PHD theses : Medical Laboratory Science
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Item GENOME WIDE SEARCH FOR VISCERALLEISHMANIASIS SUSCEPTIBILITY GENESIN SUDANESE POPULATION(ALNEELAIN UNIVERSITY, 2005-10) MANAL AHMED MOHAMED AHMED FADLABSTRACT Sudan is one of the major foci of visceral leishinaniasis (VL) globally (the other two foci are Brazil and India). Familial clustering and ethnic differences suggest genetic factors may be involved in the infection. In this study a two stages genome-wide scan was employed using two independent sets of families from two villages: El-Rugab and Um—Salala. located 40 kilometers apart in the heart of the endemic area of eastem Sudan. These villages are inhabited by the Masalit ethnic group who migrated from westem Sudan in the 1980s. In the first stage (= scanl) we genotyped 400 highly polymorphic microsatellite markers (approximately 10 cM apart) in 220 individuals from 38 multicase pedigrees (= scanl families) comprising 48 nuclear families (1 18 affected sibs). Scan] was followed by grid tightening strategy in which 35 additional markers were added close to regions that gave suggestive evidence for linkage of VL susceptibility in scan 1. These markers were typed in scanl families. ln the second stage (scan2). 20 markers from positive regions of linkage and the 35 additional markers were all typed in family set 2 (=scan2 families: 21 nuclear families, 48 affected sibs). ln scan 1, the multipoint analysis performed provided evidence for linkage of VL susceptibility to 5 regions on chromosome lp22, ]q3l.3. 5q34-35.3, 6q27 and l3q 31.1 (0.0002Item Laboratory Diagnosis of Male Infertility among Sudanese Couples(Alneelain University, 2006) Abdalmula Mohammed Abdallaln view of the increasing number of infertility among the couple male present in the Sudan .This study was conducted to essentially study the factors responsible for decreased male fertility, the impact of toxic and antioxidant elements on male reproductive function .and to introduce biochemical markers to be used in male infertility diagnosis . Group of 500 infertile male subjects were enrolled ;15O azoospermic 150 oligospermic patients, 100 patients with abnormal sperm motility and 100 patients with abnormal sperm morphology were studied, and compared with reference group of 100 fertile male. The fertility and thyroid hormones were measured by ELISA and trace elements in semen and serum/blood by AAS. Semen volume and liquefaction time in azoospermic and oligospermic patients was significantly reduced (p< .05 ). Significant correlation was noticed between both FSH & LH with seminal volume , and blood Pb with liquefactions time in azoospermic patients .Significant correlation was observed between seminal Zn and liquefaction time in both azoospermic and oligospermic subjects.(r < .05). Immunological infertility is higher in patient with abnormal sperm motility (9% lgC-3, 6% lgA) than other test groups. Testosterone in the test groups was significantly reduced , while FSH level was significantly increased (p<.O5). Prolactin level was increased in azoospermia, and significantly increased in the other infertile groups (p<.O5). LH was significantly increased in both azoospermic and oligospermic patients (p<.O5) Significant correlation was noticed between ,LH and semen Cd in azoospermia , prolactin was significantly correlated with Pb in both blood and semen, and with seminal Mn and Zn, LH with seminal Zn in oligospermic patients. Also correlation between, prolactin with blood Pb and FSH with blood Cd in abnormal sperm motility patients (r<.05), .The thyroid hormones showed conflicting levels in the infertile test groups, only T4 was slightly increased. In seminal plasma Cd level was significantly increased in both azoospermic and oligospermic patients (p < .05). Seminal Pb level was slightly elevated in infertile patients with azoospem1ia and oligospermia . Zn level was significantly reduced in the infertile groups (p<.O5). Se , Mg & Ca were significantly reduced in both azoospermic and oligospermic patients (p<.O5). Where as Mn, Ni, Co,Cr and Cu showed no significant variations .ln blood Pb level was increased in the test groups, and Cd level was significantly increased in azoospermia (p<.O5), and the other elements indicated conflicting level , _but within normal range when compared with control group. Pb level was significantly correlated in blood and semen of azoospermic and oligospermic subjects,Also Cd level was significantly correlated in both semen and blood of abnormal sperm motility patients (r<.O5). Seminal NAG was significantly reduced in the study groups , while citric acid was reduced in both azoospermic and oligospermic patients (p<.05).Semen fructose indicated conflicting level ,but within normal range .Significant correlation was noticed between NAG with both FSH and LH in azoospermia, and with prolactin and seminal Mg in oligospermia (r<.O5).2.7% of azoospermia had sertoli-syndrome , explained significant decreased in testosterone ,seminal NAG, citric acid Zn, and increased FSH. (P<.O5). 13% of the infertile have varcocele, recorded significant increased FSH,LH and decreased seminal NAG ,citric acid 8- Zn (p<. 0 5). 9.6% of the test groups had erecting and ejaculating problems. Characterized by significant increased in prolactin and Mn level.7.2% of the test groups are smokers, explained significant increase of both Cd and Pb in blood and semen, and reduced semen volume, NAG and Zn level (p<.05) Treatment of adult male rats with Cd, Pb caused hair loss and enlargement of the testes. Followed by significant increased of blood Cd , blood Pb, FSH and LH (p<.05), decreased testosterone and Zn (p< .O5).With blockage of spermatogenesis at seminiferous tubules level, maturation arrest ,and proliferation of the sertoli cell. Where as treatment with Zn, showed increased number of germs cells and developing spermatide .Mixing of Cd with Pb in one dose exacerbated the toxic action of this elements, caused decreased testosterone and Zn, increased FSH , LH and prolactin with deletion of the germ cell, and proliferation of the sertoli cells . While mixing of Zn with Cd and Pb , reduced the toxicity of this elements on fertility hormones, development of the germ cell, and proliferation of sertoli cellsItem Molecular Genotyping of Methicillin Resistant Staphylococcus aureus in Khartoum Teaching Hospital(Al Neelain University, 2007) Eidha Ali Awadh Bin HameedThis is a cross—sectional study perfomied to detect the methicillin resistant Staphylococcus aureus (MRSA) isolates collected from Khartoum Teaching Hospital in the period from September 2005 to August 2007 by various molecular typing methods. A total of 48 S. aureus isolates were collected fiom clinical wound specimens in wards of surgery, orthopaedic, and burns. The number of MRSA was found to be 9 (l8.75%). The isolates were classified into 3 groups; group 1: S. aureus isolated from the surgical ward (28; 58.3%), group H: S. aureus isolated from the orthopaedic ward (14; 29.2%), and group IH: S. aureus isolated from the bums unit (6; 12.5%). All strains were studied for their susceptibility to commonly used antibiotics. The results revealed that the drug of choice for the treatment of nosocomial MRSA and MSSA is vancomycin. While multi-drug resistance was observed to be common among MRSA strains. Polymerase chain reaction (PCR) was used to amplify the sequence of S. aureus specific gene at (107 bp) all isolates. Methicillirt resistant (mec/1) gene yielded the amplicon for size at 1319 bp of 9 isolates, and coagulase (coa) gene produced amplification products approximately at 500 bp (26/48), and sso bp (22/48). Two distinct PCR-restriction fragment length polymorphism (RFLP) pattems of coagulase gene exhibited among isolates of S. aureus; coaA and coaB. With Alul restriction enzyme digested, the product fragments were approximately at 190, 310 bp and 190, 390 bp with percentages of 54.2% (26/48) and 45.8% (22/48) respectively.Item Isolation, Identification and Molecular Characterization of Salmonella Species From different sources(Al Neelain University, 2009-02) Abbas Hassan MohammedA total number of 733 samples were collected fiom different sources (clinical, food, poultry and water), and subjected to bacteriological examination for isolation of Salmonella spp. using enrichment and selective media. The percentages of the detection of the organism from the above sources were 10.5, 2.9,‘ 2.5 and 0.0 respectively. y From the 200 feacal specimens (100 from human and 100 from animals),'the organism was detected in 20% of the human specimens and only as 1% from the animal specimens. Food specimens revealed isolation of Salmonella spp as 10% and 1.85% from meat and milk specimens respectively, while percentages of 5.1 and 1.6 were obtained from chicken and eggs specimens and the raw shaworma revealed negative growth of the Salmonella spp. i Using the API method for confinnation of the isolates gave unacceptable profiles with 66.7%, 44.4% and 66.7% for Salmonella spp isolated from clinical, food and poultry specimens respectively, (in all unacceptable profile organisms the Salmonella spp was the first suggested organism). 41.7% of the total specimens showed confirmation of the presence of the organism with identity of 99.9%. The API also showed that there are strong similarities in biochemical reactions of the Salmonella spp K. pneum. Ozaenae, E. coli 2, E. coli 1 and Hafnia alvei. The serological investigation revealed the identification of Salmonella anatum from both poultry and food specimens, Salmonella allerton from both poulhy and human specimens and Salmonella enteritidis from both food and human specimens. Salmonella Kentucky, Salmonella albertslunal Salmonella abortus bovis, Salmonella tchad, Salmonella II. Other serotypes were detected in human specimens without detection for the common known pathogenic species of Salmonella typhi and Salmonella paratyphi. Food specimens also revealed the presence of Salmonella okerara, Salmonella Harrisonburg and Salmonella maiduguri. _ l All isolates fiom the different sources showed high sensitivity towards Arnykacin, Ciprofloxacine, Cefoperazone and Netilmicin antibiotics.‘ Most of the isolated Salmonella spp showed clear resistance toward Augmentin antibiotic. There were clear variations in the susceptibility of Salmonella spp toward different antibiotics according to their origin of source. The PCR product analysis indicated the presence of the gene associated with virulence in two of the two Salmonella enteritidis serovars with plasmid profiles containing molecular weights of 26, 424 and 51, 418 kb. The Salmonella enteritidis also showed the presence of virulence gene as single amplification product of 460 bp.Item Isolation and ldentificationof Clostridium difficile and Other Clostridium Species Associated with Diarrheic Children and Their Susceptibility to Antimicrobial Agents(Al Neelain University, 2007-09) Susan Abdul-lateefShareefClostridium species is part of the intestinal indigenous microbiota of young children and they can produce several endogenous infections Clostridium difiicile frequently colonizes the human large intestine when the normal colonic flora is disturbed by antibiotic therapy. The result of colonization may be asymptomatic, or it‘ may lead to illness, ranging from mild diarrhea to pseudo-membranous colitis. C. dlflicile is a major nosocomial pathogen, responsible for up to 20% of cases of antibiotic- associated diarrhea in industrialized countries, and is an emerging problem in developing countries. The study concerned 552 children, aged 15 days and 8 years. 351 fecal samples were taken from diarrheic children, and (201) were taken from non- diarrheic patient children carrier of C. difiicile who were at risk in whom CDAD may develop after using chemotherapeutic agents. Sixty-two isolates of Clostridium spp. were characterized by colony morphology on several media, spore shape and position, biochemical tests and GLC technique. They identified as: Clostridium septicum 5 (8.1 %), C. bifermentans ll (17.7 %), C. s0rdellii4 (6.45 %), C. perfringens 5 (8. 1 %), C. novyi 8 (13 %), C. botulinum 10 (16 %), C. sporogenes 13 (21 %), C. tetani 3 (4.8 %) and C. tertium 2 (3.2 %). One strain of Clostridium diflicile (1.61 %) was detected on selective medium (CCFA). A total of 202 cooked meat carbohydrates selective (CMC+S) broth indcillated with fecal samples, were tested for detection of Clostridium I514 ll 1 VIII .,l it-l 14¢ ll 1- 1» Hr» I I diflicile using C. diflicile antigen detector (latex agglutination test); 14 (7 %) of them were positive. Fatty acid analysis by gas-liquid chromatography technique was used for detection of isocaproic (iC6) peak in height of Z 2mm, and also valeric and isovaleric acids. We found that 13 of them (93%) yielded height peaks of iC6 acid Z 2 mm, while one of them yielded height peak < 2mm. The isolates were tested against eight kinds of antimicrobial agents included: Metronidazole, Tetracycline, Vancomycin, Naldilic Acid, Trimethoprim, Erythromycin, Kanamycin, Gentamycin and Clindamycine. The inhibition zone diameter showed that the most effective antimicrobial agents was metronidazole, followed by erythromycin and Naldilic acid, while most of species revealed no inhibitory zone against trimethoprim. There were high levels of multi-drug resistance among the clostridia isolated. This may be attributed to the irrational use of these chemotherapeutic agents.Item A New Simple And Cheap Method For The Eradication Of Transfusion — Induced Malaria In Sudan(Neelain University, 2004) Mohamed Siddig Mohamed AliABSTRACT Background: Transmission of malaria through blood transfusion is a genuine problem for the capability of plasmodia to survive in stored blood and the weakness of transfused patients. Systemic screening of blood donors is not a practical solution of the problem even by using an advanced technique as well as treatment of transfused patients after transfusion and/or prospective donors before donation. Therefore, an ideal way for such prevention could be to kill the parasite in donors’ blood before transfusion. Objective: To select the antimalarial drug of choice that can be routinely added to donors’ blood in vitro to kill malaria parasites within maximum three days. Material and Methods: Donors’ blood which was collected from 4484 blood donors has been screened for malaria parasite microscopically using Giemsa’ staining technique. Of these samples, only 30 (500ml of blood each) satisfied the inclusion criteria of this study. Each of these blood samples was subdivided equally into ten sub-samples to obtain a total of 300 sub- specimens. Three concentrations of each antimalarial drug (chloroquine, fansidar and quinine) were added to 30 specimens while 30 specimens (control) were lefi without adding antimalarial drug. Blood specimens were tested for parasite culture, platelets count, total leucocyte count, packed cell volume, lysis percentage, osmotic fragility, prothrombin time, activated partial thromboplastin time, sodium and potassium serum levels simultaneously on the day of collection. Thereafter, it was stored in the blood bank refrigerator (4°-6°C) and tested after 24 & 48 hours by the same laboratory procedures. Results: The reduction of malaria parasites numbers was found to be proportional to the concentrations of chloroquine, fansidar and quinine added to donors’ blood and to the storage period. Whereas, the control donors’ blood samples (without antimalarials) revealed stable number of the parasites even after 48 hours storage. Fansidar was highly effective afler 24 hours storage followed by quinine and 48 hours storage revealed high effectiveness of fansidar followed by quinine and chloroquine. The optimal doses of the applied antimalarial drugs were generally safe to all constituents of the stored blood compared to the control blood samples. Conclusion and Recommendation: It was concluded that for eradication of transfusion induced malaria by in vitro processing of donors blood, fansidar is the best drug that can be used followed by quinine. So it was recommended to apply the optimal doses of these drugs to the components of the blood bags before phlebotomy.Item SERO- DETECTION OF HEPATITICS B, HEPATITICS C AND SALMONELLA MICROORGANSMS AMONG SUDANESE SCHISTOMIAASIS PATEIEN IN GEZIARA(Neelain University, 2006) MAZIN OMAR MOHAJERThis cross-sectional study was conducted to determine the relationship between schistosomiasis , salmonellosis, Hepatitis B, and Hepatitis C infections in Helate Mahajoub in Gezira Agricultural Scheme . ' Idrine and faecal samples were collected from 288 individuals and‘ inpyestigatcd for Schistosomaova using the modified Kato thick smear technique and the concentration technique. A serosurvey of 207 subjects was conducted to diagnose S.typlzi, S.paratypI1i A, and S.paratyplzi B using the Widal test technique. Hepatitis B surface antigen (HBsAg) and Hepatitis C antibodies were estimated using the immuno chromato- graphic technique (ICT). The scro-positivity of HBsAg and HCV antibodies were 21.7% and 9.1% respectively; while enteric fever showed a 30.9% sero- positivity. i The association established in this study may be explained by the immune status of the infected subjects studiedItem Isolation and Identification of Clostridium difficile and Other Clostridium Species Associated with Diarrheic Children and Their Susceptibility to Antimicrobial Agents(Neelain University, 2007) Susan Abdul-laIeef ShareefClostridiznn species is part of the intestinal indigenous microbiota of young children and they can produce several endogenous infections Closlridimn difficile frequently colonizes the "human large intestine when the normal colonic flora is disturbed by antibiotic therapy. The result of colonization may be asymptomatic, or it may lead to illness, ranging from mild diarrhea to pseudo-membranous colitis. C. difliqile is a major nosocomial pathogen, responsible for up to 20% of cases of antibiotic- associated diarrhea in industrialized countries, and is an emerging problem in developing countries. The study concerned 552 children, aged l5 days and 8 years. 351 fecal samples were taken from diarrheic children, and (201) were taken from non- diarrheic patient children carrier of C. difiicile who were at risk in whom CDAD may develop after using chemotherapeutic agents. Sixty-two isolates of Clostridizun spp. were characterized by colony morphology on several media, spore shape and position, biochemical tests and GLC technique. They identified as: Closrridium septicum 5 (8.1 %), C. bifermenlans 1 1 (17.7 %), C. sordell/‘i4 (6.45 %), C. pe/fri/1gens5 (8. l %), C. novyi 8 (13 %), C. botulimmz 10 (16 %), C. sporogenes l3 (21 %), C. telani3 (4.8 %) and C. rerliu/n 2 (3.2 %). One strain of C10slria'izun difficile (1.61 %) was detected on selective medium (CCFA). A total of 202 cooked meat carbohydrates selective (CMC+S) broth inoculated with fecal samples, were tested for detection of Closlridium