Prevalence of antibodies in Iraqi Urinary Tract Infection patients using radial immunodiffusion (RID) assay

Saja Mohammed Mohsen* Anas Wisam malik DOI. 10.21931/RB/2021.06.04.23 Abstract: Candida has different types that could cause bloodstream infections. A total number of 150 samples were collected from candidemia patients and examined. The Candida spp. Species isolated from blood samples were analysed. These were identified by culturing the species using different media, namely the chromogenic agar test. Then, the virulence factors of all samples were tested. The Candida glabrata isolates were tested with six commercial antifungal drugs. C. glabrata 67 (44.6%), C. albicans 34 (22.6%), C. krusei 18 (12%), C. tropicalis 17 (11.3%), and C. parasilosis 14 (9.3%). the production of phospholipase ranged between 0.63-0.99 mm. It was found that 96% of the species showed phospholipase activity in aerobic conditions. The protease activities of Candida spp. Isolates were experimentally tested by area of inhibition around the colonies, where 59.3% had the double (++) protease activity, 31.4% with (+) grade, and 9.3% had (–) grade or clear zone around the colony. The hemolytic capacity ranged from 0.69-0.89 in the optimum aerobic environments. Finally, 38.33% of the isolated Candida spp. were positive and 61.67% negative for biofilm formation. Out of the total positive Candida spp. for biofilm formation, 21.73% were strong biofilm producers, and 78.27% were weak. Minimum fungicidal concentration (MFC) of Fluconazole for C. glabrata isolates was not appropriate (NA) due to the occurrence of low inhibition tested for species. Micafungin exhibited the lowest fungicidal activity against C. glabrata ranging from 0.03 0.125, while Fluconazole showed the highest.


Introduction
Urinary tract infections (UTIs) are common bacterial infections, with nearly more than one hundred and fifty million individuals infected yearly 1 . In the United States, estimated 10.5 million clinic cases with UTI symptoms and 2-3 million cases visit the emergency department [2][3][4] .
The urinary tract includes kidneys, ureters, bladder, and urethra. Several natural factors are secret with urine from the urinary tract for protecting against all infection agents; also, the anatomical and mechanical barriers have a significant role, such as the glycoprotein plaque uroplakins 5 and a layer of hydrated mucus 6 . Furthermore, the urinary tract lining is made by immune cells and epithelial cells, which have an important role in protecting against bacterial infection and preventing bacteria from getting into the urinary tract 6 . UTIs are most common in females more than males, and the incidence rate has increased during the last (30) years 6 .
UTIs increase in the elderly due to the immune-compromised individuals, and the urinary catheter has an essential role in increasing the percentage of infection 7 .
Immunity has two types innate immunity and adaptive immunity [8][9][10] . The individual with innate immunity has resistance against the infection. Innate immunity includes many chemical and cellular components. The infection gets in and spreads, but the person with high immunity has resistance against the bacteria 11 . Adaptive immunity has two types of humoral immunity, and cellular immunity includes all the naturally acquired immunity due to infection vaccination [12][13][14][15] .
Features likely to contribute to disease of the human immune system involve the responses to antigens (B-and T-cell features, PMNs), the efficiency of bacterial destruction (e.g., lysozyme, complement), and types of antimicrobial substances produced (e.g., immunoglobulins, Cytokines) 16 . Immunoglobulins are a group of serum proteins with a crucial antimicrobial activity; IgM represents an indicator of recent infection, IgG levels increased in chronic infections 17 .

Materials and methods
This study was conducted around UTI Iraqi patients during the period from November 2019 to April 2020. This study included 40 patients and 20 healthy controls.

Samples collection
Serum samples were collected from the patient in Al-Khalis General Hospital and National Laboratories in Al-Khalis District / Diyala Governorate (Iraq) from November 2019 to April 2020. After examining their urine samples by the microscope, they were divided into two groups first is patients with UTI, and the other group is healthy who does not suffer from UTI; this group is considered the control for comparison with the first group.

Estimation of antibodies
Using radial immunodiffusion test to detect levels of antibodies in serum is done by using (LTA, Italia). Normal values of antibodies were IgG (800-1800 mg/dl) and IgM (60-280 mg/dl).

Statistical Analysis
The Statistical Analysis System (SPSS) program was used to detect the effect of different factors in study parameters least significant difference, using T-test for comparing between the means value and the using of the Chi-Square test for comparing between the percentage.

Results and discussion
The patient group involved 40 patients, 8 males 20% and 32 females 80%, with a mean age of 40.60 ± 2.295 years. The healthy control group consisted of 20 subjects, 11 males ( 55%) and 9 females (45%), with a mean age of 36.80 ± 3.441 years, as seen in tables 1 and 2 explain the distribution of the two study groups according to gender and age.
Corresponding author: sajamohammad@mtu.edu.iq Tables 1 and 2 show UTI infections occur in females more than males; the mean age is 40.60 ± 2.295 years. The higher incidence of UTI in females than males might be due to a variety of factors, such as the proximity of the female urethral meatus to the anus 18 . Also, most school girls are avoiding urinable in school, which is accompanied by urinary infection 19 . Alternatively, vaginal flora has a vital role vaginal infection with coliforms which leads to urinary tract infection 20 . The female anatomic feature also contributes to higher prevalence among the female subject. This finding is agreed with earlier studies [21][22] .

Results in
There was a significant increase in the mean concentration of IgG in sera of patients (854.82±138.79) mg/dl compared with the mean concentration of IgG in sera of the control group (616.4 ± 67.65) mg/dl. These results show high significance between two groups UTI patients and control group according to IgG while non-significant to IgM show table 3. This finding is agreed with AlSaadawi and Alkhaled (2015) 23 . Elevation in serum concentration of IgG in patients suffering from UTI may be the reason for the significant increases in the concentration of complement components in patients suffering from UTI24. Higher values IgG undergo increases to protect and combat infection 24 . Table 4 shows the non-significant difference in gender distribution between two groups according to IgG

Conclusions
We conclude that UTI infection occurs in females more than males; the mean age is 40 years. In all patients with UTI compared to control, the IgG serum level increases and IgM serum level decreases. Serum IgG and IgM showed high significance between two groups of UTI patients and the control group according to IgG while IgM showed non-significant and serum IgG and IgM showed no significant difference in UTI patients and RBC groups.
Bibliographic references Table 5. Effect of present RBC on IgG and IgM prevalence in UTI patients.