INTRODUCTION:

Generally, infection with a single cell Toxoplasma gondii parasite cause toxoplasmosis, this parasite can be found in the entire world, this infection can threat the life because it cause weakness in the immunity system of the human being¹, T cells may involved with this infection and decrease the immunity system².

Vitamin D play an important role in regulating and the metabolism of phosphorous and calcium as well as it is consider as modulator to the immunity, it may also prevent the proliferative particularly 7-25-OH2D³, also it may participate in prevent diseases via decreasing IL2, however deficiency in vitamin D is rising all over the world⁴, it reach around 70% of population in Iran⁵, and because VD vitamin D affect the biological activities in human like; ergocalciferol D2 and cholecalciferol D3, it is important to keep it in a certain concentration in the sera, and the perfect amount of it in adults must be not less than 20ng/mL^6,7.

Mainly, it is well known that VD keep bones healthy and conserve them, however recent studies shown the association between the level of VD in sera and a lot of diseases such as many types of cancer⁸, including prostate cancer, breast cancer, colon cancer, and all the consequences in health of the human^9,10.

Many studies were carried out to find the correlation between deficiency in VD and pregnant women, such as a study carried out by Mulligan et al., 2010¹¹, the study discuss the impact of the deficiency in VD and the metabolism and lactation in pregnant women, in 2019 Fakhrieh Kashan and her coworkers¹² conclude that infection with gondii parasite has relation with deficiency in VD which obviously cause toxoplasmosis.

Another study carried out by Zhang et al., 2020¹³ found that there is a correlation between toxoplasmosis and the lake in VD in childbearing age women.

In current research was carried out to evaluate the association between the lake in vitamin D and toxoplasmosis seropositivity in the women’s sera in the age of childbearing via ELISA.

MATERIALS AND METHODS:

A total of 56 women their ages range between (17-45) years. The 25-OH Vtamin D (total) was determined using ELISA in the plasma and serum (Monobind Inc.USA, ACCUBind, ELISA Microwells) 25-OH Vitamin D –Direct Test system product Code: 9425-300.

Test Procedure for vitamin D detection:

All references, reagents controls and calibrators must be at RT (room temperature) around 23.5±3.5°C). the wells of the microplate were set-up for every serum, calibrator, reference, control and the serum of the patients were assayed two times. The unused microwells strips were replaced sealed and stored at 2-8°C.

1. 25µl of the appropriate extracted 25OH Vit D calibrators, control, or the specimen were pipetted in the wells.

2. Add to the well 100µl of the 25-OH Vit D Releasing Agent.

3. Then mixing the microplate for 20-30 seconds until homogenous.

4. Then cover and incubate the microplate at RT for 30 minutes.

5. Then discard the microplate content by aspiration or decantation.

6. 350µl of wash buffer was used to wash the microplate for 2 to 3 times.

7. Then, to the wells, add 100µl of 25-OH Vit D Enzyme Reagent.

8. Then cover and incubate the microplate at RT.

9. Then discard the microplate contents by aspiration or decantation.

10. Repeat step 6 and 9.

11. Add to all the wells 100µl of substrate reagent.

12. Incubate the plate for 20 minutes at RT.

13. To each well, add 50µl of stop solution was and mixed for 15-20 seconds.

14. Then absorbance was taken at 450nm.

RESULTS:

Fifty six females were studied for toxoplasmosis following immunoglobulin estimation. Two females had positive IgM levels, 41 females had positive IgG levels and one of the females had both IgG and IgM positive. The total was 44 positive for toxoplasmosis. The following table presents the demographic characteristics of the studied females.

Statistical analyses were performed using SPSS (version 20.0 for windows, SPSS, Chicago, IL, USA). Data were presented as mean±SD, range, median for age, the P value was significant at P<0.05.

The results shows that 88.6% of housewives women had toxoplasmosis, while only 11.4% of worker women did not have toxoplasmosis, also the results indicates that there was no relationship between presence or absence toxoplasmosis and job P=0.0635. Butthe results revealed that toxoplasmosis related to menstrual regular cycle P = 0.043,since there were 44(100%) women had regular cycle and had toxoplasmosis, while there were only 10(83.3%)women did not have toxoplasmosis and had regular cycle and 2(16.7%) women did not have toxoplasmosis and had irregular menstrual cycle.

The results also showed that there were 4(9.1%) women infected with toxoplasmosis and had changes in the menstrual cycle, and there were 3(25%) women did not have toxoplasmosis and did not have changes in the menstrual cycle, toxoplasmosis was not related to menstrual changes P=0.16. In addition the results showed that there were 21(47.7%) women infected with toxoplasmosis and had abortion, while there were 4(33.3%) women did not have toxoplasmosis and had abortion, there was no relation between toxoplasmosis and abortion P=0.516.

In addition the mean±SD of the number of abortion in women infected with toxoplasmosis was 0.66± 0.75(0-3). While in women not infected with toxoplasmosis mean±SD was 0.58± 1(0-3), the number of abortions wasn’t a noticeable different from the control group P=0.245. The mean±SD of age of women infected with toxoplasmosis was 27.9±6.9(17- 45), and in women not infected with toxoplasmosis the mean ±SD was 25± 6.5(17-43). The age was not significantly different with the control group females (P=0.221). The mean±SD of number of abortion in female infected with toxoplasmosis was 0.66± 0.75(0-3), while in women not infected with toxoplasmosis was 0.85±1(0-3).

The number of abortions was not significant different (P=0.773). The mean±SD of number of pregnancies in women infected with toxoplasmosis was 1.11±0.97(0-4), while in women not infected with toxoplasmosis was 0.75±0.87(0-3). There was no significant differences between the number of pregnancies and toxoplasmosis (P=0.45).

The present study showed that there were 44 women had toxoplasmosis and had deficiency of vitamin D3 levels, since their mean±SD was 5.42±4.59 (Table -1), while there were 12 women did not have toxoplasmosis and had deficiency of vitamin D3 levels mean ±SD was 6.34±9.86. There wasn’t obvious differences in the concentration of vitamin D3 between patients group and control group P=0.643.

Table (1) showed that there were 43(97.7%) women had toxoplasmosis and had 11(91.7%) women did not have toxoplasmosis and had deficiency in the levels of vitamin D3 in their sera. However almost all females had a deficient levels of vitamin D3 with (above 20ng/ml) in the control group with a percentage of 8.3% compared with toxoplasmosis females (2.3%), but the relation was not obvious (P=0.386).

Table (2) showed that there were 25(56.8%) women had very severe deficiency in vitamin D3 concentrations, and there were 14(31.8%) women had toxoplasmosis and had severe deficiency of vitamin D3 concentrations ,there was only 1(2.3%) women had toxoplasmosis and had suboptimal levels of vitamin D3. There was no obvious relation between vitamin D3 levels and toxoplasmosis (P= 0.283) Figure 1.

Table 2: frequency of deficiency in vitamin D among women infected or not infected with toxoplasmosis

		Toxoplasmosis
		+ve		-ve
		Count	%	Count	%
Vit D3	Very severe deficiency	25	56.8%	7	58.3%
	Severe deficiency	14	31.8%	4	33.3%
	Deficiency	4	9.1%	0	0.0%
	Sub optimal	1	2.3%	0	0.0%
	Optimal	0	0.0%	1	8.3%

Figure 1: vitamin D3 concentrations (mean value) in women infected or not infected with toxoplasmosis

DISCUSSION:

The current study shows that 88.6% of housewives women had toxoplasmosis, while only 11.4% of worker women did not have toxoplasmosis, these results in agreement with reported by³⁶who mentioned the high incidence of toxoplasmosis in housewives women than that in worker women, since the results of the present study included high number of housewives women in comparing to the number of employees women. The reason for the increased exposure of housewives to cat disease more than working women is probably due to the fact that housewife does a lot of housework, such as handling with contaminated meat, cats, vegetables and cleaning the garden of the house from cat litter, which contains many infective stages of the T. gondii. Also the results of the present study showed that mean±SD of age of women infected with toxoplasmosis was 27.9± 6.9(17-45), while the mean±SD was 25.2±6.5(17-43) in women not infected with toxoplasmosis, there was no obvious variation between two groups P= 0.221,these results in agreement with t, hat reported by¹⁴, that the incidence of toxoplasmosis infection is higher in warmer and humid climate and increases with age .The variability in the rates might be due to several factors, such as type of laboratory method, samples sizes, as well as the patients themselves^15,16,17.

Additionally, there were 42(75%) women had toxoplasmosis and had positive results of IgG Abs of toxoplasmosis, while there were 12(21.4%)women had negative results of IgG Abs of T.gondii, while there were only 2(3.57%) women had positive results of IgMAbs of T.gondii .These results incompatible with that reported by^18,19that 33% of the worlds´ women who were infected with T.gondii stay asymptomatic due to the response of immune system to prevent parasite action to cause illness, these results in agreement with that reported by^20,21that the elevation of Toxo-IgG Abs 49.01% was due to that women who got T.gondii previously, and they already have high level demand of anti Toxo-IgG Abs they exclude seroconversion and to assure protection^22,23.

Also the results were in agreement with that reported by¹⁹, that IgG and IgM +ve (Anti-T. gondii IgG antibody was positive in 17.14% of usual vitamin D group and 28.57% in deficiency group (P≤0.05), these differences was obvious and it is an evidence that Toxoplasma infection is linked with deficiency in vitamin D.

Table (1) showed that there were 43(97.7%) had toxoplasmosis and had deficient in vitamin D3 concentration, while in control group there was 11(91.7%)women did not have toxoplasmosis and had deficiency of vitamin D 3 concentrations and there was 1(2.3%) women had toxoplasmosis and had optimal levels of vitamin D3 concentrations, and 1(8.3%) women did not had toxoplasmosis and had optimal levels of vitamin D3. There was no significant difference in the concentration of vitamin D3 with the control group (P=0.643); however almost all females had a deficient level of vitamin D3 with a slightly higher optimal vitamin D3 level (above 20ng/ml) in the control with a percentage of 8.3% compared with toxoplasmosis females (2.3%) but the relation was not significant (P=0.386), this result in a conflict with findings of^24,25.

Also, in table 1, the mean±SD vitamin D3 level in women had toxoplasmosis was (5.42±4.59), while in control group the mean±SD was (6.43±9.8), which indicates that there was no important differences in the concentration of VD3 with the control group (P=0.643). The results disagreement with that reported by^26,27that there was significant differences correlation between toxoplasmosis and vitaminD3 deficiency, and this finding was preliminary and prone women for getting osteoporosis. Illustration of the results of the present study that there was no correlation between vitamin D3 levels and toxoplasmosis may be due to the low number of sample size collected or may be due to the errors occurred during laboratory work to check for vitamin D levels, since it is sensitive kit and needs special conditions to work, or perhaps for a delay in reading in the spectrophotometer^28,29.

Table (2) showed that there were 25(56.8%) women infected with toxoplasmosis and had very deficiency in vitamin D levels, while there were 14(31.8%) women had sever deficiency in vitamin D levels, and infected with toxoplasmosis, while there was only 1(2.3%) women had toxoplasmosis and had suboptimal levels of vitamin D3.There was no significant relation between vitamin D3 levels and toxoplasmosis (P=0.283), the number of women had deficiency in vitamin D levels infected with toxoplasmosis was high than the number of women not infected with toxoplasmosis, these results in agreement with that reported by²¹that about one- third of people are born with anti-Toxoplasma antibodies worldwide, vitamin D deficiency rate was 63.93% among women in relation to toxoplasmosis, since IgG and IgM +ve (Anti-T. gondii IgG antibody) was in 17.14% (positive) in normal VD group and was 28.57% in deficient VD group (P≤0.05), these differences was obvious and evidence that Toxoplasma infection is associated with deficiency in VD. The association between seropositive toxoplasmosis with, disorder in VD, minerals, and bone markers was significant.

Furthermore the deficiency in VD in Iraqi population could be related to the following:

· No enough VD in their diet.

· Low exposure to sunlight.

· Health issues (the body cannot benefit from VD) such as:

· Liver diseases;

· Renal disease;

· Gastrointestinal disorder and others.

On the other hand, the role of T. gondii on the bones remains unclear and difficult to understand (obscure), and the question that appear how can parasite develop and affect a rigid and solid tissue like bone? Therefore, this study recommends more in vitro and in vivo studies to discover the impact of parasite on the bone infection and collect more data and more information and the role of immune system on this infection. The health education of women may lower the risk of this infection and lower its possibility.

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Received on 02.11.2021 Modified on 23.12.2021

Research J. Pharm. and Tech 2022; 15(9):4073-4077.

DOI: 10.52711/0974-360X.2022.00683

	Toxoplasmosis
	+ve (N=44)				-ve (N=12)
	Mean	SD	Min	Max	Mean	SD	Min	Max
Vit D3 ng/ml P=0.643	5.42	4.59	0.53	24.40	6.34	9.86	0.01	36.60
	Count		%		Count		%
Vit D3 group
Deficient	43		97.7		11		91.7
Optimal	1		2.3		1		8.3