Species identification and antibiotic susceptibilities of coagulase o negative staphylococci isolated from urinary tract infection specimens

Objective: To determine the incidence of Coagulase - negative S. aureus in urinary tract infections and sensitivities of these isolates to antimicrobial agents

Study Design: Cohort study

Place and Duration of Study: Dr. Essa Laboratory and Immunology and Infectious Disease Research Laboratory [IIDRL], Microbiology Department, University of Karachi, from January 2009 to January 2010

Methodology: Urine specimens, suggestive of urinary tract infection [UTI], were identified. Speciation of isolates was done using API-20 Staph. system. Screening of extracellular products was done using SDS-PAGE electrophoresis and Hemolysin on blood-agar plates. Minimum inhibitory concentration [MICs] of antibiotics was estimated by microtiter well plate method. Frequency and percentages were determined and chi-square test was used for comparing proportions with significance at p < 0.05

Results: Coagulase - negative S. aureus [CONS] were the cause of urinary tract infection in 56 out of 1866 outpatient [3%] and 164 of 1261 inpatient [13%], urinary tract infections [p < 0.001]. Two hundred and twenty CONS isolates were identified. The most common CONS identified was S. saprophyticus [31%, 68 strains]. The relative frequency of Coagulase - negative S. aureus was 6% [13 strains]. All isolates were sensitive to Vancomycin and Linezolid. Resistance was 69% to Ampicillin, 53% to Methicillin, and 37.5% to Ciprofloxacin

Conclusion: CONS are a potential uropathogens, with capability of slime production and resistance to common empirical prescriptions. This also warrants formulation of an appropriate antibiotic policy that covers CONS

Antibiotic resistance pattern of Pseudomonas aeruginosa isolated from urine samples of urinary tract infections patients in Karachi, Pakistan

The aim of this study was to evaluate the antibiotic resistance pattern of Psedomonas aeruginosa and its prevalence in patients with urinary tract infections [UTI] for effective treatment in a developing country like Pakistan. This is an observational study conducted for a period of ten months which ended on December 2013 at the Dr. Essa Laboratory and Diagnostic Centre in Karachi. A total of 4668 urine samples of UTI patients were collected and standard microbiological techniques were performed to identify the organisms in urine cultures. Antibiotic susceptibility testing was performed by Kirby-Bauer technique for twenty five commonly used antimicrobials and then analyzed on SPSS version 17. P. aeruginosa was isolated in 254 cultures [5.4%]. The most resistant drugs included Ceclor [100%] and Cefizox [100%] followed by Amoxil/Ampicillin [99.6%], Ceflixime [99.6%], Doxycycline [99.6%],Cefuroxime [99.2%], Cephradine [99.2%], Cotrimoxazole [99.2%], Nalidixic acid [98.8%], Pipemidic acid [98.6%] and Augmentin [97.6%]. Emerging resistant strains of Pseudomonas aeruginosa are potentially linked to injudicious use of drugs leading to ineffective empirical therapy and in turn, appearance of even more resistant strains of the bacterium. Therefore, we recommend culture and sensitivity testing to determine the presence of P.aeruginosa prior to specific antimicrobial therapy

Seasonal variation and identification of plasmodium species in blood samples of patients referred to labs of Karachi for suspected malaria from 2009 to 2011

To identify the seasonal effect on the frequency of malaria and its species in blood samples of patients referred to private and public Labs of Karachi for suspected Malaria and to observe if there has been any change in the frequency of the malaria species from May 2009 to Feb 2011. A cross sectional retrospective review of the laboratory data of the positive blood samples of malarial parasites over a period of 22 months from May 2009 till Feb 2011 via convenient sampling from a private and a public sector laboratory. Blood samples of patients of all ages and gender who were referred from various doctors with suspected diagnosis of malaria were checked for the presence of malarial parasite [MP] using the slide method and/ or Immunochromatographic Technique. The samples found positive for the MP were then analyzed for species identification. The seasonal variation in the frequencies of these species assessed and the number of positive MP results observed from May 2009 to Feb 2011 were described. Plasmodium vivax was found to be the most frequently identified plasmodium species present in 78.6% of the cases followed by Plasmodium falciparum. Adult males predominated over female and children. Frequency of Malaria was highest in the months of September with an overall surge of cases in the months of Aug to Dec. It is also observed that the number of cases in the year 2010 is much higher than in 2009. The study concludes that P.vivax is the most frequently isolated species causing malaria in Karachi. The frequency of P.vivax causing malaria seems to have increased from 2009 to 2010. Malaria was more frequent in the months of August to December in Karachi affecting adult males more frequently as compared to females and children

Current efficacy of antibiotics against Klebsiella isolates from urine samples - A multi-centric experience in Karachi

Due to emergence of bacterial resistant strains, the effectiveness of current antibiotic treatment without culture/sensitivity testing is questionable. Our study aims to assess the present sensitivity profiles of Klebsiella isolates from urine samples and provide options for empiric prescription in critically ill patients. Klebsiella pneumoniae isolates collected over a period of 28 months till January 2011 from 1, 617 urine samples of subjects presenting with Urinary Tract Infections were identified at a local diagnostic lab using standard protocol and subjected to Kirby-Bauer disk diffusion sensitivity testing. MICs were also estimated by E-nephelometry. Among 20 drugs used, low sensitivity was found to amoxicillin [0.1%], doxycycline [11.5%], nitrofurantoin [15.5%], amoxiclav [18.2%], gentamicin [35.4%], pipemidic acid, cephradine [40.3%] and cotrimoxazole [43.1%]. The isolates were more sensitive to cefuroxime [55.9%], cefixime [57.7%], ciprofloxacin [62.5%], ofloxacin [63%], ceftriaxone [66.2%], ceftazidime [66.4%], cefotaxime [66.6%], fosfomycin [77.5%] and amikacin [89.4]. Most effective were cefroperazone.sulbactam [95.8%], piperacillin.tazobactam [95.7%] and imipenem [97.7%]. Self-medication, lack of awareness, and the misuse of antibiotics by doctors has exacerbated the menace of microbial resistance. The study warrants the prudent choice of drugs in adherence with prevailing sensitivity profiles

Current pattern of antibiotic resistance of clinical isolates among conjunctival swabs

To identify the etiological agent in bacterial conjunctivitis and to determine the antibiogram of bacterial isolates. This observational study was conducted at Dr. Essa's Laboratory over a period of 12 months ending in March 2012. Two hundred samples taken from conjunctiva of patients with conjunctivitis were cultured on routine medium and the antibiograms of bacterial isolates were determined by Kirby- Bauer disc diffusion method. The analysis of the culture showed that 41% were cultured positive with gram positive bacteria Staphylococcus aureus 52.5% and Staphylococcus epidermidis 30.1% and Micrococci 8.3%. However, 9.1% were gram negatives with Klebsiella pneumoniae 5.14% and Pseudomonas aeruginosa 2.6% and 1.36% were others [Acinetobacter, Haemophilus, E.coli and Moraxella] keeping in view the increasing use of contact lens and unclean fingers. The overall antibiograms of bacterial isolates indicate aminoglycosides [gentamicin, tobramicin] and the newer quinolones as apparent drug of choice for empirical therapy, followed by chloramphenicol, since drug fussy gram-negatives such as Pseudomonas, Acinetobacter and E.coli were among the conjunctival isolates. Resistance profile of gram positive isolates shows cefixime 91.4%, doxycycline 57.9%, cotrimoxazole 29.3%, ampicillin 22.9%, ciprofloxacin 13.4%, cephradine 8.3%, cefuroxime 7.1%, fosfomycin 4.7%, ceftriaxone 3.6%, co-amoxiclav 3.6%, cefotaxime 3.5%, vancomycin 2.6%. Resistance to all conventionally used antibiotics is increasing, therefore identification of etiological agent and antibiogram is important to treat conjunctivitis and to avoid complications

Status of first-line anti-TB drugs: an audit of 84 clinical sputum AFB isolates in Karachi

To evaluate the current incidence of multidrug-resistant tuberculosis [MDR-TB] among positive sputum culture isolates in two commercial laboratories in Karachi, Pakistan. In this laboratory-based study eighty four AFB smear-positive sputum culture isolates grown on routine Lowenstein-Jensen [LJ] medium in two separate diagnostic labs during 12 months ending August 2011 were identified and subjected to antimicrobial susceptibility testing using LJ medium and anti-TB First-Line drugs [FLD]. MICs of the control H37RV-NCTC strain 7416 were compared with the test strains inoculated with the batch tested. Results were evaluated by the traditional resistance ratio method. The sputa expectorated by 45 females [53.57%] and 39 males [46.43%] aged 15-58 years yielded 84 M. tuberculosis isolates. The percentages of FLD resistance were Rifampicin [48.8%], Streptomycin [28.57%], Ethambutol [7.14%], and Isoniazid [4.76%]. Kanamycin [28.57%] was also tested. Only 12 [14.28%] of these were sensitive to all the FLDs, and 4 [4.76%] were MDR-TB strains [indifferent to Rifampicin and Isoniazid]. The sparse labs in Karachi that do AFB-cultures should periodically assess and publicize the frequency of MDR-TB isolates to guide empirical therapy. Rifampicin, a consistent part of the initial drug regimen for empirical prescription, was currently least effective on the strains encountered

Enteric fever in Karachi: current antibiotic susceptibility of salmonellae isolates

To determine the current sensitivity and resistance profile of Salmonellae [S.] isolates in a laboratory setting. An observational study. Dr. Essa's Laboratory and Diagnostic Centre, Karachi, Pakistan, from November 2008 - October 2010. Isolates from blood culture specimens of 481 bacteraemic patients were identified using conventional biochemical tests. Salmonellae was confirmed with specific antisera and their antibiograms determined by Kirby-Bauer Disc Diffusion method using 12 relevant antibiotics. Inclusions of the study were bacteraemia documented in all blood samples positive for S. typhi, S. paratyphi-A and B. Exclusions were all samples other than blood and blood samples negative for S. typhi and S. paratyphi-A and B during the same period. Multidrug resistance [MDR] of isolates was defined as the isolates showing resistance to all conventional anti-typhoid medicines i.e., Chloramphenicol, Ampicillin and Co-trimoxazole. Specimens [n=217] yielded 131 Salmonellae typhi [60.36%], 71 S. paratyphi-A [32.71%], and 15 S. paratyphi-B [6.9%]; these were sensitive to the Quinolones [Enoxacin: 94.96% [n=91], Ciprofloxacin, 96.47% [n=182], Ofloxacin: 95.74% [n=203]], and Cephalosporins [Cefixime: 96.62% [n=202], Cefotaxime: 99.17% [n=206], Ceftriaxone: 98.79% [n=208]]. Resistance to Amoxicillin was 96.48% [n=128] and 29.91% [n=78] to Co-trimoxazole. About 62.64% [n=136] of the isolates were MDR strains. Ciprofloxacin is currently a suitable empirical choice in presumed enteric fever cases, but culture and sensitivity analysis should be encouraged and results incorporated in prescription strategy. Increasing frequency of S. paratyphi-A isolates possibly suggests incomplete coverage employing monovalent vaccine

Association of BMI with testosterone in infertile males among local population

To ascertain association of raised BMI with Testosterone levels among infertile males. A cross sectional study was conducted from April 2010 to March 2011. Sample size was 300 males [147 infertile and 153 healthy fertile control subjects] aged 30 and 60 years, selected from the Jinnah Postgraduate Medical Center [JPMC], Aziz Medical Center and Abbasi Shaheed Hospital, Karachi. All the subjects were grouped to BMI criteria [Ascian pacific region for WHO]. They were selected by purposive sampling after a detailed medical history and physical examination. Semen analysis was done and blood samples were collected for serum Testosterone levels. Data was analyzed via SPSS 16.0 by using Analysis of Variances [ANOVA] and independent t test to compare the means and to observe sensitivity of tests and to evaluate the significant association with in the group. A significant negative correlation existed between Testosterone and BMI .Raised BMI appear to have an association with male infertility with the reduction in Testosterone levels. Obesity is confirmed as risk factor for male infertility in local population

Frequency of HBsAg positivity: a laboratory data analysis

The objective of the study was to evaluate the current frequency ratio of HBsAg seropositivity in a laboratory-based setting. It was a retrospective study, conducted during September 2009 and November 2010 at Dr. Essa's Diagnostic Laboratory and its branches in Karachi, Pakistan. Blood samples of 8,648 individuals were screened for HBsAg using the BioRad EVOLIS EIA method. Pertinent questionnaires filled in by 50 practicing physicians were also evaluated. The 551 subjects [6.4%] that tested positive for HBsAg included 360 males [65.3%] and 191 females [34.6%]. The highest frequency of HBsAg was noted in those aged 16-29 years [43.9%]. HBsAg rate in the studied population has steadily increased and is more common in the sexually active age group, with males tending to be more often seropositive than females

Ear infections in Karachi: the frequency and antibiotic resistance of bacterial isolates

This 12-month retrospective study was undertaken to determine the frequency of ear infections in children and adults, and the sensitivity of bacterial isolates to antibiotics available for their treatment. Ear swabs of 197 subjects with ear infections were processed in a local lab with branches in key areas of Karachi City from January 2009 to December 2009. The isolates from ear discharge were identified on conventional basis and their sensitivity to 24 antibiotics was determined by the Kirby-Bauer Agar Disc Diffusion Method. Among the patients, 100 were females and 97 males; infections were more common in children between 1 to 10 years. Staphylococcus aureus and Pseudomonas aeruginosa were mostly isolated among 10 bacterial species; Klebsiella pneumoniae was more often grown from female infected ears. The most effective antibiotics determined for possible empirical prescription included Pipericillin+Tazobactam, Cefoperazone+Sulbactam, Imipenam, and Fosfomycin. While Ciprofloxacin [57.7%] and Amoxyclav [36.2%] exerted transitional activity, a majority of the isolates were indifferent to Cotrimoxazole, Cefixime, Lincomycin, Doxycycline and Polymyxin B. The increasing resistance of causative organisms in our environment to multiple antibiotics encourages the C/S of specimens prior to drug prescription in order to reduce the chances of treatment failure and amplified antibiotic resistance. When unavoidable, however, a selection of drugs is offered for empirical preference

Current pattern of antibiotic resistance in stapbylococcus aureus clinical isolates and the emergence of vancomycin resistance

To determine the pattern of antibiotic resistance in the clinical isolates of Staphylococcus [S.] aureus, Methicillin Resistant S. aureus [MRSA] and define the possible emergence of Vancomycin resistant S. aureus [VRSA] in Karachi. An observational study. Essa Laboratories and Department of Molecular Genetics, Ziauddin Hospital, from January to December 2009. Staphylococcal isolates from different clinical specimens, pus, urine, blood, high vaginal swab and other secretions received at Ziauddun laboratories and Dr.Essa laboratories were collected. The specimens were inoculated on blood agar, MacConkey agar and Chrom agar. Antibiotic susceptibility to conventional antibiotics was done by disc diffusion, and E-test. Methicillin resistance was tested by using Oxacillin and Methicillin disks and confirmed by gold standard PCR for presence of mecA gene. All MRSA strains were subjected in addition to Vancomycin screen agar test. Out of the 450 S. aureus isolates 174 [38.6%] were found to be MRSA. In those isolates, high resistance was found to Cefixime [100%] Doxicycline [100%] Oxacillin [96.5%] Gentamicin, [96.3%], Timethoprim/Sulfametoxazole [95.6%] Chloramphenicol [93%] Tobramicin [81.03%], Ofloxacin [72.4%] and Ciprofloxacin [63.7%]. Low resistance was found to Ceftazidine [36%], Amoxicillin/Clavulanate [32.7%], Fosfomycin [31%], Cefroxime [24%], Amikacin [17.2%] and Meropenem [13%]. One isolate was found to be Vancomycin resistant [MIC 32 microg/ml]. Four isolates had intermediate resistance, with two strains having MIC of 16 microgram/ml and two having MIC of 8 microgramg/ml. These strains were also resistant to all the other tested antibiotics except Linezolid to which all isolates were susceptible. Antibiotic resistance to all the conventionally used antibiotics was high in the tested isolates. All the strains were susceptible to Linezolid which is an expensive alternative with adverse side effects. Judicious use of antibiotics focused on the compliance and formation of antibiotic policy guide lines is highly recommended

Current pattern of bloodstream infections in a tertiary care hospital of Karachi and clinical significance of positive blood cultures

Blood stream infections [BSI] [septicemias] require prompt empirical therapy based on awareness of the drug susceptibility profiles of locally prevalent pathogens isolated. Department of Pathology Dow University of Health Sciences, Karachi, in collaboration with Dr Essa's Diagnostic Centre, from July to November 2008. A cross-sectional prospective study. 324 consecutive blood cultures from patients coming to Civil Hospital Karachi were scrutinized for bacterial isolates and their antibiotic sensitivity profiles were done. A total of 100 [30.9%] specimens were found positive, of these 78% isolates were gram negative bacteria and 22% gram positive cocci. Salmonella typhi, seen less often in adults [20.5%], was the predominant pathogen in children [82.5%]. Escherichia coli, Staphylococcus aureus, Pseudomonas and Klebsiella were isolated in neonates, and Klebsiella, Pseudomonas, Enterobacter and Acinetobacter were identified among adult patients. While all S.typhi isolates were sensitive to cefixime, ceftriaxone and the fluoroquinolones and increasingly amenable to chloramphenicol, of significance was the percentage of other multidrug resistant bacterial isolates. Methicillin resistant Staphylococcu aureus was isolated from one case. The results were analysed by applying SPSS version 16 to derive p value. Amikacin, carbapenems, cefoperazone+sulbactam, fosfomycin and pipericillin+tazobactam are currently the only available drugs still active in-vitro on blood isolates, judicious use of antibiotics focused on the compliance and formation of antibiotic policy guide lines is highly recommended

Complete androgen insensitivity syndrome

The incidence of Complete Androgen Insensitivity Syndrome [CAIS] is about 1 in 20.000. People with CAIS are normal appearing females, despite the presence of testes and a 46, XY chromosome constitution. We came across a case in which a 17 years old girl presented with the complaint of inguinal hernia and amenorrhea. Subsequent investigations were done revealing absence of female internal genitalia and the presence of abdominal mass, possibly testes. Syndrome has been linked to mutations in AR, the gene for the human Androgen Receptor, located at Xq11-12 leading to the insensitivity of the receptor to testosterone. Gonadectomy was performed and life long Hormone replacement therapy was advised