pKpQIL-like plasmid contributes to the dissemination of blaNDM-1 and plasmid mediated quinolone resistance determinants among multi drug resistant Klebsiella pneumoniae in Assiut university hospital, Egypt.

Background and Objectives: Concomitant carriage of blaNDM-1 and plasmid mediated quinolone resistance determinants (PMQRs) by multi drug resistant (MDR) Klebsiella pneumoniae (K. pneumoniae) has increased globally, often related to their presence on transmissible plasmids. In this study, we hypothesized the presence of blaNDM-1 and PMQRs on a single conjugative plasmid that circulates among K. pneumoniae strains isolated from Assiut University Hospital. Materials and Methods: Twenty-two clinical MDR K. pneumoniae strains harboring both blaNDM-1 and PMQRs were genotyped using pulsed field gel electrophoresis. Horizontal transfer of blaNDM-1 and PMQRs was evaluated by conjugation and trans-conjugants were screened for the presence of both genes and integron by PCR. Trans-conjugant's plasmid DNA bands were purified using agarose gel electrophoresis and different DNA bands were screened for blaNDM-1 and PMQRs. Plasmids carrying blaNDM-1 and PMQRs were typed by PCR based replicon typing. Results: All MDR K. pneumoniae contained class 1 integron and belonged to 15 pulsotypes. BlaNDM-1 and PMQRs were co-transferred in each conjugation process. Multiple replicons (5-9 types) were detected in each trans-conjugant; with IncFIIK and IncFIB-KQ replicons being common among all trans-conjugants. Both blaNDM-1 and PMQRs were detected on a pKpQIL-like multi-replicon plasmid that was present in all K. pneumoniae strains. Conclusion: In view of these results, the presence of blaNDM-1 and PMQRs on pKpQIL-like plasmid that existed in multiple unrelated K. pneumoniae isolates is highly suggestive of the circulation of pKpQIL-like MDR plasmids in our hospitals. Moreover, carriage of integrons by the-circulating MDR plasmids increases the risk of dissemination of antimicrobial resistance among pathogens.

Modified PFGE protocol for improving typeability of DNA degradation susceptible nosocomial Klebsiella pneumoniae.

Introduction. PFGE is the 'gold standard' method for bacterial subtyping. However, many strains are non-typable by this approach because of DNA degradation by nucleases action.Aim. To evaluate a modified PFGE protocol for typing nosocomial isolates of Klebsiella pneumoniae.Methods. Twenty- five K. pneumoniae isolates previously exposed to DNA degradation were used to optimize an extraction method for elimination of DNases activity before applying Xba1 enzyme. Introducing of sodium dodecyl sulfate (SDS) in different concentrations to the extraction buffer was evaluated for protecting genomic DNA molecule from degradation by nucleases.Results. Addition of 3 % SDS in combination with 3 % N-lauryl sarcosine to the extraction buffer was found to reduce the previously experienced nuclease activity. Pre-examination of plug quality prior to the digestion phase could efficiently reduce the expense of the wasted enzyme.Conclusion. We have successfully devised a PFGE protocol that enhanced the typeability of nosocomial K. pneumoniae.

Seropositivity in animals and risk of occupational brucellosis among abattoirs personnel associated with poor work practices and absence of safety policy in Egypt.

Background Occupational hazards are the leading cause of morbidity and mortality among abattoirs personnel and animal workers. These hazards result from direct or indirect exposure to potential infection and several distressing events during routine procedures. Objectives To serologically investigate the potential occupational brucellosis hazard at Egyptian abattoirs. To provide an insight on the needed biosafety practices that should be implemented to mitigate the spread of occupational brucellosis among abattoir workers. Methods Two hundred and thirty (n = 230) blood samples were collected from animals in two Egyptian abattoirs. The rose Bengal test was used to evaluate the seroprevalence of Brucella in abattoir animals. A questionnaire was distributed among abattoir personnel to address biosafety gaps and deficiencies as a cause of occupational brucellosis. Results The overall seroprevalence of Brucella using the rose Bengal test was 75.2% in the two targeted abattoirs. It was obvious that there are gaps of malpractices and inconvenient behavior among individuals of the targeted community. Conclusions The current findings reveal the missing role of concerned authorities and lack of written safety policy. The data highlights the need for further research, including isolation and characterization of the causative agents, and reliable epidemiological studies.

Epidemiological typing of multidrug-resistant Klebsiella pneumoniae, which causes paediatric ventilator-associated pneumonia in Egypt.

PURPOSE: Multidrug-resistant Klebsiella pneumoniae is a common nosocomial pathogen that plays an important role in ventilator-associated pneumonia (VAP). This study aimed to define the clonal relatedness of K. pneumoniae strains isolated from paediatric VAP in addition to those isolated from environmental samples. METHODOLOGY: This study included 19 clinical and 4 environmental K. pneumoniae isolates recovered from the paediatric intensive care unit (PICU) in Assiut University Children's Hospital. The K. pneumoniae isolates were confirmed by biotyping using API strips and subjected to antimicrobial susceptibility testing. The genes coding K1 and K2 capsular types were detected by PCR. The clonal relationships between the K. pneumoniae isolates were determined by pulsed-field gel electrophoresis (PFGE). RESULTS: Ten resistotypes were detected among all the K. pneumoniae isolates, while PFGE identified seventeen K. pneumoniae pulsotypes. Similar PFGE patterns were found between environmental and clinical isolates and between isolates recovered from different patients, suggesting the circulation of K. pneumoniae pathogens in the PICU and the role of the environment in the spread of infection. No correlation was found between the resistotypes and pulsotypes of the K. pneumoniae isolates. PFGE showed higher discriminatory power for the typing of nosocomial K. pneumoniae [Simpson's diversity index (DI)=0.96] than resistotyping (DI=0.72). CONCLUSION: As far as we know, this is the first report of the isolation of the same multidrug-resistant (MDR) K. pneumoniae pulsotype from patients and environmental samples in the same hospital ward in Egypt. This study provides a step on the way to understanding the genotyping and epidemiology of MDR K. pneumoniae for enhanced prevention of bacterial transmission.