Blastocystis spp., Cryptosporidium spp., and Entamoeba histolytica exhibit similar symptomatic and epidemiological patterns in healthcare-seeking patients in Karachi.

In this study, we collected data on the incidence of enteric parasites in healthcare-seeking individuals along with their symptoms to quantify the potential roles of factors such as age, sex, and seasonality in infection. We performed analysis to identify factors which could help differentiate parasitic infection from other causes of gastrointestinal illness in a community. The size of the patient population (n = 339), patient selection methodology, collection methods, and statistical analysis followed approaches from similar studies in core clinical journals. Ethical approval was obtained from the University of Karachi's Ethical Review Board. Fecal specimens (n = 339) submitted by symptomatic patients were collected from two clinical laboratories, along with information about the patients' age, sex, and symptoms. We found that symptoms of fever, vomiting, and constipation were 100 % predictive of finding a parasitic infection, while diarrhea was 88 % predictive of a parasitic infection. Gastrointestinal parasite-positive patients reported diarrhea (~60 %), vomiting (~30 %), fever (~25 %) and constipation (~25 %), while parasite-negative patients exhibited a symptomatic profile without fever, vomiting, and constipation. The distribution of symptoms in parasite-positive patients remained relatively invariant regardless of the parasite identified. Blastocystis spp.-mono-infected patients reported a similar profile to patients positive for Entamoeba histolytica/Entamoeba dispar and Cryptosporidium spp. Most parasitic infections exhibited a strong seasonal pattern, with a peak incidence in summer months. Infection by Blastocystis spp. was the most prevalent, and it was the only infection mathematically correlated to rainfall by Pearson's method. We observed no increase in healthcare-seeking behavior following a stressful community event, namely, the attempted assassination of Benazir Bhutto in Karachi. The data suggest that parasitological testing would produce a high yield of positive results when performed on healthcare-seeking patients in Karachi in 2007 with symptoms of fever, vomiting, or constipation and a low yield when performed on patients noting only abdominal pain. Parasitological testing also produces a higher yield on patients seen in summer months.

In vitro antifungal sensitivity of fluconazole, clotrimazole and nystatin against vaginal candidiasis in females of childbearing age.

BACKGROUND: Vaginal candidiasis is the most common infection of females. A large variety of antifungal drugs are used for treatment. The objective of this study was isolation and identification of Candida from high vaginal swabs and in vitro antifungal activity of Clotrimazole, Fluconazole and Nystatin against Candida. METHODS: Two hundred and fifty high vaginal swabs were collected from females reporting at different hospitals of Karachi. Wet mount was performed to observe the budding cells of Candida. Vaginal swabs were cultured on Sabouraud's dextrose agar with added antibiotics. Plates were incubated at room temperature for seven days. Chlamydospores of Candida albicans were identified on corn meal agar. Species of Candida were identified on Biggy agar. In vitro antifungal activity of Clotrimazole, Fluconazole and Nystatin was performed by MIC (Minimum inhibitory concentration), well diffusion method and disc diffusion method. RESULTS: Out of 250 high vaginal swabs, Candida species were isolated in 100 (40%) of cases. Out of 100, C. albican 30 (30%), C. tropicalis 21 (21%), C. parapsillosis 10 (10%), C. parakrusi 8 (8%), C. glabrata 8 (8%), C. krusei 3 (3%) were isolated. In vitro antifungal activity indicated Clotrimazole (MIC 16 and 8 microg/ml) effective against 68 (70%) of Candida SPP, Fluconazole (MIC 64 and 32 microg/ml) effective against 29 (36.2%) and Nystatin disc (100 units) was 51 (63.5%) effective. CONCLUSION: C. albicans was mainly isolated. Clotrimazole was more effective as compared to Fluconazole and Nystatin. Antifungal susceptibility testing should be determined before therapy to avoid treatment failures.

Antibacterial effect of NSAIDS on clinical isolates of urinary tract infection and diabetic foot infection.

Non Steroidal Anti-inflammatory Drugs (NSAIDS) are a group of chemically dissimilar agents that have primary effect of inhibition of prostaglandin's synthesis. Aspirin (Acetyl Salicylic Acid) is used as an analgesic, anti pyretic, anti-inflammatory agent and also have an anticoagulant effect. Tylenol (acetaminophen) is used as pain reliever. The objective of this study was to determine the effect of Aspirin and Tylenol against clinical isolates of urinary tract infection (UTI) and diabetic foot infections. A total of 100 clinical isolates were analyzed. Out of these 50 were urine samples from diabetic patients with UTI and 50 pus samples from diabetic foot infection. Bacteriological study was done by inoculating urine samples on Cysteine Lactose Electrolyte deficient (CLED) media. Pus samples were inoculated on Blood agar and MacConkey's agar. Identification was done by colony characteristics, gram staining and standard biochemical tests and Quick Test Strip (QTS-20) DESTO Laboratories, Karachi. Antibacterial effect of Aspirin and Tylenol were tested against 100 clinical isolates by Replica plate method, Agar well diffusion method and tube dilution method. Concentrations of Aspirin and Tylenol (10 microg, 50 microg, 100 microg, 500 microg, 1000 microg) were made in Muller Hinton media. Bacteria isolated from urine samples were Escherichia coli 30%, Staphylococcus aureus 20%, Enterococcus faecalis 10%, S. saprophyticus 10%, Proteus spp. 6%, Pseudomonas spp. 6%, S. pyogenes 6%, S. agalactiae 6%, S. epidermidis 4%, and Klebsiella spp. 2%. Bacteria isolated from pus samples were S. aureus 30%, Pseudomonas aeruginosa 18%, S. epidermidis 14%, Klebsiella pneumonia 12%, Proteus mirabilis 12%, E. coli 10%, P. vulgaris 4%. Aspirin was effective at 100-500 microg concentration against all isolates. Tylenol has marked effect on pathogens at 100 microg concentration. Aspirin and Tylenol along with analgesic, anti-pyretic, anti-inflammatory properties also have marked anti bacterial effect on isolates from UTI and Diabetic foot infections and inhibits the growth of both gram negative and gram positive bacteria, and both can be used synergistically with antibiotics for effective treatment.

Determination of antimicrobial effect of protamine by transmission electron microscopy and SDS PAGE on Pseudomonas aeruginosa isolates from diabetic foot infection.

OBJECTIVES: Diabetic foot infection is one of the major complications of diabetes leading to lower limb amputations. Isolation and identification of bacteria causing diabetic foot infection, determination of antibiotic resistance, antimicrobial potential of protamine by electron microscopy and SDS-PAGE analysis, arethe aims of this study. MATERIALS AND METHODS: 285 pus samples from diabetic foot infection patients were collected from different hospitals of Karachi and Capital Health Hospital, Halifax, Canada. Clinical history of each patient was recorded. Bacterial isolates were cultured on appropriate media; identification was done by morphology, cultural and biochemical tests. Effect of protamine against multi drug resistant strains of Pseudomona aeruginosa was checked by minimum inhibitory concentration in 96 well micro-titer plates. The isolates were grown in bactericidal concentration of protamine on plates to isolate mutants. Effect of protamine on protein expression was checked by SDS- PAGE and ultra-structural morphological changes by transmission electron microscopy. RESULTS: Results indicated prevalence of foot infection as 92% in diabetic patients. Major bacterial isolates were Staphylococcus aureus 65 (23%), P. aeruginosa 80 (28.1%), Klebsiella spp. 37 (13%), Proteus mirabilis 79 (27.7%), and Escherichia coli 24 (12%). These isolates were highly resistant to different antibiotics. MIC value of protamine was 500 µg/ml against P. aeruginosa. SDS-PAGE analysis revealed that protamine can suppress expression of various virulence proteins and electron micrographs indicated condensation of cytoplasm and accumulation of protamine in cytoplasm without damaging the cell membrane. CONCLUSION: P. aeruginosa and S. aureus were the major isolates expressing multi-drug resistance and protamine sulfate represented good antimicrobial potential.