Vol. 3, April, 1999.

1/3 THE USE OF ANTIBODY CAPTURE HAEMADHERENCE

TESTS AS AN AID IN DIAGNOSIS OF HEPATITIS A VIRUS

INFECTION IN SERUM AND SALIVA.

D.F.I. El Fouhil

Department of Microbiology, Faculty of Medicine for Girls, Al Azhar University,

Nasr City, Cairo, Egypt.

Serum and saliva specimens taken from 40 acute hepatitis A patients, 30 patients in the convalescent phase of hepatitis A virus (HAV) infection, 15 subjects with history of past HAV infection and 15 healthy control subjects with no history of previous exposure to HAV, were subjected to testing for imrmmoglobulin M antibody to HAV (IgM anti-HA V) and immunoglobulin G antibody to HAV (IgG anti-HAV) using enzyme linked immunosorbent assay (ELISA) test and haemadherence tests in the form of IgM and IgG antibody capture haemadherence tests. In antibody capture haemadherence. tests, HAV-H21 virus strain propagated in Vero cell culture was used as the source of haemagglutinin and human group O erythrocyles were used as indicator cells. Antibody capture haemadherence tests reactivities were compared with ELISA test results. It was found that the specificity of IgM antibody capture haemadherence test was 100% in both serum and saliva and its sensitivity was 98.4% in serum and 96.7% in saliva. The specificity of IgG antibody capture haemadherence test was also 100% in both serum and saliva while its sensitivity was 100% in serum and 93.4% in saliva. Antibody capture haemadherence tests were found to be simple, rapid and inexpensive tests, with high degree of specificity and adequate sensitivity in diagnosis of HAV infection and in assessment of immunity and susceptibility to HAV. Differences were minimal between results obtained in serum and saliva specimens. By comparing them, it was found that serum and saliva are both adequate specimens for antibody capture haemadherence tests as well as for ELISA test. Because of the non invasive nature of sampling, saliva was considered as a satisfactory alternative to serum in diagnosis of HAV infection and in determination of the immune status to HAV by both techniques.

2/3 EFFECT OF MUSCULAR EXERCISE ON BLOOD LACTATE AND LACTATE DEHYDROGENASE ENZYME ACTIVITY DURING DIFFERENT PHASES OF MENSTRUAL CYCLE

N.E. Hashim, H.A. Abd El Sarnie and S.A. Kandil

Department of Physiology, Faculty of Medicine for Girls, Al Azhar University

The purpose of this study was to determine the e4ect of exhausted muscular exercise on blood lactate and serum lactate dehydrogenase (LDH) concentrations during the different phases of menstrual cycle. In the present work, t\venty fit female volunteers of average standard arthropomefric measures and with normal menstrual history were selected from the second year ofZagazig Faculty of Physical Education. While fasting, the volunteers were allowed to perform a physical exercise effort consisted of a progressive exercise test to exhaustion on a cycle ergometer. Blood samples were analyzed for lactate level and LDH activity in the destructive, proliferative and secretory phases of menstrual cycle. Our findings showed that blood exhausted exercise increased significantly lactate level in the three phases of menstrual cyck and serum LDH activity in both destructive and secretory phases only. In the pre-exercise period, blood lactate showed no significant difference between the three phases of menstrual cycle while serum LDH showed a significant decrease in the deatnistive phase versus the proliferative phase and significant increase in the proliferative phase versus the secretory phase and in the secretory phase versus destructive phase.. In post-exercise period, blood lactate showed a significant increase in the destructive phase versus the proliferative one and a significant decrease in the secretory versus the destructive phase while 'LDH showed 15.1 opposite effect with significant decrease in the destructive phase but significant Increase in the secretory phase. In conclusion we can say that the blood lactate and LDH activity play a role in the alteration ojfemale performance during the different phases of menstrual cycle.

3/3 TRACE ELEMENTS ALTERATIONS IN SOME RECURRENT AND ACUTE RESPIRATORY TRACT INFECTIONS IN CHILDREN

A. Abdel-Lateef, A. T. Mouawad and A.A. Gad

Health Research Department, National Center for Radiation Research and Technology

This study was conducted on 70 children suffering from recurrent tonsillitis, Tecwrent otiti: media and acute lower respiratory tract infections (ALRi). Patients hod a mean age of 4.67 ± LI years range 1-6.5years, 25 males and 25females. Tweiwy healthv children, age and sex matched with the patients, served as controls. Paiievits as well as controls were subjected to careful history taking; thoroz4gh clinical examination and laboratory investigations when needed to support the diagnosis. During the acute phase of infection blood samples were collected to demonstrate serum levels of zinc (Zn), Copper (Cu) and iron (Fe). The results showed that the patients had significantly lower levels of Zn and Fe when compared to the control group. The group ofpatients with ALRI had the lowest level ofzn, while the. recurrent otitis media group had the lowest level of iron. As regards the serum Cu, it tends to increase in patients with significant results in ALRJ and recurrent tonsillitis. The changes in trace elements status observed in this study may be due to the combined effects of infection and malnutrition.

4/3 PURIFICATION AND CHARACTERIZATION OF SM PECTINASE BIOSYNTHESIZED UNDER SOLID STATE FERMENTATION BY ASP. NIGER S-48TAT FROM THE WASTE PEELS OF SOLANUMMELANOGENUM (SM) COLLECTED FROM KITCHENS OF RESTURANTS IN CAIRO

M.S. Ammar, S.S'. Louboudy, M.S. Azab and M.M. Afifi.

Bot: and Microbiol. Dept, Fac. Sci., AI-Azhar Univ., Cairo, Egypt.

Purrfication and characterization of SM-pectinase produced under 5SF by A. niger, S-48TAT allowed to attack the waste peels of Sotanum melanogenum (SM) collected from the garbage of the kitchens of resturants in Cairo city has been undertaken. Purification process included the application of Ammonium Sulfate precipitation technique, dialysis and passing through afermentation gel ofsephadex G-200 colum. This readied in having a specific activity of837.2 units/mgprotein/ml corresponding to purification folds of 3.41 times. Data recorded for the properties of the purified eiuym in its reaction mixtures are: incubation time, 18 h. at room temperature; optimum pH, 4; optimum pectin concentration, 0.1%; thermal stability, 6(fC anda noticeable continuous increase in enzyme activity due to the incrrease of enzyme concentration. The enzyme was stable within pH (2.6-3) using citrate-phosphate buffer. Application of the introduced PCZ (pectin clearing zone) technique in thefield qfpectin-containing fruit juices in industry comparable to Novo-Pectmex enzyme proved the efficiency and precision of this technique from one hand and enabled the evaluation of SM-pectinase on the other hand. Therefore, a suggestion was given for the possible application of SM-pectinase in fruit juices industry.

5/3 SYMPTOMATIC RESPONSE FOLLOWING ONE WEEK TRIPLE THERAPY IN HELICOBACTER PYLORI INFECTED

PATIENTS

A. A.A. Farghaly, M.F. Rabea* and M.F. Mohamed*

Surgury Department, Al-Menofia University. * Microbiology department, Al-Azhar University.

Eighty eight patients from the surgical outpatient clinic ofMenofia University Hospital were complaining of dyspepsia. The isolation rate ofH.pylori by more than two of urease rapid test, culture, histology and the presence of IgG against H.pylori by ELISA technique was 86.2%. (24.1% in duodenal ulcer, 23% in gastritis and 21.8% in gastric ulcers). They were subjected to questionnaire to score their dyspepsia their symptoms were mostly heartburn 69.7%, epigastric pain 55.2%, dyspepsia (pain or discomfort, heartburn, nausea or vomiting) 46.1%, nausea 44.8%, acid regurge 16.8% and vomiting 3.4%. H.pylori was isolated from patients who had gastritis, duodenal and gastric ulcers in the following frequency 29.5%, 2 5% and 21.8%. The patients then were randomized into 3 groups. Group 0AM (30 patients were given omeprazole 20 mg, amoxycillin 1 gm and metronidazole 400 mg twice daily, group OCA (28 patients) were given omeprazole 20 mg, amoxycillin 1 gm and clarithromycin 250 mg twice daily, and group OCM (30 patients) were given omeprazole 20 mg, clarithromycin 250 mg and metronidazole 400 mg twice daily. Helicobacter pylori was eradicated in 89.3% of OCA group and 83.3% of OCM group and the least eradication was among OAM group which is 70%. The resolution of symptoms after eradication of H.pylori showed unconvencing results.

6/3 PATHOPHYSIOLOGICAL CHANGES IN SOME HAEMOSTATIC PARAMETERS IN EGYPTIAN BILHARZIASIS

M.M. El-Bayoumy, H.A. Abd El Sarnie, H.A. Abd El Hafiez* and M.A. Sorour **

Departments of Physiology, Internal Medicine * and Clinical Pathology**, Faculties

of Medicine, Al-Azhar University

Bilharziasis has been considered as an Egyptian AIDS and commonly has been associated with disseminated intravascular coagulation. This study included forty five patients suffering from bilharzial infestation and twenty healthy individuals with the same age and used as a control. Bilharzial patients were subdivided into two groups: the first one was compensated bilharziasis i.e. patient with hepatosplenomegaly "twenty cases", the second group was decompensated "twenty five patients" suffering from hepatic cirrhosis and ascites. All subjects were submitted to clinical history, examination and laboratory investigations for liver function tests (alkaline phosphatase (ALP), alanine aminotransferase (ALT) aspartate aminotransf erase, (AST) and gamma glutamyl transferase (GGT)), as a routine to prove infestation . Serum was analysed for determination of fibrinogen and plasminogen concentrations, tissue plasminogen activator (TPA), plasminogen activator inhibitor-l (PAI-l), fibrinogen degradation product (FbDP), D-dimer (D-D) and activated partial thromboplastin time (APTT). The results showed a progressive decrease in fibrinogen and plasminogen concentrations together with reduction of plasminogen activator inhibitor-l, while tissue plasminogen activator, fibrinogen degradation products and D-dimer showed a progressive increase parallel with the progress of the disease. Activated partial thromboplastin time showed significant prolongation only in decompensated group. It is concluded that bleeding tendencies in bilharzial patients are primary due to the defect in jlbrinolytic mechanism. Also, the amount of plasminogen activator inhibitor, available to bind and neutralize circulating tissue plasminogen activator, may be a critical factor in the progress of hyperfibrinolysis observed in Egyptian bilharziasis.

7/3 HISTOLOGICAL STUD- ON THE ENTEROTOXIGENICITY OF NON-IRRADIATED AND IRRADIATED BACILLUS CEREUS

M.N. F.afez and R.N El-Hifliawi

Department of Radiation Biology and Department of Drug Radiation Research, National Centre for Radiation Research and Technology -CRRT), P.O.Box 29, Nasr

City, Cairo, Egypt.

Evidence is pr6vided that an enterotoxin synthesL-ed during exponential growth by Bacillus cereus is responsible fbr the diarrheal-type and vomiting-type syndromes. These syndromes have been studied in vivo by investigating the histological changes of both the small intestine and stomach of the albino rats infected with non-irradiated and irradiated Bacillus cereus (NRRL 569).

8/3 PRODUCTION, PURIFICATION ANDCHARACTERIZATION OF EXTRACELLULAR FIBRINOLYTICENZYMES FROM

ASPERGILLUS FLA VUS

S.T. El-Sayed and *A.L. Kansoh

Biochemistry and *Microbial Chemistry Department, National Research Center,

Tahrir St, Dokki, Cairo, Egypt.

Fibrinolytic enzymes are produced extracellular adoptively in liquid culture of Aspergillus flavus containing either 1.3 % fibrin or 1.0% casein as a nitrogen source. The optimal initial pH value for high yield offibrinolytic enzyme production was 6.0 at 37°C for 9 6hr's incubation period. Three fibrinolytic isozymes F;, ft and Fj were produced and purified from Aspergillus flavus with specific activity 297.5, 327.4 and 40.3 U/mg protein respectively. The purification involved ammonium snlfate precipitation (50-80% saturation), DEAE-cellulose, Sephadex G-120 and Sephadex G-200 columns chromatography. Each enzyme was shown to be homogeneous chromatographically and disc electrophorically. The molecular weights ofFj, F? and F; were 49.0, 20.0 and 14.5 K Dal ton respectively, as estimated by gel filtration. Characterization of Fj and F2 with highest fibrinolytic activities were done. The pH optimum for the activity of Fi on fibrin was found to be 5.0 and for F2 was in the range of 5.0 to 8.0 at temperature 45 and 35 °C respectively. fi is stable in the pH range from 6.0 to 8.0, while F2 is stable in the pH 7.0 and8.0for 30min. Both isozymes are heat stable at 40 °Cfor 60 min. They have higher affinity towards buffalo fibrin than towards human fibrin with apparent Km values o/0.125 % and 0.34 %for Fiand F? respectively. Cu++ increased both fi andF2 activities by 8.0 and 17.0 %, while Zn* + caused marked inhibition by 33.0and 27.0 % respectively. Hg++and Mn++ caused complete inhibition.


9/3 SERUM MELATONIN, PROLACTIN, AND PROGESTERONE LEVELS IN WOMEN WITH PREMENSTRUAL DYSPHORIC DISORDER

H.M. Shalabi*, N. Abd-El-Ghaffar* and HAM. Gomaa**

*Obstet and Gynecol Dept. Faculty of Medicine for Girls and **Biochemistry Dept., Faculty of Pharmacy, Al-Azhar University.

The neuroendocrinological changes that occur in patients with Premenstrual Dysphoric Disorder (PMDD) are poorly understood. The mechanism is likely to be a complex interaction between gonadal hormones and neurotransmitters. Melatonin is the predominant pineal neurohormone. The gonadal changes associated with melatonin are mediated via the hypothalamus. The purpose of this study was to evaluate melatonin concentrations in patients with PMDD. Also serum prolactin (PRL) and progesterone levels were estimated for any correlation with melatonin secretion. During the symptomatic late lutealphase, serum melatonin concentrations were determined by Radioimmunoassay (RIA) in 12 women with PMDD and 12 normal control women (NC). Also serum PRL and progesterone were estimated to all women by RIA. Melatonin concentration in patient with PMDD were significantly lower than in NC (mean 25.25 VS 79.25 P< 0.05). Serum PRL were significantly higher in PMDD subjects than in NC (mean 39.08 VS 20.17 P < 0.05). Serum progesterone levels were lower in PMDD than NC (mean 20.42 VS 33.58 P < 0.05). The results showed inverse correlation between melatonin and PRL levels (r = 0.931, P <. 0.00J) and direct correlation between melatonin and progesterone levels (r^ 0.894, p _ 0.001). No correlation was found between PRL and progesterone level. Findings of the study suggest that lowered melatonin levels in women with PMDD may be due to involvement of the pineal gland in the pathogenesis of this disorder.

10/3 COMPARATIVE ANTIMICROBIAL ACTIVITY AND POSTANTIBIOTIC EFFECT OF LY 333328 AND VANCOMYCIN

AGAINST PENICILLIN-RESISTANT STREPTOCOCCUS

PNEUMONIAE AND METHICILLIN-RESISTANT

STAPHYLOCOCCUS A UREUS

BY M.A. Ramadan

Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Kasr El-Aini Str., Cairo, Egypt

LY 333328 is a new semisynthetic glycopeptide active against vancomycin-resistant streptococci and other gram-positive bacteria. In this study, the bactericidal activity of LY 333328 was compared with that of vancomycin using the minimal bactericidal concentration (MBC), time-kill kinetics and post-antibiotic effect (PAE). The test organisms were 5 and 6 clinical strains of penicillin-resistant Streptococcus pneumoniae (PRSPN) and methicillin-resistant Staphylococcus aiireus (MRSA) respectively. The results indicated thatMBCs for LY 333328 against PRSPN were almost the same asMICs, while MBCs for vancomycin were 2 to 4 folds higher than its MICs. Killing kinetics for the tested strains showed the potent antipneumococcal activity of LY 333328. ft was bactericidal at eight times its MIC for four of the five strains of PRSPN after 4.0 h exposure and bactericidal for all five strains after 6.0 h at the same concentrations. By comparison, vancomycin was bactericidal only after 6.0 h. No viable cells were detected after an overnight incubation in all tested strains treated with 8 times MIC of LY 333328, while the bacterial growth was observed, however in 3 out of 5 (60%) of the tested exposed to vancomycin under the same conditions. Exposure of the tested strains to 4 MICs, LY 333328 showed a longer PAE period (2.0 - 2.9 h) and (1.7-2.1 h) against PRSPN and MRSA respectively. Vancomycin at the corresponding concentration produced shorter PAEs on PRSPN (1.4-2.0 h) and for MRSA (1.1 -1.5 h). LY 333328 represents an advance in the activity of glycopeptide class of antibacterial agents and improved activity against gram positive pathogens, particularly streptococci suggest a significant therapeutic potential for this compound.

11/3 A STUDY ON METABOLISM, OXYGEN UPTAKE ANDHYDROGEN PEROXIDE PRODUCTION OF ENERGYYIELDING SUBSTRATES BY MYCOPLASMA CANIS

BY

R. Abdel Megid FROM

Microbiology Department, Faculty of Pharmacy, Cairo University, Kasr EI-Emi,

Cairo, Egypt.

Oxygen uptake during the metabolism of various substrates by whole cells of Mycoplasms canis (M canis) and H202 accumulation together with the oxidation of NADH and L-a-glycerophosphate (GP), by cells lysed with Triton X-100 was determined for eight isolates of M canis from various sites in infected cattle. Oxidation of glycerol at low concentrations (not glucose), NAOHandGP by M. canis isolates was accompanied by the accumulation of large quantities ofH2O2. However, metabolism of glucose and glycerol was indicated by a reduction in the pH of the suspending medium (saline and catalase). Lysed cells oxidisedNADH andGP with the production of approximately 1.0 molH202/mlO2 taken up, indicating presence of a GP oxidase. The importance of H2O2 production as a factor in the pathogenicity ofM canis indicates that it might depend upon the availability of glycerol in vivo.

12/3 VERIFICATION OF SOME NATURAL RECIPES AND PREPARATIONS FOR THE TREATMENT OF HAIR FALL

T.I. Khalifa*, O.D. El-Gindi**, H.E. El-Nazer*** and A.H. Ahmed**.

* Pharmacognosy Department, Faculty of Pharmacy, Al-Azhar University.

** Pharmacognosy Department, Faculty of Pharmacy, Al-Azhar University for Girls.

*** Pharmacology Department, National Research Centre, Dokky, Cairo.

Herbal preparations and recipes commonly used for the treatment of hair fall have been subjected to phytochemical screening and precise quality control analysis. This included physical characters, microscopical identification of the diagnostic elements of the herbal powdered mixture and examination of the different herbal extracts, as well as TLC investigation, GLC study of the volatile oils, saponifiable and unsaponifiable constituents. Quantitative estimation of the main active constituent was carried out. Biological activity of the different recipes was also done.

13/3 ENDOTHELIN ASSESSMENT AND DOPPLER

VELOCIMETRY OF UTERINE AND UMBLICAL VESSELS IN

CASES OF PREGNANCY ASSOCIATED HYPERTENSION

Y. Wafa**, E. El-Gindi*, G.A Serour**, and S. Abdel-Hamid*

* Department of Obstetrics and Gynecology, Faculty of Medicine, Minia University

**Department of Obstetrics and Gynecology, Faculty of Medicine, Al-Azhar University

Endothelin (ET-1) concentration in maternal blood is Found to be elevated in cases ofpre-eclampsia with negative correlation with the birth weight. The aim of this study was to demonstrate the correlation between (ET-1) level and the pathophysiolgical changes in uterine and umbilical blood flow and to find if there is correlation between (ET-1) and birth weight - Fourty cases with severe per-eclampsia and chronic hypertension with pregnancy were selected. They were given either lacidipine (Calcium Channel Blacker) or Labetalol (Beta adrenergic blocker). Ultrasonic fetal biometry and level of (ET-1) were done before and after treatment. It was found that (ET-1) was significantly higher in per-eclampsia than chronic hypertension cases. Also there was good correlation between (ET-1) andHead/ abdominal Circumference ratio. There was also positive correlation between (ET-1) and Systolic / Diastolic ratio of the studied a Blood vessels. It is concluded that (ET-1) is a good marker of pathophysiological changes in pre-elampsia and its level correlates with Doppler changes of umbilical and uterine arteries as well as head circumference / abdominal circumference ratio.

14/3 EFFECTS OF SOME QUINOLONES ON ADHESION, HAEMAGGLUTINATION, OUTER MEMBRANE PROTEINS

AND PLASMID PROFILE OF UROPATHOGENIC

ESCHERICHIA COLI

EM. El- Masry, A. A. Kadry, A. M. El- Sayed, and M. I. Hossainy

Department of Microbiology, Faculty of Pharmacy, Zagazig University

The ability of 62 E. coli isolates to adhere to human uroepithelial cells was studied. The adhesive ability of 62 E. coli isolates was found to be 90.3% with variable degrees of adherence. Four isolates were exhibited the highest adhesive ability were selected for this study. The effect of sub-inhibitory concentrations of tested quinolones ('/4 MICs) on adhesion ability ofE. coli to human uroepithelial cells was tested. No significant inhibition in adherence with nalidixic acid in three isolates, while one isolate showed enhancement of 'adherence. Norfloxacin had no significant inhibition in adherence on three isolates. On the other hand, there was significant decrease in adherence observed with ofloxacin, ciprofloxacin and pefloxacin on three isolates (P < 0.05). All tested E. coli isolates agglutinated human erythrocytes A in absence of D-mannose, whereas the haemagglutination ability of one isolate was altered in the presence of D-mannose. At sub-MICs, all tested quinolones with the exception of nalidixic acid inhibited the haemagglutination abilities of most isolates. Ofloxacin and ciprofloxacin affected the outer membrane proteins of the resistant isolate (No. 4), while, other quinolones had no effect. The effect of sub-MICs of tested quinolones on plasmid profile of E. coli isolate No. 4 was detected by agarose gel electrophoresis. All tested quinolones with the exception of nalidixic acid had plasmid curing effect. Pefloxacin was the most active curing agent compared to untreated cells.


15/3 MICROBIOLOGICAL, IMMUNOLOGICAL AND PCR ANALYSES FOR DISCRIMINATION OF SEPTIC ARTHRITIS

M.K. Okasha and H.A. Abdel-Salam

Department of Microbiology, Faculty of Pharmacy, Zagazig University,

Zagazig 44519, Egypt.

The serum and synovia! fluid concentration of interlukin-6 (IL-6) and Rose-Waaler (IgM-RF) correlate better than leukocytosis and C-Reactive protein (CRP) with septic arthritis. The level of IL-6 in the synovia/ fluid and serum of septic arthritis was found to be elevated up to 10 times more than aseptic one and'normal healthy. The concentration of IL-6 was found to be elevated (94.7% and 40%) in the synovial fluid (SF) and (39.5% and 0%) in the serum of septic and aseptic arthritis, respectively. The IL-6 level was significant, therefore it might be claimed to be used as a reproductive, rapid and specific diagnostic tool in septic arthritis and to differentiate between septic and aseptic arthritis. Meanwhile, CRP was elevated in the synovial fluid and serum of the septic and aseptic arthritis. However, the most prevalent autoantibodies found in rheumatoid arthritis (RA) were IgM-RF (70-80% positively) but in cases associated with systemic lupus erythmatosus were antinuclear factor (ANF) and anti-n-deoxyribonucleic acid (Anti-n-DNA) autoantibodies. Gram positive organisms appeared the most frequent isolates (63.3%), particularly S. aureus (30%) while gram negative bacteria represented only (31.6%) in septic arthritis. Moreover, Tubercle Bacilli, Bacteroides, Cryptococcus, Chlamydia and hepatitis virus were also found to be causative agents of septic arthritis. It was found that norfloxacin, ciprofloxacin, doxycycline, minocycline and rifampicin were effective against all isolated bacteria. The detection ofM. tuberculosis in the SF was carried out by poly me rase chain reaction (PCR) based on the amplification ofaDNA fragment of about 317 bp represents the insertion sequence-6110 (JS-611Q) of T.B. chromosome. Seven cases of pulmonary tuberculosis were analysed, 2 of these were SF positive for both PCR amplification and Ziehl-Neelsen slain (tuberculous arthritis). The PCR analysis ofHCVin the SF showed a DNA band of about 272 bp of the 5 *-untranslated region of HCV genome. Two out of 4 suspected cases were SF positive for both PCR analysis and immunoenzymatic assay (viral arthritis). This study showed, for the first time, the use of PCR amplification for the diagnosis of tuberculous and viral HCV arthritis. Therefore, the IL-6 level and PCR analysis were considered rapid, sensitive and specific as well as reproductive tests for discrimination and detection of septic arthritis and recommonded to be used in laboratoy diagnosis.

16/3 BIOSYNTHESIS OF PROTEASE (S) BY SOME SPECIES OF FUNGI UNDER THERMAL STRESS CONDITIONS

E.M. Mahmoud

Botany & Microbiology, Faculty of Science, Al-Azhar University

The biosynthesis of protease (s) enzyme by the thermophilic fungi, Sepedonium sp and Sporotrichum thermophtte, and a thermotolerant fungus, strain, Aspergillus fumigatus, has been considered. A spciefic medium was used as the basic production medium to which certain nutritional additives have been used in order to investigate their role in the biosynthetic productivies of protease(s) at 50 °C. Results indicates that the three strains failed to produce protease (s) in the absence of gelatin. Maximal production of the enzyme was obtained at two pH optima viz. 8 and 7.4 at 50 and 30 °C.. Both hist/dine and aspartic acids increase the yield up to the maximum for Sepedonium sp. and Sporotrichum thermophile, respectively. Generally supplementation of gelatin, starch and ammonium sulphate as well as calcium salts and magnasium sulphate salts exerted the highest level of protease (s) yield.

17/3 COMPARATIVE STUDY BETWEEN UREA (13C) BREATH TEST, DETECTION OF SPECIFIC SERUM ANTIBODIES AND CULTURE OF GASTRIC AND DUODENAL BIOPSIES FOR DIAGNOSIS OF HELICOBACTER PYLORI INFECTION

* M.Z. Al-Ghannam, ** T.R. El-Khamissy and *** M.W.S. Al-Din

* Clinical Pathology Department, Faculty of Medicine, Al-Azhar University,

(Assiut*) ** Microbiolgy Department, Faculty of Pharmacy, Al Azhar University,

(Assiut) and *** General Surgery Department, Faculty of Medicine,

Zagazig University

Urea (13C) breath test, detection of specific serum antibodies and culture of gastric and duodenal biopsies were used for diagnosis of Helicobacter pylori (H. pylori) infection. Twenty-five patients complaining of dyspepsia, active chronic gastritis, gastric or duodenal ulcer, but not related to alcohol or non steroidal anti-inflammatory drug-intake admitted to Zagazig University Hospital were screened for Helicobacter pylori infection. They were examnined and tested for diagnosis of infection and then after complete course of specific treatment, they were .examinedfor eradication of the infection. It was found that urea (13C) breath test and culture of gastric and duodenal biopsy for isolating the organism were positive in all cases (100%), while detection of specifc serum antibodies against H. pylori was only positive in 23 out of 25 (92%) cases and negative in 2 cases (8%). On the other hand and after complete drug regimen, urea (13C) breath test and biopsy culture were negative for 23 cases (92%) demonstrating relieve of infection, but still positive for 2 cases (8%). Detection of specific serum antibodies of the organism was still positive in 23 cases (92%) and negative in 2 cases (8%) after therapeutic treatment. The study indicated that urea (13C) breath test and culture of gastric and duodenal biopsies were more reliable for diagnosis and follow-up of treatment of H. pylori infection. This study also, suggests that urea (13C) breath test, the noninvasive technique is more rapid, accurate and easy done test. So, it is a more reliable method for diagnosis of either infection or relief of patients with H.pylori infection.

18/3 MICROBIOLOGICAL PROFILE OF CHRONIC DIARRHEA IN ADULTS

* M.O. Abbas, *S.M. Zaky, **K.M. Maklad, ***A.N. Haseeb, ***A.E. Abdel Ohany

* Microbiology and Immunology Department ** Parasitology Department*** Tropical Medicine Department, Faculty of Medicine, Am Shams University

This work was planned to study the microbiological andparasitological profile of stool samples from adult patients with chronic diarrhea. The study was performed on 25 patients and 10 healthv controls with matched age and sex. Each patient was exposed to thorough clinical examination and afiLil history taking. Stool samples were aseptically collected from patients and controls, cultured aerobically and anaerobically for bacterial pathogens, for detection of Rota virus by latex agglutination test and exam ined paraskologically for parasitic pathogens It had been found that most cases and control stool samples contained E.coli spp. (76% and 100% respectively). Two E.coli spp. Isolated from patients were found to be enteropathogenic strains (0124 and 0111). Some aerobic and anaerobic organisms were isolated only from patient's stool including Kiebsiella ozaenae (8%), Klebsiella pneumoniae (16%), Salmonella para typhi A (16%), Peptostreptococcus (12%), bacteroides corrodens (4%) and Bacteroides ovatus (4%), showing statistically significant difference between patients and controls. Fungi, Candida albicans, Aspergillus niger and Aspergillus fumigatus were isolated from patients (48%, 5% and 8% respectively) while only Candida albicans and Aspergillus niger were isolated from controls (30% both) Parasitological study ofstool samples revealed the presence of Entamoeba histolytica cysts in 4 cases (16%) and one control (10%); while Cryptospridium parvum was found in S patients (20%). Detection of Rota virus in stool samples showed statistically insignificant difference between patients and controls. Patients with history of Schistosomiasis or diabetes mellitus showed multiple isolates including bacteria, fungi and parasites. Duration of diarrhea and fever had insignificant correlation with types and numbers of isolates.

19/3 MICROBIOLOGICAL QUALITY OF SOME MEAT PRODUCTS IN LOCAL MARKET

G.A.A. El-Sharnouby and H.H. Abd El-Dayem

Food Sci. and Techn. Dept, Fac. of Agric., Al-Azhar Univ., Nasr City, Cairo, Egypt.

A total of 125 random samplesof meat products including 25 samples of each frozen minced meat, fresh sausage, cookedshawarma, luncheon.and Basterma were collected from Cairo city markets and examined for microbiological quality. Counts of several spoilage and pathogenic microorganisms were assessed. Besides, incidence and prevalence of four different food-poisoning bacteria were carried out. The total aerobic counts varies within the different samples. The highest count was attained 880 x 10" cells/gm in minced meat. Also, the highest numbers of psychrophilic bacteria, molds and yeasts, proleolytic and lipolytic bacteria were found in minced meat samples at counts 605 x 103 cells/gm, 50 x JO2 CFU/gm, 36x JO2 and 33 x 10' cells/gm, respectively. The pathogenic bacteria Streptococcus faecalis and Enteropathogenic E.coli were found in all type of samples at different prevalence ranged from 12% to 20% and 8% to 32% of samples, respectively. Poisoning bacteria, coagulase positive Staphylococcus aureus, Salmonella spp. and Clostridium perfringens were detected in some samples of minced meat, sausage and Sha\varma at different prevalence ranged from 16% to 32%, 4% to 12% and 4% to 8%, respectively. However, Listeria monocytogenes bacteria was detected only in two samples of minced meat. Furthermore, Public health hazard of the contaminated microorganisms was discussed.


20/3 BIOLOGICAL PROPERTIES OF SOME OPPORTUNISTIC

PATHOGENS IN WATER : PSEUDOMONAS AERUGINASA,

AEROMONAS SPP. AND STAPHYLOCOCCUS AUREUS

N.A. Hassanean* , P.M. Selim** , L.I. El - Seadawy** ,

H.T. El-Zanfly*, and E.M. Hassan**

*Water Pollution Control Department, National Research Center. '"Microbiology Department, Faculty of Medicine, Al Azhar University (Girls).

In this study 71 opportunistic pathogens isolated from different hospital water samples [32 Pseudomonas aeruginasa (P. aeruginosa), 23 Aeromonas spp. and 16 Staphylococcus aureus (S. aureus)] were tested for their antimicrobial susceptibility to ten commonly used antimicrobials. A survival experiment was done using one isolate of each of P. aeruginosa, Aeromonas hydrophila (A. hydrophila), Aeromonas sobria (A. sobria), Aeromonas caviae (A caviae) and S. aureus held separatly in water. E. coli (ATCC1422) and Streptococcus faecium (S. faecium) (ATCC 6057) were used as bacterial indicators. Water samples were withdrawn hourly for the first five hours, daily for a week and weekly for 3 weeks. The survival of the tested organisms was determined by changes in viable count against time. The disinfectant effect of ultra-violet (UV) radiation on 3 P. aeruginosa isolates , 3 Aeromonas isolates and 2 S. aureus isolates was studied by exposing them to Ul7 doses; 4 mJ / cm2, 8 mJ/cm2 and 12mJ/cm2 and the bacteria were counted after each treatment. The results of antimicrobial susceptibility test showed that all the tested organisms exhibited multiple antimicrobial resistance specially P. aeruginosa isolates. Survival studies showed that the tested opportunistic pathogens persisted with detectable numbers after the disappearance of the bacterial indicators which persisted maximally for 4 weeks. Treatment with UV revealed that at UVdoses 4 mJ cm2 and 8 mJ/ cm2, P. aeruginosa , A. sobria and A. caviae showed nearly similar inhibitory response. A. hydrophila was the most resistant while S. aureus was the most sensitive to UV disinfection. At UV dose 12 mJ / cm2, complete elimination of all the tested organisms occurred. The presence of multiple antimicrobial resistant opportunistic pathogens in different hospital water samples which can survive more than the bacterial indicators constitutes a potential health hazard specially for immunocompromised individuals. UV treatment has a promising effect in elimination of these organisms and it is recommended as a good method for water treatment specially for medical purposes.

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