In vitro efficacy of Daptomycin against clinical isolates of Methicillin-resistant Staphylococcus aureus (MRSA).

This descriptive cross-sectional study was performed in the Department of Microbiology, Fauji Foundation Hospital, Rawalpindi, from March 2019 to September 2019 to determine the in vitro efficacy of Daptomycin against clinical isolates of Methicillin-Resistant Staphylococcus aureus (MRSA). Consecutive non-probability sampling technique was used and a total number of 270 patients' Pan Cultures having MRSA growth on Cefoxatin Disc with size less than 22 mm zone size were included in the study. Cultures were inoculated on MacConkey, Chocolate and Blood agar and then incubated for 24 hours at 37 degree Celsius. After incubation, Coagulase test, Catalase test and Gram staining technique were used for further identification. Minimum Inhibitory Concentration (MIC) of the isolates for Daptomycin was obtained by using E strips (Oxoid UK) according to Clinical & Laboratory Standards Institute (CLSI) guidelines. The mean age of the patients was 46.73±12.22 years, and the study included 147 (54.44%) males and 123 (45.56%) females. Regarding the type of specimen, there were 154 (57.04%) pus specimens, 54 (20.00%) blood specimens, 27 (10.00%) fluid specimens, 18 (6.67%) urine specimens, 10 (3.70%) high vaginal swabs (HVS) specimens and 7 (2.59%) sputum specimens. Daptomycin was effective in 264 (97.78%) patients with MIC range from .015 to 1 µg/ml on E strip.

Phenotypic detection of extended-spectrum beta-lactamase in multidrug-resistant acinetobacter baumannii isolated in Fauji Foundation Hospital Rawalpindi.

OBJECTIVE: To evaluate the phenotypic detection of extended-spectrum betalactamase in multidrug-resistant acinetobacter baumannii. METHODS: The cross-sectional study was conducted at the Department of Microbiology, Fauji Foundation Hospital, Rawalpindi, Pakistan, from August 2018 to April 2019, after the ethical approval from the Institutional Review Committee. Consecutive Non- probability sampling technique was used, and comprised clinical specimens, including pus, blood, sputum, urine, tracheal tubes and canula double lumen, which were processed using standard protocols. Colonies of acinetobacter baumannii were identified by gram staining and Analytical Profile Index-20E kit. Combination disc method was used for the identification of extended-spectrum beta-lactamse. Clinical and Laboratory Standards Institute guidelines were used for antimicrobial susceptibility. Data was analysed using SPSS 22 and Sample size was calculated by using earlier study with 5 % margin of error and 95 % confidence level. RESULTS: Of the 78 isolates, 58(74.4%) related to females and 20(25.6%) to males. There was no extended-spectrum beta-lactamse producer. Imipenem, meropenem, cefotaxime, ampicillin and ceftazidime showed 100% resistance, while colistin and polymyxin B were sensitive to all strains. The incidence rate was high in samples isolated from tracheal tubes 47(60.3%), followed by pus 21(26.9%). Age was not found to be a significant factor (p>0.05). CONCLUSIONS: Acinetobacter baumannii showed a high resistance to multiple drugs and was not confined to any specific age group. Colistin and polymyxin B were found to be better choices.

Endogenous methicillin resistant staphylococcus aureus as a source of post-operative surgical site infections.

OBJECTIVE: To determine frequency of Endogenous Methicillin Resistant Staphylococcus aureus (MRSA) in pre-operative patients and its frequency in Surgical Site Infections (SSIs) post operatively. METHODS: It was a descriptive cross sectional conducted at Department of Microbiology, Fauji Foundation Hospital Rawalpindi (FFH), Pakistan. Samples were collected from 1st November-31st May 2018. Total 75 samples were collected during the period. Consecutive non-probability sampling technique was utilized. Specimens were collected from nose, axilla and groin of preoperative patients. Methicillin Resistant Staphylococcus aureus was identified if only isolated from these sites. Patients were followed till his/her discharge from the hospital and if they developed infection post operatively, pus specimen from infected site was also collected and identified. RESULTS: Out of 75 specimen preoperatively, 11(14.7%) were identified as endogenous MRSA. From these, 33(44%) developed Surgical site infections (SSIs), among them 19(57%) were MRSA (09 endogenous, 10 exogenous), 7(21.2%) were Escherichia coli, 3(9.1%) were Klebsiella pneumoniae, 3(9.1%) were Enterococcus faecalis and 1(3%) was Methicillin Sensitive Staphylococcus aureus. CONCLUSIONS: The results of this study determined that Endogenous Methicillin Resistant Staphylococcus aureus (MRSA) could be isolated from patients going for surgery if microbiological screening was done at the time of admission. This could prevent patients from Surgical Site Infection Post operatively by these endogenous MRSA. This search and wipe out strategy is able to curtail the events of outbreak, reduce hospital stay and decrease budget of the hospital by providing guidance in choice of empirical therapy for infection.

Antimicrobial susceptibility pattern of Methicillin-resistant Staphylococcus Aureus isolates in Fauji Foundation Hospital Rawalpindi.

The current study was conducted in the Department of Microbiology, Fauji Foundation Hospital, Rawalpindi, from July 2018 to January 2019. The main purpose of the study was to evaluate Methicillin-resistant Staphylococcus aureus antimicrobial susceptibility pattern. Clinical samples were collected and cultured according to Clinical and Laboratory Standards Institute (CLSI) guidelines. A total of 90(30%) samples were found to be methicillin-resistant out of 300 samples of Staphylococcus aureus. Major isolates were 42 (46.67%) from pus and 22 (24.44%) from tracheal tubes. The incidence ratio of Methicillin-resistant Staphylococcus aureus was high in the samples isolated from 69 (76.67%) females compared to those of 21 (23.33%) males. Patients were more in the age group of 41 to 60 years. Vancomycin 90 (100%) was sensitive to all strains followed by Chloramphenicol 66 (73.33%) and Doxycycline 52 (57.78%). Imipenem, Meropenem andAugmentin showed resistance to all strains.

Susceptibility pattern of tracheal tube isolates from Intensive Care Unit of Fauji Foundation Hospital Rawalpindi.

OBJECTIVE: To determine the prevalence of resistant pathogens and their antimicrobial susceptibility pattern in an intensive care unit. METHODS: The cross-sectional observational study was conducted at Foundation Hospital, Rawalpindi, Pakistan, from May to September 2016, and comprised tracheal tubes which were collected in sputum culture bottles from patients with clinical findings of ventilator associated pneumonia. The tubes were cultured to locate the resistant pathogens. RESULTS: A total of 113 different strains of bacteria were isolated from 80 patients. The main isolated bacteria was acinetobacter baumannii 45(39.8%) followed by klebsiella pneumonia 14(12.3%) and methicillin-resistant staphylococcus aureus 13(11.5%). Polymyxin B was the most appropriate drug for treating patients infected with acinetobacter baumannii with a sensitivity of 64% while vancomycin and linez oli dhad 100% sensitivity for methicill in - resistant staphylococcusaureus. CONCLUSIONS: Acinetobacter baumannii was the most prevalent strain in tracheal tubes and polymyxin B was the most effective medicine.

Mycobacterium tuberculosis infection and resistance to rifampicin with GeneXpert®MTB/RIF: a single-center experience on bronchoalveolar lavage samples in renal failure patients.

Patients with end-stage renal disease (ESRD) are immunocompromised and are more at risk to develop and acquire Mycobacterium tuberculosis (MTB) infection. However, risk assessment is uncertain. The objective of current research was to study the frequency of MTB infection in ESRD patients . For this purpose, bronchoalveolar lavage (BAL) samples were evaluated for the presence of MTB by using GeneXpert®MTB/RIF test. We analysed 350 clinical samples of BAL collected from a tertiary care hospital in Pakistan, from September, 2015 to July, 2016. We performed the GeneXpert®test on each sample. According to our results prevalence of MTB was observed in 1.7% of bronchoalveolar lavage (BAL) samples taken from patients with chronic kidney diseases. All the positive samples were susceptible to rifampicin. There is a low prevalence of MTB infec tion (pulmonar y tuberculosis) in patients with chronic kidney disease in our setup. Suspected patients can be diagnosed by using GeneXpert®MTB/RIF testing on bronchoalveolar lavage samples.

Hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA) from Pakistan: molecular characterisation by microarray technology.

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Pakistan is known to be high, but very few studies have described the molecular epidemiology of the different MRSA clones circulating in the country. Forty-four MRSA isolates were collected from two tertiary care hospitals of the Rawalpindi district of Pakistan. All strains were identified by a conventional phenotypic method and then subjected to genotyping by microarray hybridisation. Six clonal complexes (CCs) and 19 strains were identified. The most commonly identified strains were: (i) Panton-Valentine leucocidin (PVL)-positive CC772-MRSA-V, "Bengal Bay Clone" (ten isolates; 22.3%), (ii) ST239-MRSA [III + ccrC] (five isolates) and (iii) a CC8-MRSA-IV strain, as well as CC6-MRSA-IV (both with four isolates; 9.1% each). Several of the strains detected indicated epidemiological links to the Middle Eastern/Arabian Gulf region. Further studies are needed to type MRSA from countries with less known epidemiology and to monitor the distribution and spread of strains, as well as possible links to global travel, migration and commerce.

Detection and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus from Table Eggs in Haripur, Pakistan.

Table eggs are nutritionally important food consumed globally. Despite being protected inside the hard shell and a semipermeable membrane, the egg contents may be contaminated with microbes and thus become a possible carrier of infectious agents to humans. A number of medically significant bacterial species such as Salmonella enterica, Listeria monocytogenes, and Yersinia enterocolitica have already been reported from table eggs. More important is the presence of antimicrobial-resistant bacterial strains in this food source. The present study was aimed at detection and characterization of Staphylococcus aureus from table eggs collected from different retail shops in Haripur city of Pakistan. Staphylococci were isolated from 300 eggs collected from December 2015 to May 2016. S. aureus isolates were tested for antimicrobial susceptibility using broth microdilution and characterized using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCCmec) typing, and spa typing. The presence of Panton-Valentine leukocidin and antimicrobial resistance genes were detected using PCR. Staphylococci were isolated from 21.3% (64/300) of the table eggs tested. Of those, 59% (38/64) were identified as S. aureus, of which 33 (86.8%) were positive for mecA (MRSA, methicillin-resistant S. aureus). All MRSA were multidrug resistant (resistant to two or more antimicrobial classes), contained aac-aph (encoding aminoglycosides), and were pvl+. Using MLST, spa typing, and SCCmec typing, three genotypic patterns were assigned: ST8-t8645-MRSA-IV, associated with USA300; and ST772-t657-MRSA-IV and ST772-t8645-MRSA-IV, both characteristic of the Bengal Bay community-associated MRSA clone. Molecular typing by PFGE revealed that the bacterial population was highly homogenous with only two patterns observed. This study is the first report of detection of human-associated pvl+ MRSA from table eggs. The genetic similarities of MRSA present in the eggs to that of humans may suggest human to poultry transmission of MRSA via contamination.

Multidrug resistance in Gram-negative pathogens isolated from patients with chronic kidney diseases and renal transplant.

Multidrug resistance (MDR) in gram-negative pathogens is the emerging threat to clinicians. The current study was designed to determine the prevalence and pattern of multidrug resistance in gram-negative clinical isolates. It was conducted at the COMSATS Institute of Information Technology, Islamabad, Pakistan, from June to October 2014. Of the 8, 300 samples collected, 729(8.8%) clinically important gram-negative pathogens were retrieved. These pathogens were subjected to phenotypic and biochemical detection and were further processed for multidrug resistance pattern. It was observed that gram-negative pathogens were simultaneously resistant to many antibiotics. The prevalence of extended spectrum b-lactamase phenomenon was 220(100%) in Klebsiella pneumoniae, 195(75%) in Escherichia coli. Resistance to carbapenem was 174(79%) in Klebsiella pneumoniae and 14(5.4%) in Escherichia coli. Resistance against fluoroquinolones also displayed an escalating trend. The current study found that resistance against antibiotics was displaying a drastic increase in chronic renal patients.

Candidiasis: Prevalence and resistance profiling in a tertiary care hospital of Pakistan.

OBJECTIVE: To determine Candida colonisation/infection in renal transplant patients and to determine the resistance pattern against antifungal drugs. METHODS: This prospective, observational study was conducted at Al-Sayyed Hospital, Rawalpindi, Pakistan, from January to October 2014, in collaboration with the Microbiology and Public Health Laboratory's, Islamabad campus..The clinical specimens investigated included respiratory tract secretions, blood, urine, high vaginal swab, skin scrapings, and plastic devices samples. RESULTS: Of the 7,850 samples, 164(2.08%) were positive for Candida. Candida albicans were most prevalent as they were found in 114(69%) samples. Besides, 56(34%) of the positive samples were resistant to one or more antifungal agents. Highest resistance was obtained against fluconazole. We found only 5(3.04%) positive samples of Candida glabrata; of them, 3(60%)were resistant. In case of Candida spp, 27(48%) resistance was observed. In Candida albicans, 23(41%) of the samples were found to be resistant. Most of the Candida isolates was recovered from bronchial alveolar lavage. CONCLUSIONS: Although Candida albicans remained the main responsible species for Candida infections, but non-albican Candida species are also emerging.