Vol. 2, May, 2002

1/2 PERIPHERAL BLOOD-BASED PCR FOR DIAGNOSIS OF PULMONARY TUBERCULOSIS

A.M.El-Sawy, M.M.Sherief, M.F. Mohamed, M. Abdel-Hady and G.A.M. Ghanem*.

Departments of Microbiology and Chest*, Faculty of Medicine, Al-Azhar University

Tuberculosis is an important reemerging disease with increasing global morbidity and mortality. This study was performed to evaluate the detection of Mycobacterium tuberculosis DNA in peripheral blood mononuclear cells (PBMCs) using PCR technique for rapid diagnosis and follow up of treatment response in pulmonary tuberculosis patients. This work included 80 individuals who were referred to Abbassia Chest Hospital, after their consent to be included in this study during the period from August 2001 to February 2002. They were of different age groups, 52 males and 28 females, 60 of them were complaining of pulmonary symptoms suggestive of tuberculous origin (30 of them were microbiologically positive for acid-fast bacilli (positive sputum Ziehl-Neelsen "Z-N." smear and / or Lowenstein-Jensen "L-J." sputum culture) “group1”, whereas the other 30 were   microbiologically negative for acid-fast bacilli (negative sputum Z-N. Smear and L-J. Sputum culture “ group2”), the other 20 who suffered from pulmonary tuberculosis received antituberculous chemotherapy for 6 months “group3”. Pulmonary lesions were more frequent in males than females in all studied groups. Males represented 70, 60 and 65% in groups 1, 2 and 3 respectively. Pulmonary lesions showed higher prevalence in the young - and middle-age groups (“15-24” and “25-34” years) than other age groups. In group 1 Mycobacterium tuberculosis was detected in 83.3% of sputum samples by direct Z-N. Smear. Moreover, L-J. sputum culture showed higher sensitivity than direct smear as it revealed positive results in 96.7% of the same specimens. Whereas, PCR technique was the most sensitive test for detection of Mycobacterium tuberculosis DNA especially in sputum (100%), its sensitivity slightly decreased in blood samples of the same patients (90%). In group 2 all patients were negative for Mycobacterium tuberculosis by direct Z-N. smear, L-J. sputum culture, sputum PCR and blood-based PCR. They suffered from bronchitis, pneumonia, bronchiectasis and bronchogenic carcinoma. Among the 20 patients who received anti-tuberculous chemotherapy for 6 months and were negative for Mycobacterium tuberculosis using direct Z-N. smear, L-J. sputum culture and sputum PCR, 5 patients (25%) revealed Mycobacterium tuberculosis DNA in their peripheral blood mononuclear cells (PBMCs) using blood-based PCR. They were positive due to unsuccessful therapy and / or pulmonary / extrapulmonary tuberculosis. In conclusion, peripheral blood-based PCR is of value in diagnosis of pulmonary tuberculosis especially within treated subjects.

2/2 MICROBIOLOGICAL STUDY ON SOME GRAM NEGATIVE BACTERIA CAUSING URINARY TRACT INFECTIONS IN SOME EGYPTIAN HOSPITALS

M.S.E. Ashour*, A.F. Abd El-Ghaffar**, G.F.M. Gad***, M.S.M. Mancy*

and E.I. El-Hawary***

Microbiology Departments, Faculty of Pharmacy, El-Azhar University *, Faculty of Medicine, El-Minia University ** and Faculty of Pharmacy, El-Minia University***

Hospital acquired infections are among the most difficult problems confronting clinicians at hospitals. Urinary tract infections are the most frequent nosocomial infections. In this study 335 clinical strains were isolated and identified from the urine of 400 patients (83.7 %), of different ages and sexes, at El-Kasr El-Ainy, El-Sahel, Sayed Galal and El-Demerdash educational hospitals.   Of the total number of the isolates, E. coli was the most prevalent (83 strains, 24.8%) followed by Proteus species (64 strains, 19.1 %) while Hafnia was the least (6 strains, 1.8 %). The incidence of urinary infection in females was higher than in males (88.5 and 80 % respectively). The rates of urinary infection was markedly higher in elder patients than relatively young ones (97.6 and 68.4 % respectively). The present work revealed that the highest rates of urinary infection was found in patients suffering from dysuria, then those having urinary stones, and lastly in patients suffering from prostatitis (96.3, 86.9 and 57.9 % respectively). Antibiotic resistance pattern of the isolated organisms was carried out using different antimicrobial agents. Pseudomonas species were the most resistant strains against the antibiotics used. Recent quinolones including levofloxacin, ofloxacin, ciprofloxacin, and norfloxacin, showed the highest activity against all strains giving activities of 100, 98, 95, and 94% respectively. Also, aminoglycoside antibiotics as amikacin, tobramycin, and gentamicin demonstrated high activities (93, 91, and 86 % respectively), but still came after recent quinolones. Ampicillin, amoxycillin, and spiramycin showed the lowest activities (10, 14, and 20 % respectively) against the isolated bacteria.

3/2 FAMILIAL MEDITERRANIAN FEVER A SURVEY OF 80 PATIENTS, CRITERIA FOR THE DIAGNOSIS AND REVIEW OF THE LITERATURE

H.M. Moustafa, A.O. Dorrah, E. Salem* and A.M. Shandaweely**

Gatroenterology & Tropical Medicine, *Pediatric and **Surgery Departments, Faculty of Medicine, Al-Azhar University

Familial Mediterranean Fever is a genetic disorder frequently diagnosed among Arabs, Jews, Armenians and Turks. The diagnosis of Familial Mediterranean Fever still depends on the clinical ground since there is no specific laboratory test other than a molecular genetic test which is not widely available. We aimed in this study to evaluate the clinical and laboratory findings in 80 patients with FMF in Upper Egypt; to compare these findings with those in the literature, to make attention for the diagnosis and to give a brief review of the disease. 80 patients with Familial Mediterranean Fever were seen at the out-patient clinics from 1995 to 2002 and evaluated retrospectively (49 patients {61.25%} diagnosed before 1999) and prospectively (31 patients {38.75%} after 1999). A diagnostic criteria set was used in addition to the clinical and laboratory findings that can be seen in Familial Mediterranean Fever. The mean age of the study group seen at the first time was 22.53 ± 8.31 years, with male to female ratio 1.2 : 1(54.7% : 45.3%). The mean age of the patients at first attack was 6.88 ± 2.10 years for the patients below 14 years and 17.55 ± 3.21 years for the patients older than 14 years. No attacks has been occurred at the first time after 20 years. Family history was positive in 73.75% of patients. Clinical manifestations were abdominal pain (peritoneal) in 76 (95%), arthralgia and arthritis (joints) in 32 (40%), and chest pain (pleura) in 15 (18.75%) of patients. Myalgia was present in 22 (27.50%) and skin lesions in 11 (13.75%) of patients. Splenomegaly, not related to amyloidosis, was present in 13 (16.25%) of patients. Laboratory findings revealed high sedimentation rate > 40 mm/hour in 60 (75%) and leucocytosis > 10,000 mm3 in 54 (67.50%) of patients. Histopathological examination of rectal biopsy revealed amyloidosis in 6 (7.5%) of patients. Since the genetic test for identification of Familial Mediterranean Fever is not widely used in clinical application, diagnosis will remain clinical. Thus, awarness of various clinical manifestations and of the correct usage of diagnostic criteria in various patient populations is important.

4/2 PHYSIOCHEMICAL AND MICROBIOLOGICAL PROPERTIES OF IRRADIATED FORMED SURIMI DURING COLD STORAGE

S.S. Bassiouny*; F.H. Badr*; K.K.H. Ahmed**; E.A. Afifi*** and A.I. Elhanafy**

*     Food science and Technology, Faculty of Agric. Zagazig.

**   Center Laboratory for Agriculture Research Abbassa, Abou Hammad, Sharkia.

*** Nucl. Res. Center, Atomic Energy Authority, Inshas, Egypt.

The aim of the present work was to study the possibility of using gamma irradiation at dose 2 KGy for increasing the shelf life of surimi during cold storage (4+1oC) and investigate the effect of cold storage on the physicochemical, microbiological and sensory properties of irradiated formed surimi (FS). The common carp fish fillets, were minced and the obtained mince was washed and dewatered. Then mixed with cryoprotectants materials (Surimi product), the obtained surimi was filled in sheep casing formed surimi (FS) and irradiated at 2 KGy gamma rays dose. The chemical properties and the SDS-page protein fractions were determined. The obtained results showed that the chemical composition of FS and IFS were almost the same and did not alter by gamma rays. The applied dose of gamma irradiation caused a slight increase in total volatile nitrogen (TVN) and thiobarbituric acid (TBA) values. The protein fractions of FS and IFS were fractionated into 7 and 8 bands and the molecular weight of protein bands was increased by gamma irradiation treatment (2 KGy). Moreover, the total bacteria psychrophilic bacteria, yeast and mold counts were decreased after exposing formed surimi to 2 KGy gamma irradiation dose. During cold storage the microbial load of FS and IFS were raised by increasing storage periods and rejected after 2 and 6 weeks cold storage, respectively. In addition, the TVN, TBA and PH were increased during cold storage. On the other hand an opposite phenomena was observed in water holding capacity (WHC) shere force N/50 g and salt Extractable Protein (SEP) properties where decreased by increasing storage periods.

5/2 INCIDENCE OF AFLATOXINS PRODUCING FUNGI AND MYCOTOXINS CONTAMINATION IN COMMERCIAL THERAPEUTIC HERBAL MEDICINES

M.S.E. Ashour, A.A. El-Sherief and F. El-Tahan*

Microbiology Department, Faculty of Pharmacy, Al-Azhar University, Nasr City, Cairo, Egypt and * Agriculture Research Institute, Cairo, Egypt

            A total of one hundred and twenty six samples of commercial therapeutic herbal medicines were collected and examined for their fungal and mycotoxins contamination. Aspergillus flavus was isolated and examined for its capability to produce aflatoxin(s). Simultaneously, the collected samples were screened for the natural occurrence of aflatoxin(s), ochratoxin -A and zearalenone. Data showed that out of 73 strains of the isolated A.flavus, only 24 (32.8%) were capable to produce aflatoxin(s) in yeast extract liquid medium. The aflatoxin(s) producing strains were widely varied in their qualitative and quantitative capabilities to yield the four aflatoxins B1, B2, G1 and G2 at concentrations ranged between 5 to105 mg/ml. None of ochratoxin-A, aflatoxin(s) B2, G1 and G2 was detected in any of the collected samples (the detection limit was 5 mg/kg). Only aflatoxin B1 and zearalenone were Observed in 35.7 and 30% of the examined samples .The levels of concentration had a maximum of 105 and 80 mg/kg for aflatoxin B1 and zearalenone contamination, respectively.

6/2 THE ROLE OF ENDOTHELIN-1, INTELEUKIN-6 AND INTERLEUKIN-8 AS A MARKER OF THE SEVERITY OF RESPIRATORY DISTRESS SYNDROME

A. Gad*@; A. Abdel-Latif**@; A.M.E. Moustafa**@ and W. Emam ***

*: Radiological Medical Science. **: Childhood studies. @: Health Research. From Radiological Health Research Department, National Center for Radiation Research and Technology.

***: Medical Biochemistry Department, Faculty of Medicine, Zagazig University.

Objectives: The study is designed to assess the possible relationship between endothelin-1 (ET-1), interleukin-6 (IL-6) and interleukin-8 (IL-8) in tracheal aspirates and the severity of respiratory distress syndrome (RDS). Study design: We measured tracheal aspirates ET-1 concentration (TAET-1), tracheal aspirate IL-6 concentration (TAIL-6) and tracheal aspirate IL-8 concentration (TAIL-8) in preterm infants, whose lungs were mechanically ventilated; they were categorized into two groups; (RDS) group who were on mechanical ventilation for RDS and a control group, comprising prematures who were mechanically ventilated for repeated attacks of apnea of prematurity. The RDS group was further subdivided into severe, moderate and mild subgroups according to the severity of the disease based on clinical and radiological criteria. Results: TAET-1 level decreased significantly in RDS group than the control group (p<0.05). Also, a statistically significant decrease in TAET-1 was evident in both severe and moderate groups versus the control group (p<0.0001, <0.05 respectively). Statistically significant positive correlations between the mean log ET-1 and both Apgar score at 5 minutes (p<0.0001) and arteriolar/ alveolar oxygenation ratio [a/AO2] (p<0.0001) were detected. Negative correlations were found between the mean log TAET-1 and both Fraction of inspired oxygen [FiO2] and the mean airway pressure [MAP] (p< 0.0001, <0.05 respectively). Regarding TAIL-6 and TAIL-8, RDS group had higher levels than the control group (p<0.001) with both cytokines. The concentrations of TAIL-6 and TAIL-8 correlated positively with the severity of the disease. In RDS group, the mean log of TAIL-6 correlated negatively with the gestational age and a/AO2 at (p<0.0001). The TAIL-8 showed significant negative correlation with gestational age and a/AO2 (p<0.0001) and correlated positively with FiO2 (p<0.05). Conclusion: In preterm infants with RDS, lower ET-1, higher IL-6 and IL-8 in their airway effluent is pinpointing for increased severity of pulmonary status during the early course of the disease. This may be used as a predictor for subsequent complications and may help for the early treatment of preterm infants at high risk of developing complications.


7/2 RADIATION STERILIZATION OF SOME SPICES

AND MEDICINAL PLANT

H.M. Swailam and M. I. Abdullah*

Department of Microbiology and *Department of Natural Production

National Center for Radiation Research and Technology,

Nasr City, Cairo, Egypt

The effect of sterilization using 60Co gamma radiation on the microbial load, presence of some pathogens, viscosity, fatty acids and amino acids properties of some spices (caraway and anise) and medicinal plant (qarad) has been investigated. Prepackaged whole spices and medicinal plant were found to be heavily contaminated with bacteria and molds. The total bacterial counts ranged between 2.0x106- 8.2x106 cfu/g, whereas total fungal counts ranged between 4.0x103- 2.9x104 cfu/g. Qarad samples were found to be the most contaminated. All samples showed low number of the pathogenic bacterial in control. It was found that the microbiological population greatly reduced with a dose of 5 kGy and a dose of 15 kGy was required for sterilizing the samples without affecting their quality attributes. So, it can be concluded that 15 kGy of gamma radiation suffice the purpose of this work. This good marked condition was held at least during one year. Irradiation doses used linearly reduced the viscosity of all samples. These irradiation doses caused a decrease in the content of some fatty acids and amino acids and increase in others.    

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