SUMMARY
From Nanakali Hospital in Erbil city, Iraqi Kurdistan, 100 specimens were collected from aspirates, cough, and oral swabs from patients suffering from different types of cancers during chemotherapy treatments. The specimens were taken from both sexes 43 (43%) males and 57(57%) were females, the age ranged between 16 to 73 years, during the period of 10th of October to 10th of December 2022. The samples were cultured on Sabouraud’s dextrose agar (SDA) plates transferred to Mycological laboratory in the College of Science-Department of Biology-Salahaddin University. The results revealed that the number of patients who were inside Erbil city (PIE) were 59 cases (59%), while those from outside (POE) were 41 (41%). Those who had direct contact with animals (AC) were 9 cases (9%) and their ages were between 16-73 years old patients suffering from different types of cancers. The highest rate of infection was among age group (51-73) years old which was (66.6%), followed by the age group (31-50) years old with (25%), while the lowest rate of infection with Aspergillosis was at age group (16-30) years old which was (8.3%). The results showed that the highest rate of isolated fungi among infected patients was A. flavus which was 9.6%, while the infection rate with A fumigatus was 6.8%, Meanwhile, A. fumigatus was isolated only among female patients which was 5.5% (4), whereas A. flavus isolated from both sex with highest rate of infections among male patients 6.8% (5), while the female infection was 4.1% (3).
The percentage of infection with Aspergillus spp. among patients living outside Erbil city was 8 (66.6%) while 4(33.3%) reside inside Erbil city. Among 8 cases of outside Erbil city residences, 5 (62.5%) were infected with A. flavus while 3(37.5%) with A. fumigatus. Whereas 3(75%) of Erbil city residents have A. flavus and one case (25%) was infected with A. fumigatus.
The results of infected patients with Aspergillus spp. revealed that 4 (33.3%) of infected cases have been in direct contact with pet animals (chickens and cattles) while 8 (66.6%) of infected patients have not been exposed to animals at all. Regarding the percentage of cancer type and Aspergillus spp. infection cases, the results show that the highest rate of incidence was infected with different type of hematopoietic malignancies, 5(41.6%) with Acute lymphoblastic leukemia (ALL), 2 (16.6%) chronic lymphocytic leukemia (CLL), 2 (16.6%) multiple myeloma (MM), and 1 (8.3%) lymphoma. Others were infected with 1 (8.3%) case of colon cancer and 1 (8.3%) case of lung cancer. On the other hand 3 cases of A. flavus infected patients have ALL, 2 cases have CLL, while only one case for each patient with MM, colon cancer and lung cancer were observed. On the other hand two cases with A. fumigatus infection have ALL, one has MM, and the last case has Lymphoma.
The highest rate of fungi isolated was A. flavus 9 (45%), followed by A. fumigatus 5 (25%), Penicillium spp. 4(20%) and lowest rate was Mycelia sterilia 2(10%). The identified Aspergillus spp. by using three selective media were: 8(40%) were A. flavus, 4(20%) were A. fumigatus.
The identified A. flavus and A. fumigatus were subjected to molecular identification conformation depending on PCR technique using universal primers gene amplification, Sequencing examination of several amplification products revealed (99-100%) sequence identity with the Aspergillus strains. Sequence data of isolates provided GenBank accession number, for nucleotide sequence.
The phylogenetic tree revealed that A. flavus and A. fumigatus are closely linked to other species of Aspergillus. Based on the alignment of current and global reference sequences, our A. flavus OQ380845, and A. fumigatus OQ380844, were closely related to each other.
Sensitivity test by disc diffusion method and MIC microtiter plate method were applied for the isolated Aspergillus to detect the inhibitory effect of the antifungal drugs that used in Erbil city’s Hospitals against A. flavus and A. fumigatus.
The ANOVA test showed significant differences in the sensitivity of all tested species against selected antifungal discs. In general A. flavus and A. fumigatus were sensitive to Econazole, moderately sensitive to Ketoconazole and miconazole while less sensitive to Nystatin, To detect MICs of antifungal drugs: Fluconazole, Terbinafine and Nystatin, with Concentration (500mg/ml), (150mg/ml), (250mg/ml), and (500,000 IU) respectively on A. flavus and A. fumigatus, the 96-well a flat bottom microtiter plate method were performed. After 72 hours of 27°C incubation, the OD value of each well was measured at 490 nm using an Enzyme-Linked Immunosorbent Assay (ELISA) reader, Fluconazole at (2.35 mg) show the moderate sensitivity to A. flavus and A. fumigatus Our study show Econazole is more effective antifungal drug on A. flavus and A. fumigatus than other antifungal drugs.