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The study was undertaken to determine the in-vitro antibacterial potential of Diospyrosblancoi, Phoenix dactylifera and Morusnigra leaf extracts in hexane, chloroform, methanol, ethyl-acetate and aqueous extracts against dental caries causing bacteria. Disc diffusion assay was used to determine the antibacterial efficacy; the extracts were further separated using Thin Layer Chromatography and the anti-biofilm activity of the extracts was also determined. The preliminary phytochemical screening of the extracts revealed the presences of flavonoids, saponins, alkaloids and tannins because of which the extracts showed strong antibacterial activity against the selected pathogens. The ethylacetate extracts showed maximum inhibitory effect on biofilm formation by S. mutans.96% inhibition was observed in methanol extract of Diospyrosblancoi, and 95% in ethyl acetate extract. The results evidenced that the plants inhibit the growth of oral bacteria responsible for dental caries with their abundance source of secondary metabolites and can be used as an alternative treatment for caries, thus minimizing the antibiotics used to treat the disease in local medicine
Subject(s)
Anti-Bacterial Agents/pharmacology , Diospyros/chemistry , Phytotherapy , Plant Extracts , Plant Leaves , Plants, MedicinalABSTRACT
Methicillin resistant Staphylococcus aureus [MRSA] is resistant to known antibiotics and has become a great challenge for healthcare professionals, therefore new molecules are needed to manage this situation. In this study, new lead molecules 4-Amino-5-[2-Hydroxyphenyl]-l,2,4-Triazol-3-Thione [Ul] and4-[2-hydroxybenzalidine] amine-5-[2-hydroxy] phenyl-l,2,4-triazole-3-thiol[U!A Schiff base] were synthesized by fusion method that showed promising antibacterial activity [U1A: 26mm and Ul: 14mm] against MRSA.FT-IR and NMR were used for structural characterization of these derivatives and their toxicity properties were assessed by Lipinski's rule of 5. New potential drug targets of this bacterium were also identified by comparative and subtraction genomics techniques. In particular, octanoyl-[GcvH]: protein N-octanoyl transferase and phosphor mevalonate kinase were used as potential targets in AutoDock Vina studies. This study can provide a framework to find potential drug targets for other pathogenic microorganisms that can successfully be docked with compound Ul and Ul A
Subject(s)
Methicillin , Methicillin-Resistant Staphylococcus aureus , Drug Resistance, Bacterial , Delivery of Health Care , Drug DesignABSTRACT
Objective: To scrutinize patterns of multi-drug-resistant uropathogenic Escherichia coli (UPEC) strains and particularly of fluoroquinolone-resistance this is an alternative choice for the treatment of urinary tract infections. Methods: Bacterial samples (n = 250) were collected from out-patients from August 2012 to August 2014 Islamabad. Antibiotic susceptibility profiling and determination of minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations were performed according to the guidelines of Clinical and Laboratory Standards Institute (CLSI, 2012). Genes, qnrA, qnrB and qnrS were identified by DNA amplification and sequencing. Results: The highest percentage of UPEC isolates were resistant to co-trimoxazole (82%) followed by cephalothin (80%), 2nd Gen, 3rd Gen and 4th Gen cephalosporins, respectively. Resistance against gentamicin, amikacin remained 29% and 4%. For other drugs including nitrofurantoin, tetracycline, carbapenem and beta-lactam inhibitors remained below 10%. Altogether, 59% of the isolates were resistant to at least three antibiotics including one fluoroquinolone. Overall, MICs for ciprofloxacin remained (MIC ≥ 256 μg/mL) and for levofloxacin (MIC ≥ 16 μg/mL and 32 μg/mL). No significant differences were observed regarding MIC values of extended spectrum β-lactamase (ESBL) and non-ESBL producers. For qnrS and qnrB positive isolates MICs remained above 32 μg/mL. Prevalence of UPEC was significantly higher among females and 40% of the isolates were ESBL producers. Conclusions: Higher percentages of ESBL producing UPEC were associated with urinary tract infections. Moreover, the majority of these isolates were multi-drug resistant and fluoroquinolone-resistant.
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OBJECTIVE@#To evaluate current situation regarding the prevalence of hepatitis C virus (HCV) in thalassemic patients visiting a thalassemia centre in Rawalpindi District, Pakistan for supportive therapy.@*METHODS@#Serum samples were screened for hepatitis B surface antigen and anti-HCV by using commercially available ELISA kit. Micro-plate reader was used to perform analysis based on the absorbance/cut-off ratios. Samples were considered positive or negative. Results from ELISA were analyzed statistically.@*RESULTS@#A total of 95 subjects were observed to have β-thalassemia major (96%) and β-thalassemia intermedia (4%). Among these, 47 (49%) were detected positive for anti-HCV antibodies and three for hepatitis B surface antigen. All recruited subjects were observed for therapy/medication behavior and clinical complications. About 83 (87%) patients were on chelation therapy, and overall complications (hepatomegaly, splenomegaly and splenectomy) were observed in 81% individuals. The distribution of disease status (thalassemia and hepatitis) was not significantly associated with gender, age, origin, province, socio-economic status and parental marriage type (P>0.05). The distributions of ferritin levels, therapy/medication and complications were assessed across demographic variables. Thalassemic subjects were distributed with respect to their sporadic and familial presentations. Among the familial cases (n=35), a total of 93 subjects were found to be affected. Parity was scored for the index cases, and majority belonged to second parity (29%), followed by first and third parities (25% and 15%, respectively). The sibship size was increasing with increasing parity level.@*CONCLUSIONS@#Although standardized blood screening procedures are supposed to be implemented, higher prevalence of HCV in thalassemic patients requires greater attention in Pakistan. Furthermore, a poor compliance regarding iron chelation therapy has been observed in this study.
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<p><b>OBJECTIVE</b>To evaluate physico-chemical properties and antimicrobial potential of indigenous honey samples against different reference strains including Escherichia coli ATCC 8739, Enterobacter aerogenes ATCC 13048, Pseudomonas aeroginosa ATCC 9027, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Salmonella typhi ATCC 14028, Klebsiella pneumonia ATCC 13883, Aspergillus niger ATCC 16404, Rhizopus oligosporus PCSIR1, Candida albicans ATCC 14053 and Candida utilis ATCC 9950.</p><p><b>METHODS</b>By using standard methods samples were evaluated for their antimicrobial properties including additive effect of starch and non-peroxidase activity, antioxidative properties (phenol contents, flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity). Prior to this evaluation, complete physico-chemical properties including pH, color, ash contents, protein contents, moisture contents, hydroxymethyl furfural contents, total sugar contents, reducing sugar and non-reducing sugar contents were analyzed.</p><p><b>RESULTS</b>Relatively higher ash contents were found in the Siddar honey i.e. (0.590 0±0.033 6)% and small honey showed relatively higher protein contents i.e. (777.598±9.880) mg/kg. The moisture contents of tested honey samples ranged between 13.8%-16.6%, total sugar contents from 61.672%-72.420% and non-reducing sugar contents from 1.95%-3.93%. Presences of phenolic contents indicate higher antioxidant potential of these honey samples. All bacteria showed clear inhibition zones in response to tested honey samples whereas fungi and yeast showed inhibition at higher concentrations of these honey samples. For Escherichia coli, Bacillus subtilis, Salmonella typhi, Pseudomonas aeroginosa and Aspergillus niger, overall the small honey showed the higher activity than other honey samples.</p><p><b>CONCLUSION</b>Physico-chemical analysis of honey samples confirmed good quality of honey according to the standards set by European Union Commission and Codex Alimentarius Commission. Evaluation of these honey samples confirms antimicrobial potential of particular types of honeys indigenous to Pakistan.</p>
ABSTRACT
Objective: To evaluate physico-chemical properties and antimicrobial potential of indigenous honey samples against different reference strains including Escherichia coli ATCC 8739, Enterobacter aerogenes ATCC 13048, Pseudomonas aeroginosa ATCC 9027, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Salmonella typhi ATCC 14028, Klebsiella pneumonia ATCC 13883, Aspergillus niger ATCC 16404, Rhizopus oligosporus PCSIR1, Candida albicans ATCC 14053 and Candida utilis ATCC 9950. Methods: By using standard methods samples were evaluated for their antimicrobial properties including additive effect of starch and non-peroxidase activity, antioxidative properties (phenol contents, flavonoid contents, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity). Prior to this evaluation, complete physico-chemical properties including pH, color, ash contents, protein contents, moisture contents, hydroxymethyl furfural contents, total sugar contents, reducing sugar and non-reducing sugar contents were analyzed. Results: Relatively higher ash contents were found in the Siddar honey i.e. (0.590 0±0.033 6)% and small honey showed relatively higher protein contents i.e. (777.598±9.880) mg/kg. The moisture contents of tested honey samples ranged between 13.8%-16.6%, total sugar contents from 61.672%-72.420% and non-reducing sugar contents from 1.95%-3.93%. Presences of phenolic contents indicate higher antioxidant potential of these honey samples. All bacteria showed clear inhibition zones in response to tested honey samples whereas fungi and yeast showed inhibition at higher concentrations of these honey samples. For Escherichia coli, Bacillus subtilis, Salmonella typhi, Pseudomonas aeroginosa and Aspergillus niger, overall the small honey showed the higher activity than other honey samples. Conclusion: Physico-chemical analysis of honey samples confirmed good quality of honey according to the standards set by European Union Commission and Codex Alimentarius Commission. Evaluation of these honey samples confirms antimicrobial potential of particular types of honeys indigenous to Pakistan.
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Objective: To evaluate current situation regarding the prevalence of hepatitis C virus (HCV) in thalassemic patients visiting a thalassemia centre in Rawalpindi District, Pakistan for supportive therapy. Methods: Serum samples were screened for hepatitis B surface antigen and anti-HCV by using commercially available ELISA kit. Micro-plate reader was used to perform analysis based on the absorbance/cut-off ratios. Samples were considered positive or negative. Results from ELISA were analyzed statistically. Results: A total of 95 subjects were observed to have β-thalassemia major (96%) and β-thalassemia intermedia (4%). Among these, 47 (49%) were detected positive for anti-HCV antibodies and three for hepatitis B surface antigen. All recruited subjects were observed for therapy/medication behavior and clinical complications. About 83 (87%) patients were on chelation therapy, and overall complications (hepatomegaly, splenomegaly and splenectomy) were observed in 81% individuals. The distribution of disease status (thalassemia and hepatitis) was not significantly associated with gender, age, origin, province, socio-economic status and parental marriage type (P>0.05). The distributions of ferritin levels, therapy/medication and complications were assessed across demographic variables. Thalassemic subjects were distributed with respect to their sporadic and familial presentations. Among the familial cases (n=35), a total of 93 subjects were found to be affected. Parity was scored for the index cases, and majority belonged to second parity (29%), followed by first and third parities (25% and 15%, respectively). The sibship size was increasing with increasing parity level. Conclusions: Although standardized blood screening procedures are supposed to be implemented, higher prevalence of HCV in thalassemic patients requires greater attention in Pakistan. Furthermore, a poor compliance regarding iron chelation therapy has been observed in this study.
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Polypeptide antimicrobials used against topical infections are reported to obtain from mesophilic bacterial species. A thermophilic Geobacillus pallidus SAT4 was isolated from hot climate of Sindh Dessert, Pakistan and found it active against Micrococcus luteus ATCC 10240, Staphylococcus aureus ATCC 6538, Bacillus subtilis NCTC 10400 and Pseudomonas aeruginosa ATCC 49189. The current experiment was designed to optimize the production of novel thermostable polypeptide by applying the Taguchi statistical approach at various conditions including the time of incubation, temperature, pH, aeration rate, nitrogen, and carbon concentrations. There were two most important factors that affect the production of antibiotic including time of incubation and nitrogen concentration and two interactions including the time of incubation/pH and time of incubation/nitrogen concentration. Activity was evaluated by well diffusion assay. The antimicrobial produced was stable and active even at 55degree C. Ammonium sulphate [AS] was used for antibiotic recovery and it was desalted by dialysis techniques. The resulted protein was evaluated through SDS-PAGE. It was concluded that novel thermostable protein produced by Geobacillus pallidus SAT4 is stable at higher temperature and its production level can be improved statistically at optimum values of pH, time of incubation and nitrogen concentration the most important factors for antibiotic production.
Subject(s)
Anti-Bacterial Agents/biosynthesis , Research Design , Peptides/metabolismABSTRACT
The vacuolating cytotoxin VacA and cytotoxin associated gene product CagA, encoded by vacA and cagA are major virulence determinants associated with pathogenesis of Helicobacter pylori. The presence and prevalence of two major H. pylori virulence associated genes among gastric biopsies of Pakistani children were investigated in the current study. Fifty one gastric biopsy specimens of children were analysed for 16S rRNA, vacA and cagA genes using PCR. The results showed that 21 (41.2%) biopsies were positive for H. pylori as determined by 16S rRNA PCR. In the 21 H. pylori positive gastric biopsies, 19 (90.5%) showed vacA s1a, 1 (4.75%) was vacA s1b and 1 (4.75%) was vacA s2 whereas, 5 (23.8%) were vacA m1 and 16 (76.2%) were vacA m2. None of the H. pylori positive biopsies carried vacA s1c subtype. The cagA gene was found in 13 (61.9%) of H. pylori infected biopsies and different vacA combinations were found with or without cagA gene. H. pylori was detected with high frequency of cagA while vacA s1a and vacA m2 regions with vacA s1a/m2 genotype were predominant in H. pylori infected gastric biopsies of children.
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The global problem of increasing trend in antimicrobial resistance is particularly pressing in the developing countries, where the Methicillin-Resistant Staphylococcus aureus [MRSA] is often the severe casual agent in hospital-acquired infections. This multi-centre surveillance prospective study was planned to define the magnitude of problem of MRSA among clinical isolates from four teaching hospitals of Lahore Pakistan; Mayo, Services, Jinnah and Shaikh Zayed Hospitals during April 2006-March 2008. Identification of organisms was done by the standard Microbiology methods. MRSA isolates identified on Kirby-Bauer disc diffusion were further evaluated by minimum inhibitory concentration on BD PhoenixTM system and detection of mecA gene by pulsed-field gel electrophoresis [PFGE] PCR. Of the total 1,102 S. aureus isolates, oxacillin resistance was found in 462 on disc diffusion and 420 on MIC while mecA gene was detected from 307 strains. The prevalence of MRSA among S. aureus isolates was 41.9%, 38.1% and 27.9% on disc diffusion, MIC, and mecA gene detection respectively. Hospital acquired-MRSA strains were multi drug resistant while community acquired-MRSA showed susceptibility to clindamycin [63%], ciprofloxacin [24.2%] and SMZ/TMP [3.9%]. In diagnosing MRSA, the sensitivity and specificity rates of disc diffusion test were 100% and 83.7% while MIC 96.2% and 93.3% respectively. There is an increasing trend in emergence MRSA and the conventional method of antimicrobial susceptibility testing showed false positive tests. This is the reason of misuse of vancomycin by physicians which may further increase MRSA in Pakistan. Therefore, molecular diagnostic facilities are recommended to avoid falsesusceptible results
Subject(s)
Drug Resistance, Microbial , Microbial Sensitivity Tests , Staphylococcus aureus , Cross Infection , Prospective StudiesABSTRACT
The evaluation of resistance to ciprofloxacin [CIP] in Escherichia coli [n=1112] isolated from clinical specimens including urine, pus, high vaginal swabs, etc. was carried out at Rawalpindi General Hospital, Rawalpindi during the years 2002- 2005. The isolates were identified by colonial morphology, gram staining, and biochemical tests like indole, citrate and TSI. The antibiotic susceptibility of these isolates was determined by Kirby-Bauer disk diffusion method according to the standard guidelines of Clinical Laboratory Standards Institute [CLSI]. During the year 2002 resistance to ciprofloxacin was 49.19% [n=187], in the year 2003 it was 47.5% [n=297], in year 2004 was 59.03% [n=332], in year 2005 was 46.28% [n=296]. When resistant isolates from different sources were compared E. coli uropathogens varied between 45.2 to 58.53% during the four year. While pus isolates in 2003 were highly resistant [69% resistance] and isolates from high vaginal swab showed peak resistance in 2004 [66%] Fig. 3 table 4. In the year 2002, four fluoroquinolones [FQs] i.e. ofloxacin [OFX], ciprofloxacin [CIP], enoxacin [ENX] and sparfloxacin [SPX] were included in the study while norfloxacin [NOR] was tested against urinary isolates only. During that year 203 isolates were identified as E. coli; 124 from urine, 45 from pus, 21 from high vaginal swabs, and 13 from other sources. Highest resistance was seen in isolates from urine ranging from 54-68% against these FQs
Subject(s)
Humans , Drug Resistance, Bacterial , Escherichia coli/drug effects , Microbial Sensitivity TestsABSTRACT
The purposes of this study were to give the characteristics of Streptococcus mutans isolated from saliva, and the salivary level of S. mutans and its relationships with dental caries. Dental caries is a common infectious disease worldwide. The aetiology of the disease is multifactorial, life habits and streptococcus mutans infection being the most imporatent factors. Paraffin stimulated saliva was taken from 40 subjects of three age groups in sterilized glass tubes. The saliva sample was serially diluted on GSTB agar plates. The plates were incubated anaerobically at 37 °C for 48 hours in candle extinction jars. S. mutans were identified on the basis of morphologic and biochemical characteristics. Viable cell count of S. mutans [CPU I ml] was also done on GSTB agar plates. S. mutans were found in all samples in significant percentage. The results indicated that subjects that had dental caries harboured high level of S. mutans 10[6]> CPU I ml. In age group III, the percentage of subjects with high level of S.mutans and dental caries was found to be higher as compared to age group I and age group II.
Subject(s)
Humans , Streptococcus mutans , Dental Caries/etiology , Culture Media , Dental Caries/microbiologyABSTRACT
The objective of the study was to find the antimicrobial effect of local brands of toothpastes containing different active ingredients, against aerobic and anaerobic oral flora and Streptococcus mutans; the causative agent of dental caries and dental plaque. The efficacy of eight locally formulated dentifrices was checked against aerobic and anaerobic oral bacteria of 30 subjects. Sterilized swabs were applied on the subject's teeth and inhibitory zones against these bacteria were measured by agar diffusion method. The magnitude and duration of Streptococcus mutans percentage reduction was evaluated against 1:100 and 1:1000 w/v dilutions of six dentifrices containing different active ingredients. The Streptococcus mutans were treated with selected dentifrices for 0,5,10,15,30,60 and 120 minutes. The CPU was done using spread plate method. Triclosan containing toothpastes showed maximum inhibitory zones against aerobic and anaerobic oral bacteria followed by fluoridated, whitening and herbal dentifrices. The maximum percentage reduction of S. mutans was shown by triclosan containing toothpastes followed by fluoridated and herbal dentifrices. All dentifrices showed antibacterial activity against aerobic and anaerobic oral bacteria, maximum being that of triclosan containing toothpastes. The longest and largest reduction of S. mutans was shown by triclosan containing dentifrices
Subject(s)
Humans , Anti-Bacterial Agents , /microbiology , Streptococcus mutans , Dental Caries , Dental Plaque , Dentifrices , Triclosan , Fluoridation , Microbial Sensitivity TestsABSTRACT
To determine the prevalence of extended-spectrum bi-lactamases [ESBLs] among the bacteria of family Enterobacteriaceae isolated from Nosocomial and outpatients, with double disc diffusion / synergy test. The bacterial strains were isolated from pus, sputum, blood, urine, pleural fluid, peritoneal fluid and cerebro-spinal fluid samples, obtained from both Nosocomial and outpatients. The samples were obtained from patients admitted in oncology, post-operative surgical, kidney transplant center / urology wards and intensive care unit of Pakistan Institute of Medical Sciences as well as outpatients of the hospital. Bacterial isolates, of family Enterobacteriaceae, were obtained from 200 Nosocomial and 200 outpatients [ambulatory]. The isolates were sub-cultured, identified, and the double disc diffusion / synergy test was performed for detection of ESBLs. Double disc diffusion / synergy test, for the detection of ESBLs production in Enterobacteriaceae. Prevalence of ESBLs in the Enterobacteriaceae was found to be 37.50% in Nosocomial and 06% in outpatient isolates. Highest prevalence was seen in Klebsiella pneumoniae [70%], followed by Enterobacter cloacae [33.33%] and Escherichia coli [28.57%] in case of Nosocomial isolates while in case of out-patient [ambulatory] isolates, the Enterobacter cloacae are the most prevalent ESBLs producers [8.33%]. Conclusions: Prevalence of ESBLs among the bacteria of family Enterobacteriaceae was higher in isolates obtained from Nosocomial patients as compared to out-patient [ambulatory] isolates. Such type of antimicrobial resistance appears to be particularly influenced by irrational use of antibiotics. To overcome this problem, the combined competencies of clinicians, microbiologists and the infection control team are needed