiniabactin-associated genes was observed in K. pneumoniae isolates. This work demonstrates the high prevalence of ESBLs and the wide dissemination of CPE among BSI isolates in China, which represent real clinical threats. Moreover, our findings first illustrate a more comprehensive genome scenario of CPE isolates among BSIs. The clonal spread of KPC-2-producing K. pneumoniae ST11 and NDM-5-producing E. coli ST167 needs to be closely monitored.

bla_(CTX-M),bla_(TEM),and bla_(SHV) in Enterobacteriaceae from North-Indian tertiary hospital:high occurrence of combination genes

Objective:To delineate the frequency of occurrence of bla_(CTX-M),bla_(TEM),and bla_(SHV) in Enterobacteriaceae from North-Indian tertiary hospital.Methods:A random collection of a subset of 45 Escherichia coli(E.coli) and 28 Klebsiella pneumoniae(K.pneumoniae) that was resistant to a third generation cephalosporin and obtained during 2007-2008 was selected for detailed screening for bla_(CTX-M),bla_(TEM),and bla_(SHV) by monoplex PCRs.The isolates demonstrating the presence of bla_(CTX-M) alleles were characterized for the specific CTX-M-genogroup by using a multiplex PCR.Results:Resistance to cefoperazone,ceftazidime,ceftriaxone,cefotaxime, cefoxitin and piperacillin was 100%each in K.pneumoniae isolates,whereas these resistancerates for E.coli isolates were 93.1%,83.8%,91.9%,93.6%,97.3%and 97.1%,respectively. Concomitant resistance to aminoglycosides,quinolones and aztreonam was also noticed.Presence of any of the bla genes(bla_(CTX-M),bla_(TEM),and bla_(SHV)) was noticed in a total of 28(38.4%) isolates of the 73 isolates studied.Many isolates demonstrated occurrence of these genes in various combinations.bla_(CTX-M),bla_(TEM),and bla_(SHV) were noticed in 28.8%,10.9%and 13.7%isolates, respectively.Multiplex PCR in bla_(CTX-M) harboring isolates demonstrated the presence of CTX-MGenogroup -1 and sequencing for the specific CTX-M-type revealed presence of CTX-M-15 type. RAPD typing showed wide diversity in isolates.Conclusions:This is amongst the premier report describing the simultaneous occurrence of blo_(TEM),bla_(SHV),and bla_(ampC) in Indian Enterobacteriaceae and that wider dissemination of these genes,as demonstrated by diversity of isolates,raises concern and emphasizes a need for extensive search for the presence of these gene pools in Indian subcontinent.

Increasing secondary bacterial infections with Enterobacteriaceae harboring bla_(CTX-M-15) and bla_(CMY-6) in patients with bronchogenic carcinoma:an emerging point of concern

Objective:To look for secondary bacterial infections in bronchogenic carcinoma（BCA） with resistant organisms harboring bla genes considering the paucity of relevant studies.Methods: A total of 137 confirmed cases of BCA and 34 healthy volunteers were studied for the occurrence and prevalence of blaCTX-M and and blaAmpC harboring-enterobacteriaceae.A subset of these patients（n=69） was previously reported for the secondary infection with the Aspergillus species. Bronchoalveolar lavages（BAL） were subjected for bacterial and fungal cultures and the bacterial isolates were screened by multiplex PCRs for the presence of blaCTX-M and blaAmpC.The isolates were also screened for the association of insertion sequence（IS26） by PCR and characterized by RAPD for any clonal relatedness.Results:A total of 143 bacterial isolates were obtained from 137 BAL specimens of BC,patients.The Enterobacteriaceae-isolates were multidrug-resistant showing concomitant resistance to fluoroquinolones and aminoglycosides.Both blaCTX-M and blaAmpC of CIT family were detected in 77.4%and 27.4%isolates,respectively.Sequencing revealed the presence of blaCTX-M-15 and blaCMY-6.Twenty one percent of the isolates were simultaneously harboring blaampC and blaCTX-M-15.IS26 PCR and RAPD typing revealed the presence of diverse bacterial population but no predominant clone was identified.The present study also suggests strong association of aspergillosis with lung cancer and further strengthens the potential use of non-validated serological tests suggested earlier.Conclusions:We emphasize that all patients of bronchogenic carcinoma should also be screened for secondary bacterial infections,along with secondary fungal infections,so as to introduce early and specific antimicrobial therapy and to prevent unwanted deaths.

Spontaneous bacterial peritonitis due to carbapenemase-producing Enterobacteriaceae:Etiology and antibiotic treatment

Carbapenem antibiotics were first introduced in the 1980s and have long been considered the most active agents for the treatment of multidrug-resistant gramnegative bacteria.Over the last decade,carbapenem-resistant Enterobacteriaceae(CRE)have emerged as organisms causing spontaneous bacterial peritonitis.Infections caused by CRE have shown a higher mortality rate than those caused by bacteria sensitive to carbapenem antibiotics.Current antibiotic guidelines for the treatment of spontaneous bacterial peritonitis are insufficient,and rapid deescalation of empiric antibiotic treatment is not widely recognized.This review summarizes the molecular characteristics,epidemiology and possible treatment of spontaneous bacterial peritonitis caused by CRE.

Abundance of Enterobacteriaceae in the colon mucosa in diverticular disease

AIM To compare gut bacterial diversity and amount of Enterobacteriaceae in colonic mucosa between patients with and without diverticular disease(DD).METHODS Patients in a stable clinical condition with planned elective colonoscopy were included. Blood samples and colon mucosa biopsies were collected at the colonoscopy. Study questionnaires including questions about gastrointestinal symptoms were completed by the patients and physicians. DNA from mucosa samples was isolated and the amount of Enterobacteriaceae was estimated using PCR assay. Terminal restriction frag mentlength polymorphism was applied to assess microbial diversity. Diversity was estimated by calculations of richness(number of terminal restriction fragments) and Shannon-Wiener and Simpson's indices. RESULTS A total of 51 patients were included, 16 patients with DD [68(62-76) years] and 35 controls [62(40-74) years] without any diverticula. Patients with DD had significantly higher levels of Enterobacteriaceae than those without DD(P = 0.043), and there was an inverse relationship between the amount of Enterobacteriaceae and the Simpson's index(rs =-0.361, P = 0.033) and the Shannon-Wiener index(rs =-0.299, P = 0.081). The Simpson's index(P = 0.383), Shannon-Wiener index(P = 0.401) or number of restrictions fragments(P = 0.776) did not differ between DD and controls. The majority of patients experienced gastrointestinal symptoms, and 22 patients(43.1%) fulfilled the criteria for irritable bowel syndrome, with no difference between the groups(P = 0.212). Demography, socioeconomic status, lifestyle habits, inflammatory biomarkers, or symptoms were not related to the amount of Enterobacteriaceae or bacterial diversity. CONCLUSION Patients with DD had higher amount of Enterobacteriaceae in the colon mucosa compared to patients without diverticula.

Isolation, Characterization and Antimicrobial Resistance Patterns of Lactose-Fermenter Enterobacteriaceae Isolates from Clinical and Environmental Samples

The lactose-fermenter Enterobacteriaceae are the most frequent cause of clinical infection in our country. The objective of this study was to isolate and identify the most common lactose-fermenter Enterobacteriaceae from clinical samples, including urine, blood, wounds, and sputum, obtained from the local hospital and from environmental samples from a chicken farm, agriculture soil, and water from the Tigris River in Baghdad City. The study also aimed at establishing the antibiotic resistance patterns of the isolated bacteria. A total of 155 bacterial isolates were identified from 10 genera according to the Vitek 2 system. The most common bacterial isolates from the clinical and environmental samples were Escherichia coli and Klebsiella pneumoniae, respectively. The antibiotic resistance patterns showed that all clinical and environmental isolates were multidrug resistant to β-lactam (except carbapenems) drug and aminoglycosides and more sensitive to carbapenems.

Antimicrobial Resistance among Enterobacteriaceae Found in Chicken and Cow Droppings and Their Public Health Importance

Introduction: The recent surge in the number of antimicrobial resistant cases from hospitals and communities has created a need to study the points and sources of exposure to certain bacteria and determine their susceptibility to commonly used antibiotics. This study aimed at identifying and screening for drug-resistant Enterobacteriaceae isolated from chicken droppings and cow dungs in Onitsha, Anambra state, in the South-Eastern part of Nigeria. Methods: This is a cross-sectional descriptive study which included 50 chickens and 50 cow dung samples collected from five poultry houses and cow ranches respectively using sterile swab sticks. The samples were transported to the laboratory and processed following standard microbiological protocols. Isolates in the samples were recovered using MacConkey Agar, Eosin Methylene Blue Agar and Salmonella-Shigella Agar following standard microbiological procedures and then identified/characterized biochemically using commercial API 20E identification kits following the standard manufacturer’s protocol. Isolates were subjected to antibiotic susceptibility testing on Muller Hinton Agar using Kirby Bauer double-disc diffusion technique. The multiple antibiotics resistance index was determined as well. Isolates with reduced susceptibility to Ceftazidime were screened for extended spectrum beta-lactamase, AmpC- and metallo-beta-lactamase-production using Rosco Diagnostic kit. Results: Sixty-two (100%) Gram-negative bacteria were isolated from a total of 100 samples collected from both sites, out of which 43 (69.4%) are Enterobacteriaceae. A total of 30/43 (69.8%) Enterobacteriaceae including K. pneumoniae, S. enteritica, S. odorifera, E. coli, K. intermediate, P. stuartii, E. aerogenes, P. penneri, P. mirabilis and C. braakii were recovered from chicken droppings, whereas 13/43 (30.2%) Enterobacteriaceae including K. pneumoniae, S. enteritica, S. odorifera, E. coli, K. intermediate, P. stuartii, E. aerogenes, P. penneri, P. mirabilis and C. braakii were recovered from cow dungs. Two (12.5%) different isolates demonstrated metallo-beta-lactamase and cephalosporinase (AmpC) production. The isolates were susceptible to six antibiotics tested except Augmentin and Nitrofurantoin where the resistance is 100% and 85% respectively while Ceftriaxone and Ofloxacin had the best antibacterial activity against the isolates from both sites. Conclusion: The bacteria of public health importance isolated from these sites and their antibiogram profile have shown the need for proper monitoring and management of animal wastes in order to mitigate the threat to human health in the spirit of One Health as well as contribute to the fight against antibiotic resistance.

Comparative study on occurrence of class A and class Cβ-lactamase genes and their co-occurrence in Indian Enterobacteriaceae during years 2009 and 2010

Objective:To determine the occurrence of class A and class Cβ-Iactamase genes and their cooccurrence in Indian Enterobacteriaceae.Methods:52 third generation cephalosporin resistant isolates were phenotypically detected by combination disk method and screened by PCR to identify class A and class C typeβ-lactamase genes.Results:Of the 52 isolates,94.2%（49） were found harboring any of the bla?,blaCTX-M,blaSHV and blaTEM were present in 82.6%（43/52）, 59.6%（31/52）,and 42.3%（22/52） isolates,respectively.Of the 49 ESBL positive isolates 57.1% （28/49） showed co-occurrence of blaampCwith bla?.On the contrary,the collection from 2009 showed their co-occurrence in 81.4%isolates.Conclusions:The comparative study shows a downward trend for co-existence of bla? with blaampC from 2009 to 2010.Further large scale studies are needed to address the co-occurrence of class A and class Cβ-lactamases in India and the resistance trend occurring over a period of time.

Antimicrobial susceptibility of strains of Enterobacteriaceae isolated from bloodstream infections using current CLSI and EUCAST breakpoints

Understanding local susceptibility patterns is important when selecting antimicrobials for initial empirical antibiotic-therapy of bloodstream infections. Because the determination of susceptibility is dependent on the breakpoints used, the aim of the study was to compare the antimicrobial susceptibility results to different classes of antibiotics of 512 strains of Enterobacteriaceae (200 ESβL positive) isolated from bloodstream using CLSI 2013 and current EUCAST 2013 guidelines to evaluate the impact of break-point discrepancies. The results of the study showed that statistically significant discrepancies (p ≤ 0.001) were found for amoxicillin/clavulanic acid, piperacillin alone or with tazobac-tam, imipenem, meropenem, cefepime (only ESβL negative isolates), amikacin and gentamicin using current CLSI or EUCAST interpretive criteria. Further harmonization of CLSI and EUCAST breakpoints is warranted. This study could give useful information to physicians for managing bloodstream infections caused by Enterobacteriaceae.

Genotypic Diversity and Characterization of Quinolone Resistant Determinants from Enterobacteriaceae in Yaoundé, Cameroon

Background: Enterobacteriaceae causes many types of infections which are often treated with quinolones and fluoroquinolone (Q/FQ). The resistance mechanisms to Q/FQ are usually associated with mutations in the quinolone resistance determining region which alter the conformation of target amino acid residues within the protein and in the qnr genes. This study aimed at determining the antimicrobial resistant profile of a sample of Enterobacteriaceae from Cameroon and the genetic diversity in quinolone-resistant isolates in view of implementing a better management, treatment, control and prevention of the transmission of these resistant strains. Methods: Identification and antimicrobial susceptibility testing was done using VITEK 2. The detection of plamid-mediated quinolone resistance (PMQR) genes was carried out using the conventional PCR method. Sequencing was done using the Applied Biosystem 3500 genetic analyser. DNA fingerprint was obtained using Pulsed-Field Gel electrophoresis. Results: Among 440 Enterobacteriaceae, the most prevalent genera were: Escherichia 178/440 (39.5%);Klebsiella 148/440 (33.6%);Enterobacter 35/440 (8%);Pantoea 28/440 (6.4%);Proteus 14/440 (3.2%) Salmonella 13/440 (3%). Ampicillin resistance showed the highest prevalence with 371/440 (81%) and Imipenem the lowest resistance 9/440 (2.1%). The ciprofloxacin resistance rate was 161/440 (36.6%). The detected plasmid mediated quinolone resistance (PMQR) genes were: qnrA, 2/161 (1.2%);qnrB, 31/161 (19.3%);qnrS, 13/161 (8.1%): Aac (6')Ib-cr, 84/161 (52.2%) and qepA, 3/161 (1.9%). There were several mutations in the parC gene of Klebsiella leading to S80D and S80N substitutions. Two pairs of Klebsiella peumoniae strains were phenotypically and genotypically identical with 100% similarity in the dendrogramme. Conclusion: This study showed that quinolone resistance was high. The PMQR genes contributing to this resistance were diverse. This high PMQR indicates that there has been an unknown circulation of these genes in our community. To avoid the rapid dissemination of these PMQR genes continuous surveillance of antimicrobial resistance should be carried out not only in humans but also in animals to monitor the evolution of these genes.

Occurrence of Extended-Spectrum β-1actamase Producing Enterobacteriaceae （ESBLPE） among Primary School Pupil in Obafemi-Owode, Nigeria

Occurrence of extended-β-1actamase producing enterobacteriaceae （ESBLPE）, which has reduced the antibacterial efficacy and potency of many 3rd generation cephalosporins, was investigated among the primary school pupils. 88 primary school pupils in Obafemi-Owode Local Government, Southwestern Nigeria, including 49 males （55.7%） and 39 females （44,3%） （mean age 12 ± 3） were screened for ESBLPE isolates with exclusion criterion of antimicrobial use in the preceding 2 weeks either as therapy for gastro-intestinal complication or prophylaxis. ESBLPE detected include 4.5% of Eschericia coli, 2.3% of Enterobacter cloaca, 0% Proteus mirabilis, 2.3% Pseudomonas aeruginosa, 1.1% Staphylococcus aureus and 4.5% of Klebsiella oxytoca. 10 （76.9%） of ESBLPE isolates were resistant to disc of cefuroxime （30 μg）, 8 （61.5%） susceptible to amoxicillin/clavulanic （20/10 lag） and low susceptibility of 7 （53.8%） was recorded for ceftazidime （30 lag）. 0% susceptibility was recorded for the ESBLPE isolates to cefuroxime MIC 〉 8 gg/mL and ampicillin MIC 〉 8 lag/mL while E, coli and E. cloca each show 50.0% and P. aeruginosa and K. oxytoca show 100.0% and 75.0% susceptibility to augmentin （MIC 〈 8）. This study has shown a 14.7% proportion of the pupil to harbour ESBLPE from enteric source with increased resistant to most new generation cefuroximes. Therefore, transfer of virulent and antibiotic resistant ESBLPE could be aided by sharing feeding materials while fecal-oral route of transmission cannot be ruled out as hygiene level is very low thereby increasing emergence of virulent resistant enteric strains leading to treatment failure.

Erratum to “Isolation, Characterization and Antimicrobial Resistance Patterns of Lactose-Fermenter Enterobacteriaceae Isolates from Clinical and Environmental Samples” [Open Journal of Medical Microbiology 5 (2015) 169-176]

The lactose-fermenter Enterobacteriaceae are the most frequent cause of clinical infection in our country. The objective of this study was to isolate and identify the most common lactose-fermenter Enterobacteriaceae from clinical samples, including urine, blood, wounds, and sputum, obtained from the local hospital and from environmental samples from a chicken farm, agriculture soil, and water from the Tigris River in Baghdad City. The study also aimed at establishing the antibiotic resistance patterns of the isolated bacteria. A total of 155 bacterial isolates were identified from 10 genera according to the Vitek 2 system. The most common bacterial isolates from the clinical and environmental samples were Escherichia coli and Klebsiella pneumoniae, respectively. The antibiotic resistance patterns showed that all clinical and environmental isolates were multidrug resistant to β-lactam (except carbapenems) drug and aminoglycosides and more sensitive to carbapenems.

Analysis of Enterobacteriaceae Producing Broad-Spectrum Beta-Lactamases in the Intensive Care Unit Setting

Strains of the Enterobacteriaceae family producing ESBL and AmpC broad-spectrum beta-lactamases that may survive in the hospital setting potentially cause infection in hospitalized patients due to contaminated objects or health care workers’ hands. Over a period of two months (November-December 2010), a single epidemiological study of microbial contamination of air, surfaces and health care workers (swabs from both nostrils and the right hand without a glove) was carried out at two intensive care units of the University Hospital Olomouc, Czech Republic. The bacteria were identified using standard microbiological methods. Phenotypic detection of ESBL and AmpC enzymes and basic genetic analysis of ESBL- and AmpC-positive isolates was performed. The same approach was used to identify and analyze bacteria isolated from clinical samples of patients hospitalized at the above departments over the study period. From a total of 140 environmental samples collected over the study period, 21 isolates of the Enterobacteriaceae family were identified, with ESBL and AmpC production being detected in 4 and 7 isolates, respectively. Among patients’ clinical samples, 10 ESBL- and 6 AmpC-positive isolates were detected. No similarity was found between environmental isolates and strains isolated from patients.

Occurrence of Enterobacter sakazakii(Cronobacter)and Other Enterobacteriaceae in Commercial Powdered Infant Formula in Abidjan,Ivory Coast

To determine the occurrence of Enterobacter sakazakii and other Enerobateriaceae in commercial powdered infant formula (PIF), 185 packages of PIF from different manufacturers, supermarkets and drug-stores in Abidjan were analyzed. Ten g of sample was homogenized in 90 ml of buffered peptone water (PBW, Biorad, Paris) for further studies. Enterobacteriaceae (coliforms) were enumerated according to French Association of Standardization methods. E. sakazakii was detected according to Kandhai’s method. Bacteria were identified using API20 system. Thirty-eight samples (20.5%) were positive for Enterobacteriaceae. Twenty-four samples (13%) yielded Enterobacter sakazakii. Other Enterobacteriaceae isolated included Pantoea spp. 21 (11.5%), Klebsiella pneumoniae subsp. Pneumonia 8 (4.3%), Citrobacter diversus 1 (0.5%), Citrobacter freundii 1 (0.5%), Enterobacter cloacae 1 (0.5%), Salmonella reading 1 (0.5%), Serratia ficara 1 (0.5%) Serratia odorifera 1 (0.5%). This study is the first report to describe the contamination of PIF from Abidjan with E. sakazakii and several other Enterobacteriaceae that could be opportunistic pathogens. Therefore, well-controlled studies need to be conducted to assess the extent of risk associated with contaminated PIF for infants in Abidjan.