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.

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.

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.

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.

Insights into the microbiological and virulence characteristics of bacteria in orthopaedic implant infections: A study from Pakistan.

The exponential increase in the prevalence of multidrug resistant bacteria has resulted in limiting surgical treatment options globally, potentially causing biofilm-related complications, implant failure, and severe consequences. This study aims to isolate and characterize bacteria from post-surgical orthopaedic implant infections and screening for multiple antibiotic resistance. A cross-sectional study was conducted, involving isolation of forty-four dominant pathogenic bacterial isolates from 16 infected implant samples from across Islamabad and Rawalpindi. Out of forty-four, 38% cocci and 61% bacilli were obtained. Approximately 90% of isolates showed multiple antibiotic resistance (MAR) index of more than 0.2. Eleven strains were identified via 16S rRNA gene sequencing as Pseudomonas aeruginosa, Bacillus spp., Planococcus chinensis, Staphylococcus, Escherichia coli and Enterobacter cloacae. The bacterial strain E. coli MB641 showed sensitivity to Polymyxin only, and was resistant to all other antibiotics used. Maximum biofilm forming ability 0.532 ± 0.06, 0.55 ± 0.01 and 0.557 ± 0.07 was observed in Pseudomonas aeruginosa MB663, Pseudomonas aeruginosa MB664 and Bacillus spp. MB647 respectively after 24 hours of incubation. EPS production of bacterial strains was assessed, the polysaccharides and protein content of EPS were found to be in the range of 11-32 µg/ml and 2-10 µg/ml, respectively. Fourier transform infrared spectroscopic analysis of EPS showed the presence of carbohydrates, proteins, alkyl halides, and nucleic acids. X-ray diffraction analysis revealed crystalline structure of EPS extracted from biofilm forming bacteria. These findings suggest a high prevalence of antibiotic-resistant bacteria in orthopaedic implant-associated surgeries, highlighting the urgent need for ongoing monitoring and microorganism testing in infected implants.

Risk Factors for Acute Brucellosis in Patients on the Day of Admission at Selected Hospitals of Abbottabad, Pakistan.

Brucellosis is a neglected zoonotic disease of ruminants. It causes severe health problems in humans and significant economic loss. Only a limited number of studies have been conducted in Pakistan to determine the prevalence of human brucellosis and related risk factors. The objectives of the current cross-sectional study were to determine the prevalence of anti-Brucella antibodies in sera collected from symptomatic patients at three hospitals of Abbottabad using a commercial slide agglutination test (SAT) and to determine risk factors for brucellosis for these patients. Five hundred blood samples were collected. A questionnaire was filled in for each patient to obtain information on age, gender, living area, brucellosis associated symptoms, associated risk factors, pregnancy and abortion history. A total of 13.6% (n = 68) patients were found to be SAT positive and in 83.3% (n = 57) of these samples Brucella DNA was detected by genus specific RT-PCR for BCSP-31 gene. Statistical analysis was performed to determine odd ratios, risk ratios, 95% confidence intervals, and p-values. The prevalence of brucellosis by SAT was reported to be higher in women (14.6%, n = 44) than in men (12.1%, n = 24). The age group 25-50 years was found to be at higher risk for brucellosis (14.5%, n = 50) "animal contact" was reported as the main risk factor followed by "consumption of raw animal products." Out of 131 pregnant women and 21 patients had abortion, the seropositivity of Brucellosis was 9.9% and 23.8%, respectively. The present study reports a striking prevalence of brucellosis among patients including pregnant women at three hospitals of Abbottabad. These findings may foster strategies for controlling human brucellosis at household level, raising of awareness about brucellosis in hospital and family doctors, and finally in setting up an eradication program in the dairy industry.