Pseudomonas aeruginosa: evaluation of pathogen burden and drug-resistance trends in a tertiary care hospital

Objective: To evaluate the pathogen burden and antibiotic-resistance trends of Pseudomonas aeruginosa among hospitalised patients at a tertiary care hospital. Study Design: Retrospective, hospital record-based, cross-sectional study. Place and Duration of Study: Microbiology Laboratory, Allama Iqbal Medical College/Jinnah Hospital, Lahore, from January 2014 to December 2016

Methodology: A total of 5,960 samples were collected from clinically suspected cases of bacterial infections, admitted to the hospital. Microbial identification and antibiotic susceptibility pattern were carried out and analysed

Results: Out of a total of 5,960 samples, Pseudomonas aeruginosa was isolated from 1,268 [21.2%] specimens. Department-wise isolation rate was n=600 [42.9%], n=268 [15.4%], n=201 [12.6%], and n=199 [16.0%] from intensive care unit [ICU], surgical units, medical units, and Gynae wards, respectively [p<0.0001]. Sample-wise isolation rate was, wound swabs n=448 [35%], urine n=356 [28%], sputum n=187 [14 %], tracheal aspirate n=127 [10%], blood n=99 [7%], and broncho-alveolar lavage n=51 [4%] [p<0.0001]. Drug-resistance pattern showed low rates for carbapenems [meropenem n=440 [35%], Imipenem n=436 [34%] and beta-lactam + beta-lactamase inhibitor combination [piperacillin+ tazobactam n=437 [34%] while alarming rates were observed for cephalosporins [ceftazidime n=716 [56%], fluoroquinolones [ciprofloxacin n=690 [54%], cefoperazone+sulbactam n=685 [54%], aminoglycosides [gentamicin, n=669 [53%], amikacin n=608 [48%], and monobactams [aztreonam n=666 [52%]. Decreasing trend was observed only for amikacin 63% to 37%, aztreonam showed similar pattern throughout, while there was an increasing trend of drug resistance in all groups of antibiotics

Conclusion: Emerging drug-resistant strains of Pseudomonas aeruginosa are probably linked to the injudicious use of antibiotics, leading to ineffective empirical therapy. Therefore, we suggest that culture and antimicrobial susceptibility testing should be done for targeted antimicrobial therapy against Pseudomonas aeruginosa

False negativity of ziehl-neelsen smear microscopy: is the scale-up the worth it in developing countries?

Objective: To evaluate the false negative results of Ziehl-Neelsen [ZN] smear microscopy

Study Design: Descriptive study

Place and Duration of Study: Mycobacteriology Laboratory, Allama Iqbal Medical College [AIMC] and Jinnah Hospital, Lahore [JHL], Pakistan, from February 2014 to August 2016

Methodology: A total of 3,951 [pulmonary 2,773 and extra-pulmonary 1,178] samples were collected from strong TB suspected patients attending JHL Lahore. Follow-up cases were excluded. Every specimen was processed for ZN smear microscopy, Lowenstein Jensen [LJ] culture. SPSS 21.0 was used; false negative and positive results of ZN smear were calculated keeping LJ culture as gold standard

Results: Out of total 3,951 samples, sputum was most freqently found pulmonary sample 48.4% [n=1915], extrapulmonary samples, pleural fluid and pus samples were most commonly observed samples 12.0% [n=476] and 8.3% [n=329], respectively. Overall false negativity was 23.1% [pulmonary=19.6%, extra-pulmonary=29.2%] [p<0.001], Maximum false negative results were observed in pericardial, synovial, pleural fluids, and pus samples as 40.0%, 38.0%, 33.0% and 32.0%, respectively

Conclusion: ZN smear microscopy is not a very efficient tool in case of patients with the low mycobacterial load. Therefore, National TB Control programs should consider extending their diagnostic approaches from ZN microscopy to more advanced techniques

Acinetobacter Spp: Resistance and therapeutic decisions at the turn of the novel millennium

This study was planned to evaluate sample wise isolation and antimicrobial resistant trends of Acinetobacter spp in different departments of a tertiary care hospital. This was a transversal descriptive study, carried out in the clinical microbiology laboratory of the Allama Iqbal Medical College/ Jinnah Hospital, Lahore, Pakistan, during the period of January 2015 to December 2016. Every clinical specimen was processed for bacterial culture and antimicrobial susceptibly testing. A total of 3590 [2015=1780, 2016=1810] clinical specimens were processed. Of the total, only 54.7% were gram-negative, among these Acinetobacter spp were isolated from 10.1% and 16.5% samples respectively in 2015-16 with an overall rate of 24.3%. The highest occurrence of Acinetobacter spp isolates was reported from Intensive care units [ICU] [54%] followed by surgical units [25%] and medical units [16%]. It is noteworthy that ICU and internal medicine showed the highest resistance rates, whereas, lower resistance rate was observed for the outdoor patients [OPD]. Although collistin showed 0% resistant while ceftriaxone, ciprofloxacin, gentamicin, and tigecycline showed 90%, 68%, 66%, 66% and 62% resistance against Acinetobacter spp. respectively. An alarming increase in the resistance rate of meropenem, cefoperazone/sulbactam, piperacillin/ tazobactam, ciprofloxacin, and imipenem was observed from the year 2015 to 2016. This startling resistance acquired by Acinetobacter spp. within a period of one year, represent very limited therapeutic options left for the infections caused by Acinetobacter spp. Unavailability of effective drugs and limited therapeutic options enforce the health care practitioners to prescribe expensive and broad range antibiotics, which may cause harm to the patient. Therefore, it is need of an hour to better understand the antimicrobial patterns and optimize antimicrobial prescription policies for the control of multidrug-resistant Acinetobacter spp

Impact of putative bacteriocins against multidrug resistant clinical isolates

Objective: To use one of such mechanism like bacteriocins, produced by Lactobacilli activities against pathogens

Study Design: Analytical / observational study

Place and Duration of Study: This study was carried out at Gulab Devi Chest Hospital, Lahore from November 2012 to January 2013

Materials and Methods: This study included 203 clinical samples. Multidrug resistant clinical isolates were selected on the basis of their MAR [Multiple antibiotic resistances] index, antibiotic susceptibility testing, methicillin resistant Staphylococcus aureus [MRSA] with oxacillin disc, double disc synergism and combination disc test. Plasmid isolation, conjugation was performed. Well-Diffusion assay was used for screening of putative bacteriocins produced by Lactobacillus strains against MDRs. Physiological characterization of antimicrobial compounds and protein estimation was analyzed

Results: Twenty five strains were selected based on MAR index [>0.2]. In which 6 MRSA and 19 extended spectrum beta-lacatmases [ESBL] producers were further proceeded for antimicrobial activity with putative bactericins. Plasmid was easily transferred their resistance by the process of conjugation. Five bacteriocins were obtained from Lactobacillus strains isolated from commercial products. These bacteriocins showed a strong antibacterial activity against selected MDRs. Decrease in zone sizes was observed when putative bacteriocins were treated with heat, SDS [Sodium dodecyl sulfate] and Protinase k. Putative bacteriocins produced by Lactobacilli exhibit significant antibacterial activity against MDRs. SA1 has high antibacterial activity with high protein content of 13mg/ml

Conclusion: Putative bacteriocins produced by Lactobacilli exhibit significant antibacterial activity against selected MDRs

MDRs have ability to transfer their resistance to other bacteria. The peptidal component of these bacteriocins can be used as an alternative therapy. Proper hospital policies require minimizing the horizontal spread of MDRs. Hence, it is necessary to purify the antibacterial molecule out of putative bacteriocin for further analysis

Comparison of conventional TB diagnostic techniques with PCR IS6110 in tertiary care chest hospital Lahore Pakistan

Objective: To compare the efficiency of conventional diagnostic techniques and Insertion Sequence [IS]6110 based PCR assay for M. tuberculosis in pulmonary and extra-pulmonary specimens from tertiary care chest hospital

Study Design: Observational study

Place and Duration of Study: This study was conducted at Gulab Devi Chest Hospital, Lahore from August to January 2013

Materials and Methods: A total of 1599 [1417 pulmonary and 182 extra-pulmonary] non-duplicate clinical specimens, obtained over a period of six months, were tested by conventional techniques such as Ziehl-Neelsen staining [ZN], Lowenstein Jensen [LJ] medium and Fluorescent staining. MTB was extracted through DNAzol method. Insertion Sequence [IS] 6110 based PCR assay was used for M. tuberculosis from pulmonary and extra-pulmonary specimens. Of the 1599 specimens, 781 were suspect cases while 818 were MDR [follow up] cases. Mean age of TB patient was +/- 33 years. 18% of follow-ups and 20% of suspects were <20 year in age, 52% follow-ups and 36% suspects were about 20-40 years, and 30% follow-ups and 33% suspects were >40 years ofage

Results: It was seen that, among MDR cases [follow-ups] 68% were males and 32% were females. Similarly,among TB-suspects, 58% were males and 42% were females. Of total 168 suspected pulmonary samples ZN [48.2.7%], fluorescent microscopy [79.7%], LJ culture [52.9%] and PCR [91.6%] were positive for M. tuberculosis. In total 143 suspected extra-pulmonary samples, ZN [34.95%], fluorescent microscopy [45.5%], LJ culture [39.8%] and PCR [87.4%] werepositive

Conclusion: In contrast to conventional methods of TB diagnosis, PCR is more quick, sensitive, reliable and cost effective technique

Early diagnosis of hypertension with molecular genetic techniques

To standardize the molecular techniques for early diagnosis of genes of hypertension. These techniques include extraction of DNA in which we extracted DNA by manual method with results of high yield, less purity and it was more time consuming. On the other hand with kit [Fermentas] method yield was low with high purity and less time consuming. The purity of DNA was checked by spectrophotometer by using DNA/RNA ratio. Conditions for diagnosis was optimized for specific DNA sequence by using primers for genes Agt and Ace. Restriction digestion was done with the amplified product with restriction enzymes Lwel and Nco1 the result was found negative with no polymorphism. Single stranded conformational polymorphism [SSCP] was performed which is more efficient method of obtaining information about level of polymorphism with in anonymous nuclear loci than the restriction enzyme protocol. SSCP is also more specific because it gives idea which specific portion of gene is highly polymorphic. The result was again negative. This preliminary study of molecular analysis optimized the conditions for detection of polymorphism of candidate genes associated with hypertension. Following the same standardized conditions this is possible that we can study and diagnose thousands of hypertensive patients. This is very much helpful for future planning of persons who are prone to hypertension due to family history