Shafaq haitham abbas *1, Shahad Haider Hanow2, Mohammad J. Al-jassani
1Department of Microbiology , College of Science , AlKarkh University ,Baghdad /Iraq
2Department, College, Al-turath University, Baghdad /Iraq
*Corresponding Author: .. shafaqhaitham121@gmail.com
Abstract
Background: Long-term metabolic disease known as diabetes mellitus (DM) is characterized by persistent hyperglycemia. Depending on the systemic infection or local, DM patients may be more susceptible to Candida spp.
Objective: Thus, the goal of the current investigation was to identify novel traits in the distribution, prevalence, and susceptibility to antifungals of Candida spp. isolated from patients with diabetes. We are able to evaluate the shift in the isolated Candida spp.’s antifungal sensitivity patterns thanks to the investigations.
Methods: During this study, 36 sample were collected from diabetic patient. On CHROMagar, Candida yeast was isolated and identified. Antibiotic Sensitivity was performed using antibiotic fluconazole, nystatin, itraconazole, amphotericin B,clotrimazole, ketoconazole
Results : A total of 17 sample showed no culture for any yeast . 19 sample were positive for yeast Growth. Three different Candida were identified. At 75%, Candida albicans, sometimes known as green colonies, was the most common species.
Conclusions: The current study that there is an association between clinical and microbiological findings like the isolation of Candida among Diabetes mellitus patients which might be causal. Candida species identification with the in vitro antifungal susceptibility pattern is essential to choose the appropriate antifungal drug and to predict the outcome of therapy.
Keywords: Candida,Diabetics,Identification
Introduction
Diabetes mellitus (DM) is a chronic metabolic disease characterized by hyperglycemia that does not go away. It could be brought on by resistance to insulin peripheral activity, insufficient insulin secretion, or both. Depending on whether DM patients have a systemic or local infection, there are a number of factors that contribute to a higher predisposition for candida spp. (Jha et al., 2021).
Candida species are a primary cause of Candidaemia and were previously thought to be culture pollutants (Mohammadi et al., 2016). It has long been known that patients with diabetes mellitus frequently have Candida infections (Willis et al., 1999). Numerous studies have demonstrated that people with diabetes had a higher prevalence of yeasts than healthy, normal people (Ferlay et al., 2015).
However, non-diabetics can also contract candidiasis if their immune systems are temporarily weakened by stress, alcohol, or HIV in the case of Hepatitis B patients (Fair et al., 2014). Additionally, some individuals with candidiasis are also taking antibiotics, as these drugs destroy microorganisms and allow Candida spp. to take their place (Sarah et al., 2017). Candida infection of the mouth and throat treated with a reputable antifungal medication and ongoing patient monitoring. The following symptoms could be present: lesion in the mouth that resembles cheese, as well as white, creamy soft patches within the cheeks and on the tongue. on the throat, discomfort and trouble swallowing, esophageal inflammation, and more symptoms (Jayachandran et al., 2016).
Currently, certain strains of Candida have developed resistance to the azole class of medications, particularly in those with diabetes (Delsing et al., 2021( Drug resistance in diabetic individuals is emerging as a significant cause for worry.
As a result, it is now crucial to periodically monitor the antifungal susceptibility pattern of the most common species of Candida. Thus, the goal of the current investigation was to identify novel traits in the distribution, prevalence, and susceptibility to antifungals of Candida spp. isolated from patients with diabetes. We can evaluate the shift in the isolated Candida spp.’s antifungal sensitivity patterns thanks to the investigations.
Material and Methods
Sample collection
Thirty six samples of lips and buccal lining samples were collected from Diabetic patient in Baghdad Iraq, 42 % the patients suffer from two types of diabetic. The sample were collected in Amis Transport medium (Himedia, India) and kept at 4°C until the tests were conducted.
Oral yeast culture and isolation
Swabs were cultured directly on CHROMagar (Himedia,India) to identify yeast and ensure culture purity according to the color of the colonies . The plates were incubated for twenty-four hours at 37°C.
Test of sensitivity for isolated yeast
The yeast isolates’ sensitivity was evaluated using different types of commonly used antifungal agents: Fluconazole, amphotericin B, Nystatin, Ketoconazole, Voriconazole, and Clotrimazole and as follows:
A single colony from each consecutive plate was cultured in a 3ml Molar Hinton broth over night at 37°C On a Molar Hinton agar dish, 1 ml of yeast solution (0.5 McFarland) was spread out, and it was left for 1 minute. Discs of antibiotics were placed on the culture . For a whole day, the dishes were incubated at 37° C. Every tablet containing diluted extracts had its inhibitory diameter measured, and it was then compared to the other tablets, which contained antibiotics and preparations for oral and dental hygiene.
A single colony from each consecutive plate was cultured in a 3ml Molar Hinton broth over night at 37°C On a Molar Hinton agar dish, 1 ml of yeast solution (0.5 McFarland) was spread out, and it was left for 1 minute. Discs of antibiotics were placed on the culture . For a whole day, the dishes were incubated at 37° C. Every tablet containing diluted extracts had its inhibitory diameter measured, and it was then compared to the other tablets, which contained antibiotics and preparations for oral and dental hygiene.
Results and Discussion
A total of 17 sample showed no culture for any yeast . 19 sample were positive for yeast Growth . Nine of them with very few colonies (green) while 10 showed havy growth some of them are mixed cultures Table (1) and Figure (1). different Candida spp were isolated . Candida albicans (green colonies) are the most common, accounting for 75% of the fungal isolates, this concurs with (Willis et al.1999).
Selecting an antifungal medication is made more difficult by the shift in the etiology of oral candidiasis from Candida albicans, the species that is frequently encountered, to non-Candida albicans such C. glarata and C. krusei, the more intrinsically drug-resistant species (Mäkinen et al. 2018).
Sensitivity test
The outcomes of the sensitivity test for the yeast isolates during investigation are displayed in Table (2). The outcomes demonstrated that all samples were moderately sensitive to Ketoconazole, Voriconazole, and Clotrimazole with very weak sensitivity to amphotericin B Table (2) and Figure (2A). Only one sample (28) with a blend culture of C. tropicalis and C. albicans showed resistance to all tested antibiotics Figure (2B). One of the first-line medications for treating candidiasis in the mouth in cancer patients is fluconazole (Delsing et al., 2021; Mäkinen et al., 2018). The standard treatment for extensive Candida infections is amphotericin B. Fungal infections have become more common in those with compromised immune systems, including diabetic patients receiving radiation therapy and chemotherapy (Delsing et al., 2021). The dry mouth mucosa caused by cytotoxic therapy makes it easier for several bacteria to infect patients. According to studies, the two stages of the development of candidiasis are colonization and then invasion of the epithelial layer (Delsing et al.2021, Bombeccaria et al.2016). Once colonization is established, the epithelial layer might be invaded due to compromised cellular immunity. Chemotherapy, radiation, and neutropenia will cause mucositis, which will rupture the mucosa and allow Candida to invade deeper (Delsing et al., 2021). Antifungal resistance in Candida species is a major concern, especially in diabetic patients, as these drug-resistant isolates have the potential to spread to deeper tissues and cause infection. Because of the high frequency of Candida in the mouths and throats of cancer patients receiving chemotherapy or radiation therapy, systematic monthly surveillance of fungus infections is necessary to identify patterns of antifungal resistance.
Extensive chemotherapy might develop drug-resistant microbes. Add to that, the presence of such microbes might participate in insitue Diabetic initiation and development(Jayachandran et al.2016). According to multiple studies (Mäkinen et al.2018, Bombeccaria et al.2016), the incidence rate of Candida infection in oral squamous cell carcinomas of the head and neck region ranges from 25% to 74.7%. The presence of Candida in these tumors is linked to a poorer prognosis.
Table 1: Identified yeast species on CHROMagar medium
Code
Species
Color on CHROMagar
2
Candida albicans
Green
9
C. albicans
Green
14
C. albicans
Green
16
C. albicans
Green
23
C. albicans
Green
25
C. albicans
Green
30
C. albicans
Green
31
Candida albicans
Green
32
C. albicans
Green
3
C. krusei
purple
7
C. krusei
purple
15
C. krusei
purple
18
C. krusei
purple
24
C. krusei
purple
26
C. krusei
purple
27
C. krusei
purple
29
C. krusei
purple
8
C. glabrata
pink
21
C. glabrata
pink
Figure 1: On CHROMagar medium, isolated yeast (A: C. Krusei, B: C. glabrata, and C: C. albicans) is displayed
Table 2: Evaluation of the isolated yeast’s sensitivity
NO.
Antibiotic
Mean Inhibition zone diameter (mm)
1
Amphotericin B (AMB)
7
2
Ketoconazole (KCA)
14
3
Clotrimazole (CLO)
17
4
Nystatin (NY)
–
5
Itraconazole
7
6
Fluconozol (FLU)
_
Figure 2: Sensitivity test results
Conclusions
A prevalent fungi infection among diabetic people receiving chemotherapy is oral candidiasis. In some cases, the correlation between clinical and microbiological results can assist in linking specific symptoms to the isolation of Candida in diabetic patients; nevertheless, these correlations may be causative. The antifungal susceptibility pattern of both Candida albicans and non-Candida albicans is same. Selecting the right antifungal medication and forecasting treatment results need species-level identification using an in vitro antifungal resistance pattern.
Acknowledgment
We thanks to all people who donated their swap samples for the approval of analysis and to all the laboratory staff of the medical technician at hospital in Wasit and department of medical laboratory ,Middle technical university.
Conflicts of Interest
There are no competing interests.
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