Isolation and drug susceptibility pattern of uropathogens in Saudi diabetic and non-diabetic patients with urinary tract infection.

Urinary tract infection (UTI), contribute substantially to healthcare burden. Diabetes predispose to UTI with high glycosuria being fertile medium for bacterial growth. With changing bacterial drug resistance patterns; the problem needs to be studied periodically to ensure a rational therapy, minimize adverse effects, and cost. Therefore, it is of interest to compare the profile and susceptibility pattern of uropathogens isolated from diabetic and non-diabetic patients with UTI. Mid-stream urine samples of 1100 patients (diabetic and non-diabetic), presenting with UTI symptoms were aseptically collected and inoculated into CLED medium. Colony counts of 105cfu/ml or 104cfu/ml and >5 pus cells per high power microscopic field were regarded as significant bacteriuria. Colonies from CLED were sub-cultured onto sheep blood agar and MacConkey agar. Bacterial identification was performed on the basis of colony morphology, gram staining, and series of biochemical tests though Analytical Profile Index (API) test strips. Drug susceptibility was done by standard Kirby-Bauer disk diffusion. Data was analyzed by SPSS ver. 25.Clinically significant bacteriuria was 32.8% and 19.2% in diabetics and non-diabetics respectively. The frequency of male and female patients was 153 and 208 in diabetic group; and 69 and 142 respectively in non-diabetic group. Diabetics were twice at risk of UTI; [Odds ratio; 2.04 (CI: 1.68-2.48, p<0.05)]. .Escherichia coli and klebsiella were most common gram-negative, while Staphylococcus aureus and Coagulase-negative staphylococci (CoNS) were most common gram-positive bacteria in both the groups. Most effective antibiotics against gram-negative bacteria were carbapenems, amikacin, colistin, and piperacillin/tazobactam; while ampicillin/amoxicillin, fluoroquinolones and cephalexin were least effective. For gram-positives, vancomycin, linezolid and tigecycline were most effective. No significant difference in bacterial profile and susceptibility pattern was found between diabetics and non-diabetics. However, diabetics were twice at risk of UTI compared to non-diabetics.

In vitro antibacterial activity of honey against multidrug-resistant Shigella sonnei.

BACKGROUND AND PURPOSE: The health benefits of honey as an oral therapeutic agent for the treatment of diarrhoea caused by Shigella sonnei depend on the ability of honey to withstand human gastrointestinal conditions. This study aimed to investigate whether honey could withstand and inhibit the growth of Shigella sonnei under such conditions. MATERIALS AND METHODS: We initially evaluated the survival of Shigella sonnei in human simulated gastric conditions (SGC) and simulated intestinal conditions (SIC). This was followed by determination of the susceptibility of Shigella sonnei to Manuka and Talah honey under gastrointestinal conditions. The colony forming units (CFU) of Shigella sonnei and minimum inhibitory concentrations (MICs) of honey were calculated. RESULTS: Shigella sonnei was unable to survive in the acidic environment of the stomach without food matrix and survived only when inoculated with a food source, resulting in 1.5 × 105 ± 0.2 CFU at 60 min and 1.7 × 105 ± 0.3 CFU after 120 min of incubation. In SIC, it survived both with and without food matrix at the same CFU (1.2 × 107 ±0.4) at 60 min and 1.7 × 107 ±0.2 CFU after 120 min of incubation. Growth of Shigella sonnei was not observed in SGC in the presence of either honey at different concentrations without a food source. In the presence of a food source, Manuka honey inhibited the growth of Shigella sonnei at 10% v/v and Talah honey at 20% v/v dilutions in SGC. In SIC, Manuka honey inhibited the growth of Shigella sonnei at 15% and 20% v/v dilutions, whereas Talah honey inhibited Shigella sonnei at 20% and 25% v/v dilutions without and with food sources, respectively. CONCLUSION: Shigella sonnei can survive in the acidic environment of the stomach if inoculated with a food source. Acidic pH and pepsin had no deleterious effects on the antibacterial capability of honey. However, bile reduced the antibacterial activity of honey in the intestinal environment.

In vitro evaluation of methicillin-resistant and methicillin-sensitive Staphylococcus aureus susceptibility to Saudi honeys.

BACKGROUND: Honey has been increasingly recognized as a potential therapeutic agent for treatment of wound infections. There is an urgent need for assessment and evaluation of the antibacterial properties against wound pathogens of honeys that have not yet been tested. METHODS: Ten Saudi honeys collected from different geographical locations were screened initially for their antibacterial potential against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) by the agar well diffusion method. Manuka honey (UMF-12) was used for comparison. Of the tested honeys, the honey that exhibited the greatest antibacterial activity in the agar well diffusion assay was further evaluated for its minimum inhibitory concentration (MIC) against ten MRSA clinical isolates and three American Type Culture Collection (ATCC) reference strains by the microbroth dilution method. RESULTS: Locally produced honeys exhibited variable antibacterial activity against the tested isolates in the agar well diffusion assay. They were unable to exhibit antibacterial activity against MSSA and MRSA at 25% dilutions (w/v) in catalase solution. However, Sumra and Talha honeys showed a zone of inhibition at 50% dilutions (w/v) in catalase solution. This finding means that both honeys possess weak non-peroxide-based antibacterial activity. Moreover, Sumra honey showed a larger inhibition zone at 50 and 25% dilutions (w/v) in distilled water than Manuka honey against both MSSA and MRSA. This result demonstrates that Sumra honey has more hydrogen peroxide-related antibacterial activity or total antibacterial activity than Manuka honey. In addition, MIC results obtained through a microbroth dilution assay showed that Sumra honey inhibited the growth of all MRSA clinical isolates (n = 10) and reference strains [MRSA (ATCC 43300) and MSSA (ATCC 29213)] at lower concentrations (12.0% v/v) than those required for Manuka honey-mediated inhibition (14.0% v/v). This result means that Sumra honey has more peroxide or synergistic antibacterial activity than Manuka honey. An equivalent MIC (15.0% v/v) was observed for E. coli (ATCC 25922) between Manuka honey and Sumra honey. CONCLUSIONS: Sumra honey may be used as an alternative therapeutic agent for infected wounds and burns, where additional hydrogen peroxide-related antibacterial activity is needed. In the future, the physiochemical characteristics of Sumra honey may be evaluated and standardized.

Antibacterial activity of Pakistani Beri honey compared with silver sulfadiazine on infected wounds: a clinical trial.

OBJECTIVE: To determine the antibacterial activity of Pakistani Beri honey in patients with infected wounds in comparison with silver sulfadiazine. METHOD: Inpatients with infected wounds at a tertiary care hospital were divided in to three equal-sized treatment groups. In Group A, patients were treated with non-Gamma irradiated Beri honey. In Group B, Gamma irradiated Beri honey was used, and in Group C silver sulfadiazine was used. Treatment was for a period of four weeks. Pus swabs were taken at day zero and weeks one and four of treatment. Bacteria were identified using the analytical profile index system API 20E, 20NE and API Staph and antimicrobial susceptibility was done as per the Clinical and Laboratory Standards Institute 2010 guidelines. RESULTS: A total of 90 patients with wounds (n=90) took part in the trial. Out of 90 wounds, 47% were post-traumatic and 37% were postoperative. Overall, average length, width and depth of Group A patients' wounds were significantly reduced (p<0.0001). Out of 144 pus swabs; 99 and 45 were Gram-negative rods and Gram-positive cocci, respectively. Among these Pseudomonas aeruginosa (n=25) and Staphylococcus aureus (n=38) were the major pathogens. Interestingly, bacterial load gradually decreased from baseline to week four due to non-Gamma irradiated Beri honey. Moreover, both the Gram-negative rods and Gram-positive cocci displayed 100% resistance to commonly used antibiotics; the most effective drugs were carbapenem and vancomycin. CONCLUSION: Pakistani Beri honey could be used as an alternative therapeutic option for the management of infected wounds.

Role of Honey in Topical and Systemic Bacterial Infections.

BACKGROUND: The development of bacterial resistance to antibiotics has made it more difficult and expensive to treat infections. Honey is getting worldwide attention as a topical therapeutic agent for wound infections and potential future candidate for systemic infections. OBJECTIVES: The purpose of this review was to summarise different antibacterial bio-active compounds in honey, their synergistic interaction and their clinical implications in topical and systemic infections. In addition, contemporary testing methods for evaluating peroxide and non-peroxide antibacterial activity of honey were also critically appraised. DESIGN: MEDLINE, EMBASE, Cochrane Library, Pub Med, reference lists and databases were used to review the literature. RESULTS: Honey contains several unique antibacterial components. These components are believed to act on diverse bacterial targets, are broad spectrum, operate synergistically, prevent biofilm formation, and decrease production of virulence factors. Moreover, honey has the ability to block bacterial communication (quorum sensing), and therefore, it is unlikely that bacteria develop resistance against honey. Bacterial resistance against honey has not been documented so far. Unlike conventional antibiotics, honey only targets pathogenic bacteria without disturbing the growth of normal gastrointestinal flora when taken orally. It also contains prebiotics, probiotics, and zinc and enhances the growth of beneficial gut flora. The presence of such plethora of antibacterial properties in one product makes it a promising candidate not only in wound infections but also in systemic and particularly for gastrointestinal infections. Agar diffusion assay, being used for evaluating antibacterial activity of honey, is not the most appropriate and sensitive assay as it only detects non-peroxide activity when present at a higher level. Therefore, there is a need to develop more sensitive techniques that may be capable of detecting and evaluating different important components in honey as well as their synergistic interaction. CONCLUSIONS: Keeping in view the current guidelines for treatment of diarrhea, honey is considered one of the potential candidates for treatment of diarrhea because it contains a natural combination of probiotics, prebiotics, and zinc. Therefore, it would be worthwhile if such a combination is tested in RCTs for treatment of diarrhea.

In-vitro susceptibility of methicillin-resistant Stayphylococcus aureus to honey.

Wound infections caused by methicillin-resistant Staphylococcus aureus (MRSA) is becoming much complicated and costly to treat as antimicrobial resistance is quite common. Twenty five MRSA strains isolated from infected wounds and three ATCC reference strains were evaluated for their susceptibility to locally produced black seed (Nigella sativa), beri (ZiziphusJujuba) and shain honey (Plectranthus rugosus wall) by agar incorporation assay. Medically graded manuka honey (UMF 21+) was included as control. Locally produced black seed honey inhibited all clinical isolates at mean MIC of 5.5% (v/v), whereas manuka honey at mean MIC of 4.4% (v/v). The other two locally produced honey; beri and shain honey inhibited these isolates at 6.4% and 10.4% (v/v) respectively. The result of the study has demonstrated that indigenous black seed honey has comparable antibacterial activity to manuka honey and thus offers a good new addition to the existing honey resource for the treatment of wound infections.

A Randomized, Controlled Clinical Trial of Honey-Impregnated Dressing for Treating Diabetic Foot Ulcer.

OBJECTIVE: To investigate the effect of Beri-honey-impregnated dressing on diabetic foot ulcer and compare it with normal saline dressing. STUDY DESIGN: A randomized, controlled trial. PLACE AND DURATION OF STUDY: Sughra Shafi Medical Complex, Narowal, Pakistan and Bhatti International Trust (BIT) Hospital, Affiliated with Central Park Medical College, Lahore, from February 2006 to February 2010. METHODOLOGY: Patients with Wagner's grade 1 and 2 ulcers were enrolled. Those patients were divided in two groups; group A (n=179) treated with honey dressing and group B (n=169) treated with normal saline dressing. Outcome measures were calculated in terms of proportion of wounds completely healed (primary outcome), wound healing time, and deterioration of wounds. Patients were followed-up for a maximum of 120 days. RESULTS: One hundred and thirty six wounds (75.97%) out of 179 were completely healed with honey dressing and 97 (57.39%) out of 169 wtih saline dressing (p=0.001). The median wound healing time was 18.00 (6 - 120) days (Median with IQR) in group A and 29.00 (7 - 120) days (Median with IQR) in group B (p < 0.001). CONCLUSION: The present results showed that honey is an effective dressing agent instead of conventional dressings, in treating patients of diabetic foot ulcer.

Evaluation of the antibacterial activity of selected Pakistani honeys against multi-drug resistant Salmonella typhi.

BACKGROUND: The development of resistance to conventional anti-typhoid drugs and the recent emergence of fluoroquinolone resistance have made it very difficult and expensive to treat typhoid fever. As the therapeutic strategies become even more limited, it is imperative to investigate non-conventional modalities. In this context, honey is a potential candidate for combating antimicrobial resistance because it contains a broad repertoire of antibacterial compounds which act synergistically at multiple sites, thus making it less likely that the bacteria will become resistant. The in vitro antibacterial activity of 100 unifloral honey samples against a blood culture isolate of multi-drug resistant (MDR) Salmonella typhi were investigated. METHODS: All honey samples were evaluated for both total (acidity, osmolarity, hydrogen peroxide and non-peroxide activity) and plant derived non-peroxide antibacterial activity by agar well diffusion assay at 50% and 25% dilution in sterile distilled water and 25% in catalase solution. Manuka (Unique Manuka Factor-21) honey was used for comparison. The phenol equivalence of each honey sample from 2% to 7% (w/v) phenol was obtained from regression analysis. The antibacterial potential of each honey sample was expressed as its equivalent phenol concentration. The honey samples which showed antibacterial activity equivalent to or greater than manuka honey were considered therapeutically active honeys. RESULTS: Nineteen honey samples (19%) displayed higher hydrogen peroxide related antibacterial activity (16-20% phenol), which is more than that of manuka honey (21-UMF). A total of 30% of the honey samples demonstrated antibacterial activity between 11 and 15% phenol similar to that of manuka honey while 51% of the honey samples did not exhibit any zone of inhibition against MDR-S. typhi at 50% (w/v) dilution. None of the indigenous honey samples displayed non-peroxide antibacterial activity. Only manuka honey showed non-peroxide antibacterial activity at 25% dilution (w/v) in catalase solution. CONCLUSIONS: The honey samples which displayed antibacterial activity equal to or greater than manuka honey may be useful in the clinical conditions where higher hydrogen peroxide related antibacterial activity is required. Manuka honey, which is known to possess non-peroxide antibacterial activity, warrants further evaluation in a suitable typhoid animal model.

Histopathological findings in goiter: A review of 624 thyroidectomies.

OBJECTIVES: To identify the histopathological patterns of goiter in thyroidectomy specimens and their frequency in relation to age and gender of the patients. METHODOLOGY: We present a retrospective data of 624 thyroidectomy specimens diagnosed over a period of six year (2007-2012) at the Department of Pathology, Holy Family Hospital, Rawalpindi, Pakistan. RESULTS: A total of 624 consecutive thyroidectomy specimens were selected. Patient's age ranges from 11-89 years, 541 females and 83 males. There were 512 (82%) non-neoplastic lesions, which includes; 475 (76.1%) multi-nodular goiter (MNG), 16 (2.6%) Hashimoto thyroiditis, 11 (1.8%) colloid goiter, 4 (0.6%) toxic goiter, 2 (0.3%) chronic lymphocytic thyroiditis, 2 (0.3%) tuberculous thyroiditis and 2 (0.3%) miscellaneous. From 112 (18%) neoplastic lesions, 43 (6.9%) were adenomas (41 females and 2 males) and 69 (11.0%) were carcinomas (58 females and 11 males). Peak age for thyroid malignancy was 3rd to 4th decades. The histological subtypes of thyroid carcinomas includes, 35 (5.6%) follicular variant of papillary carcinoma (FVPC), 15 (2.5%) well-differentiated tumor of uncertain malignant potential (WDT-UMP), 6 (1%) medullary carcinomas, 6 (1%) papillary carcinomas, 3 (0.5%) anaplastic carcinomas, 2 (0.3%) follicular carcinomas and 2 (0.3%) other carcinomas. Twenty-nine (4.6%) neoplastic lesions were associated with MNG, includes; 2 (3.5%) follicular adenomas, 3 (0.5%) WDT-UMP and 4 (0.6%) FVPC. CONCLUSIONS: MNG is common and FVPC is the common thyroid cancer seen in females. The overall frequency of thyroid cancer is 11%. Follicular adenoma and FVPC appears to be associated with long standing MNG of iodine deficiency.

In vitro antibacterial activity of onion (Allium cepa) against clinical isolates of Vibrio cholerae.

BACKGROUND: Cholera is a major public health problem in developing countries of the world. Bacterial resistance, lack of surveillance data and proper microbiological facilities are major problems regarding diagnosis of cholera. The spread of microbial drug resistance is a global public health challenge that results in increased illness and death rate. Newer antimicrobials or agents are urgently required to overcome this problem. This work was therefore done to investigate the antimicrobial potential of onion against thirty-three clinical isolates of Vibrio cholerae. METHODS: The extract was prepared by reflux extraction method. Antibacterial screening of clinical isolates of V. cholerae was done by agar well diffusion method. Agar dilution method was used to assess the Minimum Inhibitory Concentration (MIC). RESULTS: All tested strains of V. cholerae were sensitive to onion (Allium cepa) extracts of two types (purple and yellow). Purple type of extract had MIC range of 19.2-21.6 mg/ml. The extract of yellow type onion had an MIC range of 66-68.4 mg/ml. CONCLUSION: The results indicated that onion (Allium cepa) has an inhibitory effect on V. cholerae. Keeping in view the anti-bacterial activity of this compound can be exploited as a therapeutic agent in an animal model. This finding is a positive point for further investigation of this herb of traditional medicine.