Isolation and Identification of the Fungi that Causes Eye Infection in Wasit Province, Iraq
Noor Reyadh Jameel1, Abdul Basit Abdul Samad Abdulla2, Hussain Ali Abd3
Department of Microbiology, Collage of Medicine, University of Wasit /Iraq [email protected]
ABSTRACT
Fungal ocular infection is one of the most important infections in Iraq inspite of it occcure in low incidence in compare with bacterial and viral eye infections. Two hundred(200) eye swabs samples were taken from patients suffering of eye infections.
After culturing on the related selective media, results showed that 29 (15%) of the samples were positive for occurrence of microorganisms; of these, 26 (91%) were belonged to mold and 3 (9%) yeast. After the fungal isolates were identified by cultural and microscopic examinations, the following species and percentages were recorded: Aspergillusflavus 13 (45%), Aspergillusterrus 5 (17%), Aspergillusfumigatus4 (14%), Aspergillus niger 2 (7%), Penicillium spp. 1 (4%) and Alternaria spp.
1 (4%). While the yeast isolates were belonged to Candidaguilliermondii 1(3%), Candidaciferrii 1(3%) and Cryptococcuslaurentii 1(3%) according to their identification through the cultural, microscopic examinations, in addition to the biochemical examinations by using VITEC 2 system. Upon such findings, Aspergillusflavus was the predominant fungi.
Regarding to the patients gender, it was found that cases of fungal eye infections in males two fold than in females. Patients group of age (50-59) years was the most infected group. Tobacco smoking and Diabetes have positive effects on infections, while there was no effects for blood pressure.
Keywords
Aspergillus flavus, Aspergillus terrus, Aspergillus fumigatus, Aspergillus niger, Penicillium spp. ,Alternaria spp., guilliermondii, Candidaciferrii,Cryptococcuslaurentii, VITEC 2 system.
Introduction
The eye is the most exposed part in the body to the air thatcarried dust and microorganisms such as bacteria and fungi that causing infections to it, specifically when any scratch or wound tack place in the lining tissues of the eye, as these microorganisms immidatly Settle down into the tissues that broken, resulting prominent damage to the eye(Hameed, 2020).Infections with fungi is a significant ophthalmic issue in tropical and subtropical region, although, it occasionally occur in other high-income countries with temperate climates.The most common fungal pathogens are Aspergillus spp. ,Fusarium spp. and dematiaceous spp.(Garget al., 2016 ; Soleimaniet al., 2020). The factors that are risk for getting fungal infections are: ocular trauma, ocular surface disease, uses of contact lens, uses of topical steroid and systemic immunosuppression (Erdemet al., 2017 ; Kumaret al., 2019). Treatment of infection has become a problematic case, particularly in sever problem, because of a few available antifungals and emerging of resistance species. On the other hand, a failure to treat can result in the vision loss (Soleimaniet al., 2020). This study was aimed to isolate and identify the fungi that causative of eye infection in Wasit province .
Literature Review
The human eye is one of the most noticeable sensory systems.The majority of the information about the external environment gathered byHuman beingsthrough their eyes and thus depend on eyesight more than other sense, with the eye being the most sensitive organ we have (Lee ,2020).
Eye Infection
Thecommensal organisms can be seenon the external ocular surface, such as Staphylococcus species, Corynebacterium species, and Propionibacterium species, which are form the resident flora. Any environmental organism can be a transient flora in the eye by achance. The intraocular tissues and spaces, however, are sterile. While the conjunctiva is protected by blood supply, the cornea is a vascular; thus, the types of organisms invading these tissues may be change. The intraocular tissues are relatively immune-privileged and can be infected by any organism that manages to enter the inside of the eye(Sharma, 2012).Bacteria, fungi, parasites and viruses, can all causesocular infections and each of theses may give a sings and symptoms of disease.Furthermore, the prevalence and distribution of spasific infectious organsims related with eye infections varies widely and is dependent on variety of factors (Gautamet al., 2019).
Fungal eye infection
Fungal eye infections are considered asignificant cause of loss of vision in some regions of the world, where they are the main causes of blindness especially in Asia. Fungal etiology of ocular illness is still a diagnostic and therapeutic problem for the ophthalmologist(Behera, 2018).The most common cause of eye infections is Candida species, that normaly develop in immunocompromised patients that have chronic underlying systemic disease.The second most commonis Aspergillus species, such as Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus terreus, Aspergillus glaucus, and Aspergillus nidulans.There have been many reported cases that reports other emerging pathogens, such as Fusarium spp. ,Penicillium spp., Pseudallescheria spp., Cryptococcus spp., dimorphic fungi (Histoplasma capsulatum, Blastomyces dermatitis, Sporothrix schenckii, and Coccidioides immitis)(Trofaet al., 2008 ; Spadea, and Giannico, 2019).
Methods
Two hundred eye swabs samples were collected in Wasit province, Iraq 2020. The samples were collected from different ages and included both genders. Initial identification of each sample was performed by culture on Sabouraud dextrose agar at 30°C for at least ten days and direct microscopy using lactophenole cotton blue staining. Molds identification according to Colony morphology (color and consistency) and Microscopic characteristics (micro conidia and macro conidia, their size, arrangement and shape). Yeast identification was made by VITEK 2 system method which is a fully automated microbiology identification system.
A B A B
Figure 1: Aspergillus spp. grown on SDA Figure 2 : Microscopic feature of Aspergillus
A: A.flavus, B:terrus, C: A. niger, D: A. fumigatus. Spp. stained with Lactophenol cotton blue A:A.flavus, B:terrus, C:A.niger, D:A. fumigatus.
Figure 3: Microscopic feature ofFigure 4: Cryptococcus laurentii A: Alternaria spp.and B:Penicillium spp.
Figure5: Candidaspp. grown on SDA.
A: Candidaciferrii, B: Candida glumornidii
Yeasts identified by the VITEK 2 system allows the identification of medically important yeasts and yeast-like organisms in 15 hrs due to a sensitive fluorescence-based technology and enable a result to be generated without the necessity for morphological testing(Grafet al., 2000).
However, in this study the results included two genera Candida and Cryptococcus.
C D C D
A B
A B
Figure 6: Identification of Cryptococcus laurentii by vitek2 system
Figure 7: Identification of Candida ciferrii by vitek2 system
Figure 8: Identification of Candida glumornidii by vitek2 system Data Analysis
Data entered and managed by using SPSS V 0.25 for Windows. Descriptive statistics (frequencies, mean ± standard deviation and with tables and graphs) and inferential statistics (Chi-Square Test) were used. P-value of ≤ 0.05 considered statistically significant.
Results
A sample of 200 participants involved in this study to isolate of fungal pathogens from patients with eye infections. The sample mean ±SD of age was 45.2±20.8 years (ranged 1-88 yrs.), 63.5%
of them aged ≥40 years old. Female to male ratio was 1.1:1 ; as outlined in the table (1).
Table (1): Demographic description of study sample (N= 200).
Variables Frequency %
Gender males 95 47
females 105 53
Age (years)
1-9 10 5
10-19 13 6.5
20-29 29 14.5
30-39 21 10.5
40-49 33 16.5
50-59 37 18.5
60-69 27 13.5
70-79 20 10
Tobacco smoking prevalent in 32 person (16%) of sample, figure (9). Nearly one-half of sample (104) present with Blood pressure, and 68 (34%) of them present with diabetes mellitus, figure (10).
Figure9: Distribution of tobacco smoking.
.
Figure10:Distribution according topresent/ absent of chronicdiseases
During the current study period, laboratory investigations indicated that 29 (15%) of the total samples diagnosed with a fungus species, 13 (45%) of them present with Aspergillusflavus followed by five cases (5) 17% with Aspergillusterrus , four cases (4) 14% with Aspergillusfumigatus, two cases (2) 7% with Aspergillus niger and one cases (1) 4% of each Penicillium spp. and Alternaria spp. , (1) 3% for each Candidaguilliermondii , Candidaciferrii and Cryptococcuslaurentii as outlined in the figure (11).
[VALUE]
(16)
[VALUE]
(84)
0 50 100 150 200
Smokers Non-smokers
FREQUENCY (%)
TOBACCOSMOKING
[VALUE]
(52)
[VALUE]
(34) [VALUE]
(48)
[VALUE]
(66)
0 20 40 60 80 100 120 140
Hypertension Diabetes Mellitus
FREQUENCY (%)
CHRONIC DISEASES
present
Figure (11): Distribution of fungus species.
Table (2): Relation of culture result with demographic characteristics, tobacco smoking and chronic diseases.
Factors
Culture result Statistic Positive
(n=29)
Negative (n=171)
Chi-Square
Test Sig.
Gender males (n=95) 20 (21%) 75 (79%)
6.267 0.012 females (n=105) 9 (9%) 96 (91%)
Age groups <40 yrs. (n=73) 9 (12%) 64 (88%)
0.437 0.509
≥40 yrs. (n=127) 20 (16%) 107 (84%) Tobacco
smoking
smoker (n=32) 10 (31%) 22 (69%)
8.621 0.006*
non-smoker (n=168) 19 (11%) 149 (89%) Type 2 Diabetes
mellitus
present (n=68) 18 (26%) 50 (74%)
11.909 0.001 absent (n=132) 11 (8%) 121 (92%)
Blood pressure present (n=104) 12 (12%) 92 (88%)
1.533 0.216 absent (n=96) 17 (18%) 79 (82%)
Discussions
According to the results there was statistically significant association between gender and the culture results of fungus (P 0.012), the fungus spp. detected in males two fold than in females this result was agreed with Thomas, (2003). The effects of smoking on the eye and ophthalmic diseases was first reported in the late 1970s. Current smokers may develop the disease about 10 years earlier than nonsmokers Zisimopoulos et al., (2021).the relation of healthy risky behavior with fungus culture data analysis indicated that there was a significant association between tobacco smoking and detection of fungus (P 0.006); fungus spp. identified in smoker patients three fold than in non-smokers this was agreed with Nita and Grzybowski,(2017),which found
Aspergillus flavus
45%
Aspergillus terreus
17%
Aspergillus fumigatus
14%
Aspergillus niger 7%
Alternaria 4%
Candida ciferrii 3%
Candida guilliermondii
3%
Cryptococcus laurentii
3%
Penicillium 4%
0%
Frequency
infection. Diabetes increases the risk of infection and damages multiple organs, which may affect the ability of protection against variety of pathogens. Poor glycemic control and chronic diabetes mellitus cause several complications like micro- and macrovascular complications, diabetic foot ulcers, eye infection, nephritis, and nerve infections, which are responsible for high morbidity and mortality Pearce et al., (2019).In addition, the results in the table (2) show that infection with fungus species associated with present of type 2 diabetes mellitus in which patients with type 2 diabetes mellitus have fungus infections three times more than in patients without diabetes (P 0.001) this result agree with Saud et al., (2020), who found that hyperglycemic state causes immune dysfunction which leads to local and systemic infection due to overgrowth of microflora and causes an opportunistic infection. However, there was no statistically significant association between the culture results of fungus and Blood pressure (P 0.216).
Conclusion
According to the results of the current study, we conclude that : Aspergillus spp. was reported as a high incidence in patients with eye infections, the relation between the fungal infections by any species was increased with diabetic patients, smoking has an effect with fungal eye infection and the blood pressure has no effect on fungal eye infection.
Limitations and Future Studies
Fungal eye infection should be put in mind by clinician inspite of its occurrence in low incidence because of their complications such as loss of vision if not successfully treated, studying of the predisposing factors those affecting the incidence of fungal eye infection and finally studying fungal eye infections on other people groups such as (pregnancy, AIDS, thalassemic patients,burns patients, dialysis patients and patients with hematological malignancies).
Acknowledgement
The authors would like to express their gratitude to the privit clinics of eye in Wasit province, Al-Zahraa teaching hospital staff, Al-Kut hospital staff for providing essential research facilities.
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