Bacterial contamination of Pakistani currency notes from hospital and community sources

Objective: We determined the bacterial contamination and antibiotic resistance profile of circulating Pakistani currency notes collected from hospital and community sources

Methods: This prospective study was organized from July to December 2015 in the Microbiology Department of The Children's Hospital and The Institute of Child Health Lahore. It was done on one hundred currency notes of four different denominations collected from various groups of people in sterile polythene bags. Gram staining, colony morphology and various biochemical tests were used to identify the bacterial isolates. Kirby Bauer disc diffusion method was used to observe the antibacterial drug resistance

Results: There were 11 different types of bacterial species which contaminated 97 [97 percent] currency notes. The bacterial isolates discovered from paper currency notes included Klebsiella spp. [26.0 percent], Coagulase-negative Staphylococci [CoNS] [18.3 percent], E. coli[14.5 percent], Pseudomonas spp. [13.7 percent], Citrobacter spp. [11.5 percent], Enterobacter spp. [5.3 percent], Acinetobacter spp. [5.3 percent], Streptococcus spp. [2.3 percent], Shigella spp. [1.5 percent], Salmonella spp. [0.8 percent] and Pantoea spp. [0.8 percent]. Most of the Gram-positive isolates were resistant to penicillin and ampicillin. None of the Gram-negative isolates found to be resistant to amikacin, cefoperazone-sulbactam and piperacillin-tazobactam

Conclusion: The currency notes circulating in hospital and community are contaminated with highly pathogenic and some multi-drug resistant bacteria. These currency notes could be a potential source of nosocomial and community-acquired infections

Antibacterial activity of penicillins alone and in combination with different agents against Staphylococcus aureus

Difficulties in the treatment of the resistant strains of Staphylococcus aureus, which is a frequent cause of nosocomial infections in paediatric patients, has prompted this research to empower the usage of various combinations of penicillin. During the study period 17,452 clinical samples were processed for culture. The positive cultures yielded 564 strains of S. aureus. Out of these, 362 [64.2%] isolates were found to be methicillin sensitive S. aureus [MSSA] and 202 [35.8%] methicillin resistant S. aureus [MRSA]. The frequency of S. aureus isolates from male patients [355; 63.1%] was found to be higher than female patients [209; 36.9%] and those from indoor wards [441; 78.2%] were more than the outdoor wards [123; 21.8%]. Frequency distribution of S. aureus showed to be highest among blood 342 [60.6%] and cerebrospinal fluid 100 [17.8%] samples. The sensitivity pattern of MSSA with piperacillin-tazobactam was 344 [95.0%], ampicillin-sulbactam 340 [93.9%], co-amoxiclav 332 [91.8%] and ampicillin-oxacillin 257 [71.0%]. MRSA susceptibility to piperacillin-tazobactam was 143 [71.0%], ampicillin-sulbactam 114 [56.6%], co-amoxiclav 61 [30.2%] and ampicillin-cloxacillin 18 [9%]. The Cochran Mantel Haenszel test showed that the effectiveness for each penicillin was associated significantly [p<0.05] with both the MSSA and MRSA. The combinations of piperacillin-tazobactam, ampicillin-sulbactam, co-amoxiclav and ampicillin-cloxacillin exhibited higher efficacy than using them alone to combat Staphylococcal infections

Acute bacterial meningitis in children presenting to the children's hospital Lahore before and after pneumococcal vaccine in Pakistan national immunization program; a comparison

Objective: To describe bacteriological profile, morbidity and mortality of acute bacterial meningitis [ABM] in children and to compare these parameters before and after the introduction of Pneumococcal vaccine in Pakistan National Immunization Program

Methods: The present descriptive study was conducted at the Department of Paediatric Medicine of The Children's Hospital Lahore from January 2012 to December 2015. A total of 503 children one month to five years of age admitted with diagnosis of meningitis were included. Complete blood count, CSF cytology, biochemistry, culture sensitivity and blood culture sensitivity were performed

Results: Frequency of meningitis decreased by 50% in 2013-2015 [199 [2012] vs 304 [2013-2015]. Most children in both groups were under one year of age. More neurological complications were seen in the group 2, 20% vs 17%. CSF culture positivity decreased from 12% to 6.6%. Streptococcus pneumoniae isolation decreased from 5 [2.5%] in 2012 to 4 [1.3%] in 2013-2015. Refusal to take feed [p=0.002], impaired sensorium [p=<0.001], severe malnutrition [p=0.001], prolonged duration of symptoms [p=<0.001] and incomplete vaccination status [0.005] were associated with mortality. Mortality rate decreased from 20 [10%] in 2012 to 17 [5.6%] in 2013-2015 but more children developed neurological sequelae 2.7% versus 1%

Conclusion: Acute bacterial meningitis mostly affected children <1 year. Frequency of Streptococcus pneumoniae and mortality of meningitis decreased significantly after PCV but more neurological complications developed in those children who were unvaccinated in 2013-2015 compared to 2012

Phenotypic characterization of extended-spectrum-beta-lactamase producing E. from healthy individuals, patients, sewage sludge, cattle, chickens and raw meat

Objective: The present study aimed to determine the frequency and antimicrobial profile of ESBL-producing isolates of f. coli in different environments

Methods: This cross-sectional study was conducted at The Children's Hospital and The Institute of Child Health, Lahore from July to December 2015. The faecal specimens from healthy individuals, patients, sewage sludge, cattle, chickens and raw meat [n = 122] were processed for microbiological analysis using MacConkey agar supplemented with cefotaxime. The identification of organisms was confirmed by API 10S and antimicrobial resistance profile was recorded by Kirby-Bauer disc diffusion method

Results: On the basis of screening, 77 [63.0%] specimens were found to be positive for ESBL production

The confirmation of 74 [60.0%] ESBL producing f. coli was done using double disc synergy test [DDST]

The frequency of ESBL producing E. coli was found to be 17 [57.0%] in healthy individuals, 15 [53.0%] in patients, 10 [66.0%] in cattle faeces, 5 [71.0%] in sewage sludge, 14 [70.0%] in raw meat and 13 [59.0%] in chicken faeces. All of these isolates were resistant to cephalosporins and some of these were resistant to fluoroquinolones and meropenem. None of the isolates showed resistance to cefoperazone-sulbactam, imipenem, piperacillin-tazobactam and amikacin

Conclusion: The prevalence of ESBL-producing E. coli was recorded in all the environments, suggesting a global expansion of these enzymes

Detection of AmpC beta-lactamase producing bacteria isolated in neonatal sepsis

Objective: The objective of this study was to determine the occurrence and antimicrobial profile of AmpC beta-lactamase producing bacteria

Methods: The study was conducted at The Children's Hospital and The Institute of Child Health Lahore, Pakistan, during September 2011 to June 2012. A total number of 1,914 blood samples of suspected neonatal septicemia were processed. Isolates were identified using Gram's staining, API 20E and API 20NE tests. Gram negative isolates were screened for AmpC beta-lactamase production against ceftazidime, cefotaxime and cefoxitin resistance and confirmed by inhibitor based method

Results: Total number of 54 [8.49%] Gram positive and 582 [91.5%] Gram negative bacteria were identified. Among Gram negative isolates 141 [22%] were AmpC producers and found to be 100% resistant to co-amoxiclav, cefoxitin, ceftazidime, cefotaxime, cefuroxime, cefixime, ceftriaxone, cefpodoxime, gentamicin, amikacin and aztreonam. Less resistance was observed against cefepime [30.4%], sulbactamcefoperazone [24.8%], piperacillin-tazobactam [10.6%], ciprofloxacin [20.5%] and meropenem [2.1%]. All the isolates were found sensitive to imipenem. The patients harbored AmpC beta-lactamases were on various interventions in which intravenous line was noted among [51.1%], naso-gastric tube [37.6%], ambu bag [8.5%], endotracheal tube [3.5%], ventilator [2.1%] and surgery [0.7%]

Conclusion: Extensive use of invasive procedures and third generation cephalosporins should be restricted to avoid the emergence of AmpC beta-lactamases in neonates

Multidrug resistant AmpC [beta]-lactamase producing Escherichia coli isolated from a paediatric hospital

The objective of the study was to observe the antimicrobial resistance of AmpC [beta]-lactamase producing E. coli. Six hundred and seventy E. coli were isolated from 20,257 various pathological samples collected from The Children's Hospital and Institute of Child Health, Lahore, Pakistan. The isolates showed resistance to ceftazidime which were further examined for AmpC [beta]-lactamase activity by Disc Potentiation method. There were 670 isolates of E. coli out of which 85 [12.6%] were AmpC [beta]-lactamase producers. Risk factors like intravenous line [76.5%], endotracheal tube [22.4%], surgery [12.9%] and urinary catheters [7.1%] were found to be associated with infection caused by AmpC [beta]-lactamase producing E. coli. Antimicrobial resistance pattern revealed that AmpC producing E. coli were highly resistant to coamoxiclav, ceftazidime, cefotaxime, cefuroxime, cefixime, ceftriaxone and cefoxitin [100% each]. Least resistance was observed against sulbactam-cefoperazone [14.1%], cefepime [7.1%], piperacillin-tazobactam [5.9%] and none of the isolates were resistant to imipenem and meropenem. The minimum use of invasive devices and strict antibiotic policies can reduce the spread of AmpC [beta]-lactamase producing E. coli

Detection of extended-spectrum beta -lactamases in Klebsiella pneumoniae: comparison of phenotypic characterization methods

Extended-spectrum beta -lactamase producing K. pneumoniae is a serious threat to the patients. This manuscript shows the comparison of phenotypic characterization methods used for ESBL K. pneumoniae and frequency distribution of these isolates in various clinical samples. Eleven different types of pathological samples collected on various time intervals were analyzed. K. pneumoniae were identified with API 20E system [bioMerieux] and initial screening of ESBL K. pneumoniae was performed using the ceftazidime antimicrobial disc. Double-disc synergy test [DDST] and CLSI confirmatory test were compared for the phenotypic detection of ESBL K. pneumoniae. A total number of 214 ESBL producing K. pneumoniae were isolated from various clinical samples. Frequency distribution of ESBL producing K. pneumoniae was found to be highest among blood 117 [54.7%] and urine 46 [21.5%] samples. Data regarding the use of various interventions among these patients showed most common presence of intravenous line 209 [97.7%] and urinary catheters 46 [21.5%]. Comparison of DDST and CLSI confirmatory test showed that the DDST detected 145 [67.8%] isolates while 213 [99.5%] ESBL K. pneumoniae were characterized by CLSI confirmatory test. The use of CLSI confirmatory test is very efficient in the early detection of ESBL K. pneumoniae especially when the facilities for molecular characterization are not available

Antibiotic therapy in pyogenic meningitis in paediatric patients

To isolate and identify the causative pathogen, antibiotic sensitivity testing and success rate of empirical antibiotic therapy in pyogenic meningitis. Analytical study. The Children's Hospital and Institute of Child Health, Lahore, Pakistan, from March to July 2012. The study was performed on 72 culture positive meningitis cases in children less than 15 years of age. This therapy was evaluated by monitoring the patient's clinical picture for 14 - 21 days. The collected data was analyzed by Chi-square test. Seventeen different bacteria were isolated. The most commonly occurring bacteria were coagulase negative Staphylococci [25%], E.coli [12.5%], Klebsiella pneumoniae [8.3%], Streptococcus pneumoniae [8.3%] and Pseudomonas aeruginosa [8.3%]. All the bacteria were sensitive to vancomycin [96.7%], meropenem [76.7%], amikacin [75%], ciprofloxacin [65.3%], chloramphenicol [46.5%], ceftazidime [44.2%], cefepime [41.9%], co-amoxiclav [38.0%], oxacillin [34.8%], cefotaxime [21.4%], penicillin [20.7%], ceftriaxone [18.6%], cefuroxime [14%] and ampicillin [6.9%]. The combination of sulbactam and cefoperazone showed antimicrobial sensitivity of 81.4%. The success rate of empirical antibiotic therapy was 91.7%. It was found that Gram negative bacteria were the major cause of pyogenic meningitis. Mostly there were resistant strains against all commonly used antibiotics except vancomycin. All empirical antibiotic therapies were found to be most successful

Phenotypic characterization of ESBL producing Enterobacter cloacae among children

The emergence of ESBL producing Enterobacter cloacae in clinical isolates is posing a serious threat for treating nosocomial infections. The aim of the study was to determine the frequency of extended spectrum beta-lactamase [ESBL] producing Enterobacter cloacae and to compare the phenotypic methods used for the characterization of ESBL producing strains. This cross sectional observational study was conducted during April 2011 to March 2012 at Microbiology department of The Children's Hospital and Institute of Child Health, Lahore. A total number of 20,257 various clinical samples were analyzed during the study period. Enterobacter cloacae were identified using API 20E system and ESBL detection was carried out using double-disk synergy test [DDST] and CLSI confirmatory test. Enterobacter cloacae were isolated from 221 samples, out of which 33 [14.93%] were ESBL producers and 188 [85.07%] were non-ESBL producers. The gender distribution of ESBL producing Enterobacter cloacae was 21 [63.6%] in males and 12 [36.4%] in females. Highest frequency [63%] of ESBL producing Enterobacter cloacae was detected in blood samples. Comparison of DDST and CLSI confirmatory test showed that 25 [75.75%] isolates were characterized by DDST and 33 [100%] using CLSI confirmatory test. The present study shows moderately high frequency of ESBL producing Enterobacter cloacae among children. DDST was found to be less efficient in ESBL detection as compared to CLSI confirmatory test

Detection of AmpC beta-lactamase in clinical isolates of Escherichia coli among children

The production of AmpC beta-lactamases is one of the common mechanisms of beta-lactam drugs resistance. AmpC beta-lactamases produce resistance to various clinically important cephalosporins. The purpose of this study was to determine the frequency of AmpC beta-lactamase producing Escherichia coli and its occurrence in different clinical samples. This cross sectional observational study was conducted during April 2011 to March 2012 in the Microbiology Department of The Children's Hospital and Institute of Child Health, Lahore. A total number of 20,257 pathological samples like blood, cerebrospinal fluid [CSF], urine, pus, endotracheal tube [ETT], pleural fluid, sputum and urinary catheters were analyzed during the study period. E. coli were identified using API 20E system and resistance to cefoxitin was used as a screening test followed by disk potentiation as confirmatory test for AmpC beta-lactamases. E. coli were isolated in 670 samples out of which 85 [12.6%] were AmpC beta-lactamase producers. The gender distribution of patients with AmpC beta-lactamase producing E. coli was 52 [61.2%] in males and 33 [38.8%] in females. The occurrence rate of AmpC beta-lactamase producing E. coli in clinical samples was highest in blood 45 [52.9%].The present study shows moderately high frequency of AmpC beta-lactamase producing E. coli which may increase morbidity and mortality among children. Earlier detection of AmpC beta-lactamases will decrease the morbidity rate of AmpC beta-lactamase producing E. coli infection.

Frequency and antimicrobial susceptibility of acinetobacter species isolated from blood samples of paediatric patients

Acinetobacter species is a major nosocomial pathogen causing serious infections in immuno-compromised and hospitalized patients. The aim of this study was to determine the frequency and antimicrobial susceptibility pattern of Acinetobacter species in blood samples of paediatric patients. This cross sectional observational study was conducted during January to October, 2011 at The Children's Hospital and Institute of Child Health, Lahore. A total number of 12,032 blood samples were analysed during the study period. Acinetobacter species were identified using API 20E and their antimicrobial susceptibility pattern was studied using Kirby-Bauer disc diffusion method. The blood cultures showed growth in 1,141 cultures out of which 46 [4.0%] were Acinetobacter species. The gender distribution of Acinetobacter species was 29 [63.0%] in males and 17 [37.0%] in females. A good antimicrobial susceptibility pattern of Acinetobacter species was seen with sulbactam-cefoparazone [93.0%], imepenem and meropenem [82.6% each] and piperacillin-tazobactam [78.0%]. The antimicrobial susceptibility of cefixime [19.5%], co-amoxiclav and cefotaxime [23.9% each], cefuroxime [26.0%], ceftazidime and ceftriaxone [30.4%] was poor. The results of the present study shows high rate of resistance of Acinetobacter species with cephalosporins in nosocomial infections. The sulbactam-cefoperazone, carbapenems and piperacillin-tazobactam showed effective antimicrobial susceptibility against Acinetobacter species

Bacterial profile in catheter related infections among children

To establish the bacteriological profile of catheter related infections in patients having various indwelling medical devices. Descriptive study. The study was conducted in the Children Hospital and Institute of Child Health, Lahore during November 2005 to March 2006. A total of 100 culture positive samples from these patients were included comprising of 68 samples of Endotracheal Tubes, 14 samples of Peritoneal Dialysis Catheters, 11 samples of Urinary Catheters and 7 samples of Central Venous Catheters. The most frequent catheter colonizing bacteria were Pseudomonas spp. [34.7%], Klebsiella spp. [27.8%]. Escherichia coli [23.5%], Staphylococcus aureus [5.2%], Coagulase negative Staphylococci [3.5%], Proteus spp. [3.5%], Streptococcus pyogenes [0.9%] and non-haemolytic Streptococci [0.9%]. Catheter related infection remains an ongoing problem which cannot be completely eradicated; however by basic rules of hygiene it can be minimized

Bacteriology of infected wounds - a study conducted at children's hospital Lahore

The objective of the study was to determine the commonest bacteria associated with wound infections. Descriptive study. This study was conducted at The Children's Hospital and Institute of Child Health, Lahore over a six months period from August 2005 to January 2006. A total of 100 positive culture samples from patients with mean age of 6.2+ 0.25 were analyzed. Patient history and clinical findings were collected on a pre-coded form. Pus samples or wound swabs were collected from infected wounds and were analyzed through culturing and biochemical methods for aerobic bacteria. A total of 109 bacteria were isolated from 100 samples with almost same frequency of Gram positive cocci 54 [49.54%] and Gram negative bacilli 55 [50.45%]. Most frequently isolated organism was S. aureus 45 [41.28%] followed by Pseudomonas species 20 [18.35%]. Wound infection remains an ongoing problem which cannot be completely eradicated. However by taking prompt control measures against the most commonly isolated organism and proper care of wound may lead to the minimization of wound infection Skin, the largest organ in the human body, plays a crucial role in the sustenance of life through the regulation of water and electrolyte balance, thermoregulation, and by acting as a barrier to external noxious agents including microorganisms, however, when the epithelial integrity of skin is disrupted, a wound results[1] Wound infections are one of the most common hospital acquired infections and are an important cause of morbidity and account for 70-80% mortality[2, 3] Development of such infections represent delayed healing, cause anxiety and discomfort for patient, longer stay at hospital and add to cost of healthcare services significantly[4]. The importance of wound infections, in both economic and human terms, should not be under-estimated[5]. In a study, on average, patients with a wound infection stay about 6-10 days more than if the wounds heal without infections[6]. This additional stay almost doubles the hospital cost that is equivalent to between œ1, 168 and œ2, 398[7]. Wound infections can be caused by different groups of microorganisms like bacteria, fungi and protozoa[8]. However, different microorganisms can exist in polymicrobial communities especially in the margins of wounds and in chronic wounds[9]. The infecting microorganism may belong to aerobic as well as anaerobic group[10]. Most commonly isolated aerobic microorganism include Staphylo-coccus aureus [31%], Coagulase-negative staphy-lococci [CoNS] [5%], Enterococci [5%], Escheri-chia coli [9%], Pseudomonas aeruginosa [14%], Klebsiella pneumoniae [3%], Enterobacter species [9%], Proteus jnirabilis [3.5%], other streptococci [3%], Candida [1.3%] with 80% Candida albicans, Group D streptococci [2%] and Acinetobacter [2%]. Other gram-positive aerobes [2%] and anaerobes [2.7%] also cause wound infections[11]

Prevalence of bacteria in urenary tract infections among children

The aim of the study was to identify the most common bacteria responsible for the UTI's among the hospitalized children of various sex and age groups. The study was carried out in the Microbiology section of The Children's Hospital and Institute of Child Health, Lahore during July to October 2004. A total of hundred positive samples were included in this study. The pure growth of bacteria was isolated by inoculating the specimens on Cysteine Lactose Electrolyte Deficient [CLED] Agar. The organisms were identified with the help of biochemical testing. The most frequently isolated organisms in this study were E. coli and Klebsiella [both 37%], followed by pseudomonas [23], Proteus, Staphylococcus aureus and Acinetobacter 1% each]