Vol. 42, September, 2015

1/42  MICROBIOLOGICAL AND MOLECULAR STUDY ON MICROORGANISMS RELEATED TO ACNE VULGARIS IN SOME EGYPTIAN HOSPITALS.

Moselhy S. Mansy, Kamel A. Al-Ghreib and Moaz E. Hassan

Microbiology and Immunology Department, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.

    This work was carried out on 284 patients with acne vulgaris attending the dermatology outpatient clinics of three hospitals in Egypt, during the period from December 2013 to February 2015. The cases of acne vulgaris were clinically diagnosed by the dermatologists. The samples were collected from the lesions of acne vulgaris using sterile stainless steel acne spoon, then using 2 sterile cotton swabs for each patient: One for direct Gram stained smear and microscopically examined, the other swab dipped in thioglycolate broth tube which were kept in ice box until transfer to microbiological laboratory. The thioglycolate broth was subcultured on suitable medium, then the growing microorganisms were identified by conventional methods. Staphylococcus epidermidis and Staphylococcus aureus, Malassezia fufur, Klebsiella pneumoniae and Propionibacterium acnes were isolated at frequencies 37 %, 20.4%, 12.3% 9.9% and 5.6% respectively. Identification of fifty S. epidermidis strains were confirmed by API staph system and ERIC-PCR. The API identification going hand in hand with the conventional method in 45 strains (90%) while PCR was going hand in hand in identification of  43 strains (86%).The biofilm production of isolated S. epidermidis and S. aureus (105 and 58 strains respectively) was done, we found that 10.5% of tested S. epidermidis and 13.8% of tested S. aureus produce strong biofilm. The isolated stains of microorganisms were tested for different antibiotics, S. epidermidis strains were sensitive to amikacin, ciprofloxacin and chloroamphenicol (93.3%, 81% and 78.1% respectively ), and were resistant to ampicillin, erytheromycin and tetracycline (92.4%, 56.2% and 54.3% respectively).We concluded that we should test the organisms isolated from acne vulgaris especially Gram-positive bacteria in chronic cases for biofilm production dute to bacterial biofilm may have a role in resistance to antibiotic repeatedly used in treatment. Also the production of biofilm may have a role in pathogenesis of acne vulgaris.

 

2/42  COMPARATIVE STUDIESONTHE ANTIMYCOTIC ACTIVITIES OF COMMERCIALLY AVAILABLE VAGINAL DOUCHES IN THE EGYPTIAN MARKET

Zahraa M. Mebad, Sameh M.M. Abdelghani*, Ahmed O. El-Gendy* and

Magdy A. Amin**

Microbiology and Immunology Department, Faculty of Pharmacy, Nahda University, Egypt.

*Microbiology and Immunology Department, Faculty of Pharmacy, Beni-Suef University, Egypt.

**Microbiology and Immunology Department and Biotechnology Centre, Faculty of Pharmacy, Cairo University, Egypt

Vaginal douches were the drug of choice for the treatment of vaginal candidiasis, for a period, but nowadays these vaginal douches become restricted in United States, although in our developing countries such as Egypt, still in use. Research articles show that women who use douches regularly have more health problems than women who do not. Vaginal flora represents one of the most important defense mechanisms, as in the healthy vagina, hydrogen peroxide (H2O2) &Lactobacilli protect against endogenous bacteria. So douching predisposes to disrupt these normal vaginal defense mechanisms. Sixty vaginal discharge samples were collected from patients experiencing itching and vaginitis. Clinical left over specimens have been recovered from different Beni-Suef Public Hospitals, Beni-Suef, Egypt. The identification was done using microscopical examination and Candi select-4 media. The comparative study was applied using agar diffusion method of commercially available douches in different concentrations against different Candida albicans. Then the effect of selective vaginal douches on various virulence factors were measured including biofilm formation, phospholipase, and proteinase activity on isolated Candida albicans. From the 60 vaginal discharge samples, only 47 have been identified as Candida albicans. In addition, by making sensitivity test for 10 commercially available vaginal douches through agar diffusion method we found that only three of them were active and the others with low or no activity. Interestingly, when studying 9oe98 Candida albicans, a dramatically increase in phenotypic expression of their virulence factors were observed. In conclusion, according to our study, it is recommended for women not to use douches in the course of treatment as these will harm by increasing the infectivity during candidiasis.

 

3/42  PHENOTYPIC AND GENOTYPIC CHARACTERIZATION OF DIFFERENT CLASSES OF β-LACTAMASES AMONGST ACINETOBACTER SPP. ISOLATED FROM EGYPTIAN HOSPITALS.

Kareem A. Sofy, Amal E. Saafan, Sameh M.M. AbdelGhani and Magdy A. Amin

Microbiology & Immunology Department, Faculty of Pharmacy, Beni-Suef University, Egypt.

*Microbiology & Immunology Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt.

Acinetobacter spp. resistance towards β-lactam antibiotics is mediated mainly by different classes of β-lactamases production; detection of some genes responsible for production β-lactamases is the objective of the study. One hundred fifty bacterial isolates were recovered from blood, sputum, and urine specimens from different hospitals in Egypt. Sixty-nine isolate were identified as Acinetobacter baumannii using traditional biochemical tests, CHROM agar and Micro Scan. Acinetobacter baumannii isolates were grouped into carbapenem resistant group (GP1), cefotaxime, ceftazidime and cefoxitin resistant group (GP2) and carbapenem, cephalosporin non-resistant group (GP3). Carbapenemase activity was screened using modified Hodge Test (MHT) for GP1.Metallo-β-lactamases screening was performed for MHT positive isolates using double disk synergy test (DDST) and combined disk test (CDT). Amp C activity was screened using Amp C disk test with Tris-EDTA, DDST and CDT for GP2. Finally, PCR amplification of blaoxa-51like, blaoxa-23like, blaIMP-like, blaVIM-like,andblaADC-like genes was performed for isolates that showed at least two positive results of three for both AmpC and carbapenemases phenotypic screening tests(obvious activity), in addition to GP3 (for comparison). Antibiogram of 69 pure Acinetobacter baumannii isolates resulted in 57, 64, and 2 isolates enrolled into GP1, GP2 and GP3 respectively. Carbapenemase activity was shown by 49(85.9%) isolate using MHT. Metallo-β-lactamases screening revealed 32(65.3%) and 35(71.4%) using DDST and CDT, respectively. AmpC activity was shown by 43(67.2%) and 50 (78.1%) isolates using AmpC disk test with Tris-EDTA, and both DDST and CDT, respectively. Twenty-seven isolates showed obvious activity, all of them (100%) were harboring blaoxa-51likeand blaADC-like genes, while blaoxa-23like, blaIMP-likeand blaVIM-like genes were harbored by 23(85.2%), 9 (33.%) and no isolate respectively. GP3 isolates showed only positive blaoxa-51likeand blaADC-like genes. It isn’t possible to correlate resistance with presence of blaoxa-51likeand blaADC-like genes. A blaoxa-23likegene played an important role in carbapenem resistance when compared by blaIMP-like and blaVIM-likegene.

 

4/42  CHARACTERIZATION OF MULTI DRUG RESISTANT AND BETA LACTAMASES PRODUCING PSEUDOMONAS AERUGINOSA ISOLATES FROM DIFFERENT CLINICAL SPECIMENS

Moselhy S. Mansy, Abdel-Nasser A. El-Moghazy, Rmadan A. Al-Domany* and Ibrahim M. Naguib

Microbiology & Immunology Department, Faculty of Pharmacy (boys), Al-Azhar University, Cairo, Egypt.

*Microbiology & Immunology Department, Faculty of Pharmacy, Kafr El-Sheikh University, Cairo, Egypt.

Multiple antibiotic resistances in bacterial populations are currently one of the greatest challenges to the effective management of infections. Constant bacteriological monitoring of pathogens in hospital in general and specialized units are necessary to provide accurate data on the prevalence and antibiotic resistance pattern of specific pathogens. The aim of the present study was to assess the prevalence and resistance pattern of P. aeruginosa in different Egyptian hospitals. Prevalence and antibiogram pattern of P. aeruginosa isolated from different clinical and environmental samples were characterized and identified using Gram-stain, conventional cultures, API 20E and antibiotic susceptibility then phenotypically tested by Modified Hodge test, EDTA disc synergy test, AmpC disc test and double disc diffusion synergy test. A total of 304 P. aeruginosa isolates were selected from 615 clinical specimens. The most isolates were from pus (29.6%) followed by urine (19.7%) and sputum (18.7%). The most active drugs against P. aeruginosa was carbapenems (imipenem and/or meropenem) followed by pipracillin-tazobactam combination. In the present study the frequency of multiple antibiotic resistant isolates were 103 (33.9%) which classified into multi drug resistant (MDR) 56 (18.4%), extensively drug resistant 29 (9.5%) (XDR) and pan drug resistant (PDR) 18 (6%). This study examined the prevalence of P. aeruginosa, and its susceptibility patterns to different antibiotics. The presence of MDR, XDR and PDR P. aeruginosa isolates may be attributed to β-lactamases production. So clinical isolates  phenotypically tested by modified Hodge test (MHT) for detection of carbapenemases, EDTA disc synergy test for detection of metallo-beta lactamases, AmpC disc test for detection of AmpC β-lactamase and double disc diffusion synergy test for detection of extended spectrum β-lactamase (ESBLs). The rate of potential carbapenemases, metallo-beta lactamases, AmpC and ESBLs production were 65/95 (68.4%), 75/95 (78.9%), 12/20 (60%) and 0/19 (0%) respectively.

 

5/42  "REVIEW ARTICLE"

BACTERIAL BIOFILM AS   A MAJOR   CHALLENGE IN TREATMENT OF HOSPITAL ACQUIRED INFECTIONS

Ahmed Eid Alharbi

Microbiology and Immunology Department, Faculty of Medicine, Taibah University, Kingdom Saudi Arabia

Bacteria can live in a free environment, or in biofilms. Biofilms are interdependent communities of microorganisms that grow on living or inert surfaces and have the ability to surround themselves with polymers. They are very organized and cooperative groups. Bacteria inside biofilms are commonly resistant to   most antibiotics. The transplanted devices contribute in the formation of biofilm. One of the biggest challenges facing doctors is the   lack of approach for the treatment of infections without removing the implanted device. Biofilms usually associated with health care infection and causes chronic polymer associated with the medical device implanted, such as a catheter inside the blood vessels which, and urinary catheter.58.95% of urinary tract infections because of the catheter, and 87% of cases of bloodstream infections arise from indwelling vascular catheter. It was also noted that bacterial biofilms that formed in the prosthetic joint are resistance for all antibiotics. The use of antibiofilm-coated medical devices reduces the chance of bacterial adhesion and growth on the surface of an implanted device. An example of antibiofilm materials are the small molecules and enzymes and more anti-adhesion coating are under development approach for the treatment and prevention of infection due to bacterial biofilms.

 

6/42  ROLE OF BIO AND PHOSPHOROUS FERTILIZATION IN INCREASING GROWTH AND PRODUCTIVITY OF CANOLA UNDER NEW VALLEY CONDITIONS

Mona M. El-shazly

Soil Fertility and Microbiology Department, Desrt Research Center

To evaluate the effect of biofertilization, i.e. Azotobacter chroococcum and Mycorrhizae (Glomus macrocarpium) were added with three Phosphate levels (50, 75 and 100 kg/f P2O5) on the yield and the components of canola plants (Serw-4),i.e. amino and fatty acids  content, as well as the on microbial activities in soil. A field experiment was conducted at El kargha Research Station, New Valley during two successive seasons. The obtained results showed the positive response of canola to different biofertilization treatments and phosphate levels for most of the studied parameter. Also results indicated that plant inoculation with mixed treatment of A. chroococcum + Glomus macrocarpium and third P level showed the highest increases of both seed yield/plant and oil yield/in both seasons, as compared with the un-inoculated plants. Addition of biofertilizers significantly improved microbial activities in the rhizosphere area of canola plants, represented by Nitrogenase and Phosphatase enzyme, Total microbial counts, Azotobacter densities and root mycorrhizal colonization %.

 

7/42  CHARACTERIZATION OF ACINETOBACTER BAUMANNII CLINICAL ISOLATES FROM KING ABDULAZIZ UNIVERSITY HOSPITAL IN JEDDAH, SAUDI ARABIA

Nayera A. Moneib, Heba M. Amin**, Aymen S. Yassin*, Dalia El-Hossary***and Asif Jiman-Fatani****

Department of Natural Products and Alternative Medicine, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia

*Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt

**Department of Microbiology, Faculty of Pharmacy, Modern Sciences and Arts University (MSA), 6th of October, Giza, Egypt

***Department of Medical Microbiology and Immunology, Faculty of Medicine, Zagazig University, Zagazig, Egypt

****Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia

Acinetobacter baumannii has emerged as one of the most common pathogens responsible for nosocomial infections, particularly in intensive care units. This study is carried out to characterize A. baumannii isolates among the patients admitted to King Abdulaziz University Hospital and find out their antimicrobial resistance and virulence patterns, in addition to the correlation with the underlying diseases.Clinical isolates of A. baumannii, collected from patients were properly identified using automated identification system. All isolates were subjected to proper antimicrobial susceptibility testing and minimum inhibitory concentration determination. All isolates were characterized by phenotypic testing for virulence factors. A total of 62 isolates were identified as A. baumannii. Most of the patients were undergoing some sort of surgical operation and were under total parenteral nutrition. Around 88.7 % of the isolates were multi drug resistant.  Most of the isolates were sensitive to colistin followed by tigecycline. The isolates showed varying degrees of biochemical activity when tested for gelatinae, lipase, haemagglutionation and biofilm formation.  There was an obvious increase in Acinetobacter infections among congestive heart failure and diabetic patients. A. baumannii remains one of the most common pathogenic microorganisms in intensive care units in hospitals. It still shows high patterns of resistance to most antimicrobials. The expression of virulence genes is variable among the different isolates.

 

8/42  IMPROVING THE PRODUCTIVITY OF SOME SUMMER FODDER CROPS USING DIFFERENT BIOFERTILIZER TECHNIQUES UNDER NEW VALLEY CONDITIONS

Mona M. El-Shazly

Soil Fertility and Microbiology Department, Desrt Research Center

To investigate the effect of different biofertilization techniques on productivity of some summer fodder crops guar (Cyamposis tetragonoloba L.) and Pearl millet (Pennisetum glaucum L.) under desert soil conditions. A field experiment was carried at the agriculture experimental station at El-Kharga Oasis, New Valley Governorate, Egypt. Biofertilization treatments (Azotobacter chroococcum, Bacillus circulans and Mixture) added to soil, foliar application of bacterial extracts, as foliar application, Micronutrients foliar application with biofertilizers or bacterial extracts. Obtained results indicated that, mixed biofertilization treatments with micronutrients foliar application  had the highest record for  both guar  and Pearl millet plant growth parameters, yield and its components as well as mineral contents of seeds (N,P,K as macronutrients) and (Zn, Mn, Fe and Cu as micronutrient). From the previous results it could be recommended using the mixture of biofertilizer application to soil with micronutrients foliar application. These treatments increased guar and Pearl millet yield by 63 and 99 % increase over control respectively.

 

9/42  MYCOBACTERIAL HISTONE LIKE PROTEIN (HLP) HAS A SIGNIFICANT ROLE IN DNA FUNCTIONS

Shymaa Enany and  Sohkichi Matsumoto*

Department of Microbiology and Immunology, Faculty of Pharmacy, Suez Canal University, Egypt & *Division of Bacteriology, Department of Infectious Disease Control and International Medicine, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.

Histon like protein (Hlp) is known of possessing an intrinsic role in DNA metabolism and regulating the transcription of some genes in Mycobacterium species. We aimed to confirm its contribution in the regulation of the DNA related genes such as recA and dnaA by using hlp mutant Mycobacterium smegmatis. We succeeded to prove that Hlp could regulate the expression of recA and dnaA genes as we found up-regulation of recA and dnaA relative normalized expressions volume by the quantitative real time PCR in the hlp mutant strain by three fold higher than found in the wild strain. This finding suggests that Hlp controls DNA function and contributes to the pathogenicity in Mycobacterium species.

 

10/42  PHENOTYPIC AND GENOTYPIC METHODS FOR DETECTION OF SOME VIRULENCE FACTORS OF ENTEROCOCCI SPECIES ISOLATED FROM THE INTENSIVE CARE UNITS AT ASSIUT UNIVERSITY HOSPITALS

Hamada M.M. Halby and Nahla M. Elsherbiny*

Microbiology and Immunology Department, Faculty of Pharmacy, Al Azhar University Assiut Branch.

Microbiology and Immunology Department, Faculty of Medicine, Assiut University

Enterococci are one of the most common inhabitants of the gastrointestinal tract of humans. Enterococci have emerged as one of the leading cause of nosocomial infections, and recently have been persisting clinical problem due to their resistance to antibiotics and their virulence determinants that involved in the success of these microorganisms in the hospital setting. In this study, 7738 patients were admitted to 10 different ICUs at Assuit University Hospitals. Approximately 4% (285/7338) of those patients developed nosocomial infections (NI). Enterococci caused 10.5% (30/285) of these nosocomial infections. Fifty isolates (30 clinical isolates, 10 commensals isolates and 10 environmental isolates) were identified as enterococci. Isolates were diagnosed by culture, biochemical reactions and analytical profile index (API 20 strep). The production of certain virulence factors as cytolysin, gelatinase and biofilm were detected phenotypically and genotypically by PCR of (cylA, gelE, esp and vanA/B genes). Phenotypic detection of virulence factors revealed that 12%of the isolates were β hemolytic and 48% of the isolates were positive for gelatin hydrolysis. The ability of enterococci to produce biofilm was detected by microtiter plate method. For genotypic detection of virulence genes, 62% for gelE gene, 12% for esp gene, 12% for vanB gene and 2% for vanA gene. cylA gene was absent from all isolates. Conclusion: The commonest enterococci species found in the ICUs were E. gallinarum, E. casseliflavus and E. faecalis. Positive correlation between genotypic and phenotypic methods for detection of gelatinase and biofilm (P=0.001, 0.044 respectively). In addition, significant correlation was recorded between vancomycin resistance by agar screening method, and detection of vanB gene was (P=0.006). We conclude that enterococci especially vancomycin resistant caused a significant percentage of health care acquired infections, intestinal carriage and environmental contamination in the ICUs of Assiut University Hospitals.

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