Frequency of antibiotic associated diarrhea caused by Clostridium difficile among hospitalized patients in intensive care unit, Kerman, Iran.

AIM: This study evaluated the frequency of C. difficile and CDAD in the ICU of Shahid Bahonhar Hospital, Kerman, Iran. BACKGROUND: Clostridium difficile (C. difficile) is the most important antibiotic associated diarrhea agent in intensive care unit (ICU) patients. Based on its toxin producing ability, C .difficile is divided to toxigenic and non-toxigenic strains. METHODS: A total of 233 diarrheal samples were collected from ICU patients. The samples were cultured on Clostridium difficile medium with 5% defibrinated sheep blood containing cycloserine (500 mg/L), cefoxitin (16 mg/L) and lysozyme (5mg/L). The isolates were confirmed as C. difficile by polymerase chain reaction (PCR) of 16s rRNA gene and the presence of toxins genes (tcdA, tcdB, cdtA and cdtB) was also confirmed. Then, the toxin production of isolates was evaluated using ELISA. RESULTS: C. difficile was isolated from 49 (21%) out of 233 samples. The total isolates fell into the A-/B-/CDT- (48.97%), A+/B-/CDT- (28%), A+/B+/CDT- (20.4%) and A+/B+/CDT+ (2%) types. Both types of C.difficile, A-/B-/CDT- and A+/B-/CDT-, which account for 77.5% of all isolates, were unable to produce the toxin (nontoxigenic). On the other hand, A+/B+/CDT+ and A+/B+/CDT- (22.5%), were able to produce toxin or were toxigenic. CONCLUSION: The frequency of C. difficile was about 21% and only 22.4% of C. difficile isolates were able to produce toxins. It is expected that C. difficile A+/B+/CDT± are toxigenic and related to C. difficile associated diarrhea (CDAD). Additionally, about 4.7% of hospitalized patients in ICU suffered from CDAD, which is higher than the rates reported from industrialized countries. Notably, 28% of isolates were C. difficile A+/B-/CDT- which only carries tcdA genes without toxin production.

Prevalence of mip virulence gene and PCR-base sequence typing of Legionella pneumophila from cooling water systems of two cities in Iran.

Legionella pneumophila is the primary respiratory pathogen and mostly transmitted to human through water cooling systems and cause mild to severe pneumonia with high mortality rate especially in elderly both in hospitals and community. However, current Legionella risk assessments may be compromised by uncertainties in Legionella detection methods. Here, we investigated the presence of L. pneumophila mip gene in water samples collected from different hospitals cooling towers, nursing homes and building/hotels water coolants from two geographical locations of Iran (Kerman and Bam cities) during summer season of 2015 by both nested and real-time PCR methods. Analysis of the 128 water samples for presence of the mip gene by nested-PCR revealed, 18 (23%) positive cases in Kerman and 7(14%) in Bam. However, when samples were tested by real-time PCR, we identified 4 more new cases of L. pneumophila in the hospitals as well as nursing homes water systems that were missed by nested-PCR. The highest rate of contamination was detected in water obtained from hospitals cooling towers in both the cities (p≤0.05). Dendrogram analysis and clonal relationship by PCR-base sequence typing (SBT) of the L. pneumophila genomic DNAs in Kerman water samples showed close clonal similarities among the isolates, in contrast, isolates identified from Bam city demonstrated two fingerprint patterns. The clones from hospital water samples were more related to the L. pneumophila serogroup- 1.

Evaluation of the frequency of the IL-28 polymorphism (rs8099917) in patients with chronic hepatitis C using Zip nucleic acid probes, Kerman, Southeast of Iran.

Polymorphisms in the region of the interleukin IL-28 gene on chromosome 19 have been related with clearance of hepatitis C virus (HCV), a major human pathogen responsible for chronic hepatitis, cirrhosis and hepatocellular carcinoma. About 3% of the world's population is infected with HCV. The long-term response to therapy is influenced by many host and viral factors, and recent evidence has indicated that some host genetic polymorphisms related to IL-28 are the most powerful predictors of virological response in patients with HCV. This study assessed frequency of the IL-28 polymorphism (rs8099917) in 50 patients (39 men and 11 women ) with chronic hepatitis C using ZNA probe real time PCR new method . All patients were tested for genotype of HCV and the HCV viral load. In parallel, the levels of SGOT, SGPT and ALK enzymes were assessed. Treatment using Peg-interferon alpha with ribavirin was conducted for patients and subsequently samples were collected to detect any change in viral load or liver enzyme rates. The overall frequency of the TT allele is 74%, TG allele 20% and GG allele 6% and the percent of patients who had T allele was 84%. Clear reduction in viral load and liver enzymes was reported in patients with the T allele. Especially for genotype 1 which is relatively resistant to treatment, these alleles may have a role in this decline. In conclusion, we showed that IL-28 polymorphism rs8099917 strongly predicts virological response in HCV infection and that real-time PCR with Zip nucleic acid probes is a sensitive, specific and rapid detection method for detection of SNPs which will be essential for monitoring patients undergoing antiviral therapy.

Antiviral Activities of Honey, Royal Jelly, and Acyclovir Against HSV-1.

INTRODUCTION: Herpes simplex virus type 1 (HSV-1) belongs to the Herpesviridae family and genus simplex virus. This virus is usually acquired during childhood and is transmitted through direct mucocutaneous contact or droplet infection from infected secretions. The aim of the present study was to compare antiviral effects of honey, royal jelly, and acyclovir on herpes simplex virus-1 in an extra-somatic environment. MATERIALS AND METHODS: Vero cells were cultured in the Dulbecco's Modified Eagle's Medium (DMEM) along with 10% fetal bovine serum (FBS) in 12-welled microplates. Various dilutions of honey, royal jelly, and acyclovir (5, 10, 50, 100, 2500, 500, and 800 µg/mL) were added to the Vero cells along with a 100-virus concentration of TCID50. The plaque assay technique was used to evaluate the antiviral activities. RESULTS: The results showed that honey, royal jelly, and acyclovir have the highest inhibitory effects on HSV-1 at concentrations of 500, 250, and 100 µg/mL, respectively. In addition, honey, royal jelly, and acyclovir decreased the viral load from 70 795 to 43.3, 30, and 0 PFU/mL at a concentration of 100 µg/mL, respectively. CONCLUSION: The results of the present study showed that honey and royal jelly, which are natural products with no reports about their deleterious effect at least in laboratory conditions, can be considered alternatives to acyclovir in the treatment of herpetic lesions. However, it should be pointed out that further studies are necessary to substantiate their efficacy because hard evidence on their effectiveness is not available at present.

In vitro evaluation of anti-herpes simplex-1 activity of three standardized medicinal plants from Lamiaceae.

BACKGROUND: Rosmarinic acid (RA) is a phenolic acid with antioxidant and anti-viral effects. We have studied anti-herpes simplex virus type 1 (HSV-1) effect of three medicinal plants from Lamiaceae family which have been standardized on the basis of RA content. MATERIALS AND METHODS: Methanolic extract of Teucrium polium, Ziziphora clinopoides, and Salvia rhytidea was prepared by maceration method and RA content of the plants was determined using a spectrophotometric method. Maximum nontoxic concentration (MNTC) of the extracts was determined using neutral red method. Serial dilutions of extracts up to MNTC were examined on Vero cells for anti-HSV-1 effect by plaque assay in comparison to acyclovir as a positive control. RESULTS: Among the tested extracts, T. polium contained the highest percentage of RA (1.8%w/w) and exhibited the least toxicity (MNTC = 1000 µg/ml). The greatest anti-HSV-1 was shown by T. polium and Z. clinopoides extracts which exhibited both time and concentration-dependent plaque inhibition. CONCLUSION: Considering the low toxicity and significant anti-viral effect of T. polium extract, this plant would prove valuable as an active anti-viral drug.