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
Antibiotic resistance is a problem of deep scientific concern both in hospital and community settings. Rapid detection in clinical laboratories is essential for the judicious recognition of antimicrobial resistant organisms. So, the growth of Uropathgenic Escherichia coli (UPEC) isolates with Multidrug-resistant (MDR) and Extensively Drug-resistant (XDR) profiles that thwart therapy for (UTIs) has been detected and has straight squeezed costs and extended hospital stays. This study aims to detect MDR- and XDR-UPEC isolates. Out of 42 UPEC clinical isolates were composed from UTI patients. The bacterial strains were recognized by standard laboratory protocols. Susceptibility to antibiotic was measured by the standard disk diffusi
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This work deals with the description of histopathological effects of the nematode Hartertia
gallinarurn Theiler. 1919 on the digestive system of the seesee partridge collected from Qa’ra
area in the western desert district of Iraq. along with some notes on intensity fluctuation of the
parasite according to the seasons. It is found that the major effects of the nematode are
necrosis and fibrosis of gizzard: granulomatous reaction. necrosis and mononuclear
infiltration of proventriculus: damage of mucosal lining of intestine and lymphocytic
infiltration of liver.
A pulsed (TEA-0O2) laser was used to dissociate molecules of silane ethylene (C2I-14) and ammonia (NH3) gases, through collision assisted multiple photon dissociation (MPD) to deposit(SiC i_xNx) thin films, where the X-values are 0, 0.13 and 0.33, on glass substrate at T,----648 K. deposition rate of (0.416-0.833) nm/pulse and thickness of (500-1000)nm .Fourier transform infrared spectrometry (FT-IR) was used to study the nature of the chemical bonds that exist in the films. Results revealed that these films contain complex networks of the atomic (Si, C, and N), other a quantity of atomic hydrogen and chemical bonds such as (Si-N, C-N, C-14 and N-H).Absorbance and Transmittance spectra in the wavelength range (400-1100) nm were used to stud
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The detection of fungi contaminating maize grain and the effect of four plant extracts Azadirachta indica, Eucalyptus globulus Glycyrrhiza glabra and Zingiber officinale on the growth of A. flavus and its ability to produce AflatoxinB1. The results showed that the incidence of Aspergillus spp., was 52.75% of the isolated fungi, of which 29.50% was due to Aspergillus flavus, followed by Penicillium spp., with an incidence of 21.06%, and then Fusarium spp., with a rate of 18.13%. The percentage of toxin-producing A. flavus isolates reached 70.8% out of 24 isolates. The results showed the effect of alcoholic plant extracts at a concentration of 10 mg/ml on the fungal growth activity of A. flavus, the alcoholic extract of neem leaves was superi
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The objective of this study was to progress another method for coagulation/flocculation of the microalga Chlorella vulgaris via pellet-forming of the fungal species Aspergillus niger which was isolated from municipal wastewater mud and the facultative heterotrophic microalga "C.vulgaris was used. The main factors studies were spore inoculums, organic carbon concentration in medium as well as pH variation which had considerably positive effects on microalgae/fungi co-pelletization formation. The process parameters are an inoculum1×104 spores/ML, 15 g/l sucrose as carbon source and pH ranged from 5 - 7.0 were found optimal for efficient microalgae/fungi co-pelletization formation. For autotrophic growth, when pH of culture broth was adjusted
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In the leaves of Olea europaea L. Olive trees an endophytic fungus was discovered. Cladosporium sp. was identified to be the fungus based on its morphological characteristics and nuclear ribosomal DNA ITS sequence analysis and was registered in NCBI as the Cladosporium genus has been registered under the number (0P939922.1) The species was not specified, and it was considered of unknown species after comparing it to global isolates. In comparison to olive leaf extract, Cladosporium sp. including total flavonoid, total phenolic, total terpenoid, and total saponins, Which were 121.9%, 198.1%, 89.13%, and 29.87 % respectively compared to its content in olive leaf extract, which was 61.54 %, 67.88 % , 17.1
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The study is concern on determine the type of Candida spp.in leukemia patients that were infected with oral candidiasis as a result to their immune suppression (weekend immune system) due to their submission to radiation and chemotherapy treatment. The result showed that the most common isolates were C. guillermondii 19 which represent 31.66% of cases, then followed by C. itermedia 11 which represent 18.3%, while the less common isolates were for C. zeylamodes 3 which represent 5%.
The predisposing factors and their effect on the increasing of vulvovaginal Candidiasis have been studied in this research. The result indicated that the highest of infection with vaginal Candidias was in the age of women with age of (21-30years) with vaginal itching and discharge, also the results revealed that the pregnancy was the main predisposing factor. The percentage of the infected pregnant Ladies was 33.7% followed by the women who used the IUCD and OCP 22.1%, those who used steroids 5.2% comparing with control women (without predisposing factors).The results also indicated that the most cases of infection were during the third trimester of pregnancy (where the rate of infedion) was 53.9%.-OCP- Oral Contraceptive Pills-IUCD- Intrr
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