Abstract Candida albicans is a commensal fungal pathogen that grows in yeast and hyphal forms in the human gut. C. albicans causes mucosal and cutaneous diseases that can result in significant mortality following systematic infections and it also exhibits drug resistance. Zebrafish have been an excellent model to investigate C. albicans infections because of their transparency and the availability of many transgenic lines. However, there is a limitation in using zebrafish as a model because the fish embryos cannot survive at 37°C therefore it is not suitable for studying Candida infections at physiological relevant human body temperature. In this thesis, the normal embryonic development of Arabian killifish (A. dispar) is investigated, revealing that embryogenesis was divided into 32 stages based on diagnostic patterns of development. A. dispar can also found to tolerate a wide range of temperatures and salinities. This suggests that A. dispar could be developed as a novel model to investigate host-pathogen interactions. The tolerance of A. dispar to high temperatures may in part be attributable to brown pigment cells with a highly fluorescent character that may have developed to allow the fish to adapt to live within extreme environmental conditions with strong sunlight and a wide range of temperatures (Chapter 3). In terms of Candida infections, this study examined A. dispar as a model to test C. albicans pathogenicity. The survival of A. dispar embryos following Candida infection showed a dose dependent relationship. We also found that A. dispar can survive longer than zebrafish after infection. Furthermore, C. albicans cells were observed to undergo a transition from yeast to hyphae at 37°C. An investigation of the ability of mutant strains of C. albicans with defects in cell wall mannosylation revealed a significant impact on virulence, host mortality, and the fishes’ immune response. The present study found that although the deletion of O- and N-mannan from the cell wall of C. albicans, affected fungal burden (attenuation), and the survival of the infected embryos per se was significantly decreased in the infections of the mutant strains compared to the WT. This data confirms the importance of the mannosylation state of the cell wall in triggering an immune recognition event (Chapter 4). A. dispar is also shown to be suitable for studying the effectiveness of 3 | P a g e antifungals. Fluconazole treatment of infected embryos and eggs promoted greater rates of survival at high doses, alongside a significant reduction of C. albicans CFUs (Chapter 4). When looking at the Candida-host interaction, we directly observed phagocytosed yeast cells within macrophages. Various detection methods were used to follow macrophages and neutrophils including Western blotting, immunostaining and histological staining (Sudan black and FITC-tyramide) allowing the monitoring of the time course of the immune cells. A biphasic response of macrophages was detected by L-plastin Western blotting, suggesting activation of two different type of macrophage: activated macrophage (M1) and alternative macrophage (M2). We also assayed reactive oxygen species (ROS) within infected embryos using a fluorescent probe (H2DCFDA), revealing the accumulation of the fluorescent probe at the sites of infection. Quantitative and qualitative analyses of the oxidative and immune response using the H2DCFDA and qPCR were also accomplished within A. dispar embryos after infection with both the WT and mutant strains of Candida albicans (WT, pmr1∆, mnt1-mnt2∆, and och1∆). The results confirmed that the mutant strains did not activate a host oxidative stress response nor immune cell accumulation when compared to WT, suggesting that the immune response is less activated against these mutants. Finally, a new transgenic line of A. dispar fish was developed using Betaactin-DsR-LoxP-GFP. The new transgenic A. dispar is suggested to be an ideal model for real time observation of host-pathogen interactions and for investigation of molecular functions of the immune response. Overall these results improve our understanding of the use of a new transparent fish model to study fungal pathogenesis and demonstrates the potential advantages of using this species in future studies of bacterial, fungal and viral pathogens at a physiologically relevant temperature for human infection. Such a model could lead us to investigate in more depth the key interactions between pathogens and their host and permit the screening and development of new antifungal therapies (that might target the pathogens directly or target the host immune system). View full metadata
This work aimed to investigate the prevalence of pathogenic fungi and evaluate the antifungal activity of Trichoderma orientale FMR12486 crude extract against pathogenic fungi isolated from patients attending the National Center for Thoracic and Respiratory Diseases (having a history of tuberculosis) and consultant of Dermatology of Baghdad hospital, Iraq. A total of 80 clinical specimens were collected: 20 skin scrapings specimens and 60 sputum specimens. The results of direct examination by KOH 10% and culture showed that 11 (55%) cases from 20 skin specimens were positive for fungal infections, while in the sputum specimens, 28 (47%) cases from 60 were positive. Candida albicans represented the most common fungal infection isolat
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In addition to being a religious book with high human and moral themes, Nahj al-Balagha is considered a mirror of Arab culture and a literary masterpiece at the height of eloquence and eloquence, and because proverbs in the form of short, concrete and understandable phrases for everyone, experiences, thoughts and convey ideas, Imam Ali (AS) used it to facilitate the understanding of various political, social and moral concepts. In this article, we intend to criticize the way Dashti, Shahidi and Foladvand translated it by using Newmark's model due to the importance and cultural reflections of proverbs in understanding the cultural atmosphere governing Nahj al-Balagheh. In his evaluation model, Newmark divides cult
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The present study was designed to investigate the effect of R. stolonifer metabolic products on some antioxidant defenses and some elements (Cu, Fe, Ca) . The experiment was performed using 25 mature male rats, their age average was about (3-3.5) months and their weight average was about (200-225) gm. The animals were randomly divided into 5 equal groups (five animals for each): control (treated orally with normal saline) and four treatment groups were drenched orally with four concentrations of R. stolonifer metabolic products (15, 30, 60,120) μl/kg body weight. The animals were treated with one single dose of the previously described concentrations then left for 15 days. Animals of different groups were sacrificed under light
... Show MoreM. domestica is the most important insect that transmit pathogens for diseases in the world. The use of nanotechnology is eco-friendly method in control pests. The study aims to investigate the feasibility of bio-manufacturing nanocapsules of fungal secondary metabolites in order to improve the efficiency of metabolite and assess their inhibitory effect on the acetylcholine esterase enzyme in housefly larvae. An equal mixture of organic solvents, ethyl acetate and dichloromethane, was used to extract the metabolic products of the fungus M. anisopliae, (PEG4000) and chitosan was used in the preparation of nanocapsules. The results of the DLS granular size assay showed that the size of the extract particles and the size of the chitosan and
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Microbial antibiotics resistance is considered a serious health issue in the Middle East and developing countries. In this study, the Fe2O3 nanoparticles was prepared chemically, and the particles size and shape were analyzed by using Scan electron microscope (SEM) and X-Ray diffraction (XRD). Different concentration of Fe2O3 nanoparticles were used and examined on E.coli and S. aureus. Using liquid dilution and in vitro cytotoxicity assay by microplate toxicity test (MTT). The microbial cell metabolic activity was measured on gram-negative, gram-positive bacteria and fungi after treating with different concentrations of Fe2O3 nanoparticl
... Show MoreOpportunistic fungal infections due to the immune- compromised status of renal transplant patients are related to high rates of morbidity and mortality regardless of their minor incidence. Delayed in identification of invasive fungal infections (IFIs), will lead to delayed treatment and results in high mortality in those populations. The study aimed to assess the frequency of invasive fungal infection in kidney transplant recipients by conventional and molecular methods. This study included 100 kidney transplant recipients (KTR) (75 males, and 25 females), collected from the Centre of Kidney Diseases and Transplantation in the Medical City of Baghdad. Blood samples were collected during the period from June 2018 to April 2019. Twent
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Background: Urinary tract infections (UTIs) and their complications such as Bladder cancer (Bl. C.) are a health growing problem worldwide. Objective: To shed light on this subject, present study was done to investigate relationship between recurrent urinary tract infection (RUTI) due to Escherichia coli (E. coli) and Bl. C.Type of study: Cross-sectional study. Methods: This study included 130 patients with RUTI, 50 patients with Bl. C. and 50 control of both sexes (aged 7-85 years) attending Al-Zahra Teaching Hospital in Al-Kut/Wassit governorate and Al-Harery Teaching Hospital of specialized surgeries/Baghdad. The patients were divided into two groups: the first group (n=130) included those who were suffering from recurrent UTI without
... Show MoreThe aims of this study were to prepare Cryptococcus neoformans killed whole cell antigen and evaluate their effects by using DTH- skin test and the differential white blood cells in local rabbits.
Fourteen animals of both sexes were divided randomly into two groups. The first group (7 animals) was immunized with 1 ml of Cryptococcus neoformans killed whole cell antigen (1x 10 cells/ml), subcutaneously. A booster dose was given after 14 days of the first dose. The second group (7 animals) was considered as control group. Based on results of DTH-skin test, no significant differences (P< 0.05) were recorded between the concentrations, 15 mg/ml and 7.5 mg/ml after 24, 48 and 72 hrs., but there was a significant differences (P<0.05)
Background: Morganella morganii is one of the important nosocomial pathogens that may cause urinary tract infection and bacteremia.Methods: The above bacterium was identified from 250 bacterial strains which were isolated from 220 urine samples of patients with urinary tract infection. Antimicrobial susceptibility, by using disk diffusion method, of isolates was tested against some antibiotics.Results: Two M. moganii strains were isolated from female catheterized urinary tract patients, and identified by conventional biochemical tests and API20E system at the first time in Iraq. Both of them produced urease and hemolysin. Antimicrobial susceptibility test showed that these strains are resistant to, amoxicillin-clavulanate, cephalothin, g
... Show MoreThis in vivo study was conducted to investigate the effect of different concentrations of ozonated water on experimentally skin infection with some of bacterial isolates (Pseudomonas aeruginosa and Staphylococcus aureus ) which Showed dermal infection in experimental animals after 48 hours of exposure to these Microorganisms. Results revealed that ozone has the power to accelerated the healing process depending on the perfect concentration of ozonated water used and the severity of infection & nature of causative agent , in which the recovering period was 7 days for the infection caused by P. aeruginosa and 5 day for S.aureus by using the concentration 60 µg/ml . Results also indicated in this study the stimulated effect of O
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