Distribution of Blood Groups among Patients with Oral Candidiasis and their Secretor Status

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1 ORIGINAL ARTICLE Distribution of Blood Groups among Patients with Oral Candidiasis and their Secretor Status S Jeelani, GS Asokan, M Manjunath, Josy Jacob ABSTRACT Introduction: Research in the field of predictive and preventive medicine is an interesting field predicting the future resistance or susceptibility to diseases. Many parameters play a significant role in this aspect. Secretor status and blood groups are one such data in predicting future health. This paper is a reflection of a study in oral candidiasis patients using a simple blood and saliva test to study the relationship between the above parameters and the disease status. Objectives: To determine the distribution of ABO Blood groups, secretor status in oral candidiasis patients and to compare them with healthy individuals. Materials and methods: This study was carried out in two groups of 30 subjects in each group, namely control group (normal healthy individuals) and a group consisting of patients with oral candidiasis who were subjected to blood grouping and a test to assess the secretor and nonsecretor status by slide agglutination method and hemagglutination inhibition principle respectively. The statistical analysis was done using chi squared test. The degree of freedom, p-value and the statistical significance were assessed. Results: The results were less statistically significant in the secretors and nonsecretors of the controls and study groups according to different blood groups and statistically significant when comparing between the total number of secretors and nonsecretors in controls and patients with oral candidiasis. Also the distribution of secretor status in control according to their blood groups is statistically significant unlike in patients with oral candidiasis. Conclusion: Nonsecretors are much more like likely to be carriers of Candida species and to have problems with persistent Candida infections and blood group O are the most affected. Keywords: Secretor status, Blood groups, Oral candidiasis. How to cite this article: Jeelani S, Asokan GS, Manjunath M, Jacob J. Distribution of Blood Groups among Patients with Oral Candidiasis and their Secretor Status. J Indian Acad Oral Med Radiol 2013;25(4): Source of support: Nil Conflict of interest: None INTRODUCTION Despite the advances in science and the exceptional knowledge of elite scientists in the contribution to development in technology yet the counterattack of nature in the form of diseases in the most unpredictable manner is an everlasting challenge. Research in the field of predictive and preventive medicine is an interesting and exciting field predicting the future resistance or susceptibility to diseases. Many parameters play a significant role in this aspect. 256 Secretor status and blood groups are one such data in predicting future health. This paper is a reflection of a studybudding effort and a holistic attempt to explore the association of both the variables to health status of an individual. This study involves the variables blood groups and secretor status in oral candidiasis patients using a simple blood and saliva test to study the relationship between the parameters and the disease status. A blood type (also called a blood group) is a classification of blood based on the presence or absence of inherited antigenic substances on the surface of red blood cells. These antigens may be proteins, carbohydrates, glycoproteins or glycolipids depending on the blood groups system and some of these antigens are also present on the surface of other types of cells of various tissues. 1 A secretor is defined as a person who secretes the ABH antigens into body fluids and secretions like the saliva, tears, gastric mucus and this occurs in approximately 80% of population. The secretor status is represented by secretor gene. 2 The main aim and objective of this study is to determine the distribution of ABO blood groups, secretor status in oral candidiasis patients and to compare them with healthy individuals. MATERIALS AND METHODS This study was carried out in two groups of subjects to determine the distribution of blood groups and secretor status. The two groups consisted of a control group (normal healthy individuals) and a group consisting of patients with oral candidiasis with 30 subjects in each group. Oral candidiasis was diagnosed by cytological smear test using periodic acid schiff staining technique. 3 Both the groups were subjected to blood grouping and a test to assess the secretor and nonsecretor status. The blood groups were determined by slide agglutination method and the secretor status was assessed by employing the principle of hemagglutination inhibition principle. The statistical analysis was done using chi-squared test. The degree of freedom, p-value and the statistical significance were assessed. METHODOLOGY IN THE DETERMINATION OF BLOOD GROUP 4 Armamentarium: Glass slides, pasteur pipettes, applicator sticks, centrifuge, anti sera A, anti sera B, normal saline.

2 JIAOMR Specimen collection and preparation: About 1 ml of blood was collected in EDTA bulb and tested on same day of collection. The collected blood was stored at 2 to 8º till the time of testing. PROCEDURE Ten percent suspension of red blood cells was red blood cells were prepared in normal saline. On one half of the glass slide, place 1 drop of anti A blood grouping system. On the other half of the glass slide place 1 drop of anti B blood grouping system. Using a Pasteur pipette add 1 drop of cell suspension to each half of the slide, with separate applicator sticks mix each cell serum mixture well. Tilt the slide back and forth and observe for agglutination. Tests that show no agglutination within 2 minutes are considered negative. Peripheral drying or fibrin stands should not be interpreted as agglutination. Reaction Distribution of Blood Groups among Patients with Oral Candidiasis and their Secretor Status Interpretation Anti A Anti B Groups + O A O + B + + AB O O O O: No agglutination; +: agglutination Rh factor is determined using the same procedure. On a clean glass slide one drop of anti D serum is used. By using Pasteur pipette, one drop of whole blood is added to anti D sera. With the help of applicator stick cell-serum mixture mixed well. Slide is tilted back and forth and observed for agglutination. Tests that show no agglutination within 2 minutes are considered negative. DETERMINATION OF SECRETOR AND NON SECRETOR STATUS 5 The presences of blood group antigens in the saliva of the subjects of both the groups were tested as follows: Saliva Collection and Processing About 1 ml of unstimulated saliva was collected in a dry test tube and was placed in a boiling water bath for about 10 minutes to destroy the enzymes in saliva which otherwise would affect the activity of antigens and lead to false positive results of agglutination. After heating of the saliva, the tube was centrifuged at the speed of 3000 revolutions per minute for about 20 minutes. The supernatant fluid was separated and used for the secretor test. The test was carried out at room temperature employing hemagglutination inhibition technique introduced by Weiner and modified by Roy and Chatterjee in Principle of Hemagglutination Inhibition Technique If there is a presence of specific blood group substance in saliva and to it if corresponding specific antibody serum is added then the antibodies in that serum are inhibited by the antigen in the saliva. Now if red blood cells of the same group are added to this saliva-antiserum mixture there will be no agglutination due to the previous inhibition of the antiserum. Thus in case of Secretor there will be no agglutination. However, if the subject is nonsecretor then there will be no blood group specific substance in his saliva. So the anti serum added to it will not be inhibited and if red cells of this group are added to the saliva-antiserum mixture the antigens on these cells will react with the antibodies in the mixture and there will be agglutination reaction. Thus in case of nonsecretor there will be an agglutination reaction. Two percent suspensions of red cells were used for the test. Preparation of 2% Suspension of Red Cells First to pack the red cells the freshly collected blood was taken in a centrifuge tube and the tube was centrifuged for 15 minutes. The supernatant plasma was removed. The remaining packed cells were well mixed with sterile, isotonic saline which was taken in ample amount. This mixture was again centrifuged and the supernatant fluid was removed, again the packed cells were mixed with sterile, isotonic saline. The procedure was repeated atleast 4 times to get the red cells washed. After the last washing supernatant was removed along with upper layer of packed cells. The remaining volume of packed cells is taken as 100% and from it 2% suspension of red cells in sterile, isotonic saline was prepared. Hemagglutination Inhibition Technique The processed saliva was diluted 1:10 in cases of the subjects belonging to blood group A, B or AB and 1:5 in case of the subjects belonging to O group. All samples were tested as follows using 6 test tubes. Tube 1: 0.2 ml of diluted saliva ml of diluted anti A serum. Tube 2: 0.2 ml of diluted saliva ml of diluted anti B serum. Tube 3: 0.2 ml of diluted saliva ml of diluted anti A serum (control). Tube 4: 0.2 ml of diluted saliva ml of diluted anti B serum (control). Tube 5: 0.2 ml of diluted saliva ml of diluted anti H serum. Tube 6: 0.2 ml of normal saline ml of diluted anti H serum (control). Journal of Indian Academy of Oral Medicine and Radiology, October-December 2013;25(4):

3 The contents were well mixed and were allowed to stand for 15 minutes. Then 2% respective red cell suspension was added to the respective test tubes. The contents in the tubes were mixed well and the tubes were allowed to stand without disturbing them. All the tubes were incubated at room temperature for 2 hours. The results were read microscopically. The absence of agglutination reaction indicated the presence of antigen in saliva, i.e. secretor status, while presence of agglutination reaction indicated the absence of antigen in saliva, i.e. nonsecretor status. RESULTS AND OBSERVATIONS The study was carried out in 30 subjects with oral candidiasis which was diagnosed by cytological smear test using Periodic acid Schiff staining technique. All the patients were subjected to blood grouping and secretor status assessment by slide agglutination method and hemagglutination inhibition principle respectively. The data was subjected to chi squared test and degree of freedom, value and statistical significance was analyzed. The following observations were made as part of the study. OBSERVATIONS 1. The blood groups among the oral candidiasis patients was observed as 76.67% with O group; 13.33% with A group; 10% with B group and none were with AB group (Graph 1). 2. The secretors status among oral candidiasis patients was assessed as 30%-secretors and 70% nonsecretors (Graph 2). 3. The secretors status among oral candidiasis patients of different blood groups was observed as. Secretor status: 30.43% O group; 25%-A group; 33.33% B group; none with AB group. Nonsecretor status: 69.57% O group; 75% A group; 66.67% B group; none with AB group Chi-squared = 0.06; degree of freedom = 3; p-value = The test is statistically less significant (Graph 3). 4. The secretor status in control and in patients with oral candidiasis according to their blood groups was assessed as: Control group: O group: 33.33% secretors, 16.67% nonsecretors A group: 20% secretors, 3.33% nonsecretors B group: 20% secretors, 3.33% nonsecretors AB group: Nil secretors, 3.33% nonsecretors Chi-squared = 4.14; degree of freedom = 3; p-value = The difference is statistically significant: Study group: O group: 23.33% secretors, 53.33% nonsecretors A group: 3.33% secretors, 10% nonsecretors B group: 3.33% secretors, 6.67% nonsecretors AB group: nil secretors, nil nonsecretors Chi squared = 0.06; degree of freedom = 3; p-value = The difference is not statistically significant. DISCUSSION Research in the field of preventive medicine is an interesting area predicting the impact of pathologies on systemic health. Many parameters play a significant role in this aspect. Secretor status and blood groups are one such data in predicting future health. Several studies were conducted to explore the relationship between secretor status and overall health status. Systematic literature search revealed the following: Graph 1: Distribution of blood groups among the oral candidiasis patients 258

4 JIAOMR Distribution of Blood Groups among Patients with Oral Candidiasis and their Secretor Status The significance of secretor status on systemic health was highlighted by Grundbacher FJ who brought out the concept of defense in depth strategy and preclusive strategy which is explained as follows. The ABH nonsecretor status is associated with a defense in depth strategy which implies that let the invader invade in and attempt to destroy it internally. The preclusive strategy implies that wall out the invader and do not allow entrance in the first place. 6 The effect of secretor status on different systems of the body was found to influence various pathologies, such as myocardial infarction, 7 bronchial asthma, 8 gastroduodenal ulcer, 9 insulin dependent diabetes 10 and variety of autoimmune diseases. 11 The impact of secretor status on oral health was studied considering a common problem, such as dental caries wherein it was found that in all blood groups the average amount of cavities is lower for ABO secretors than nonsecretors. This difference is most significant for smooth surface areas of the teeth. Also, secretors of blood group A had the lowest number of cavities. 12 Thom SM et al focused on the correlation between secretor status and oral candidiasis and showed that blood group O nonsecretors are the most affected. One of the innate defenses against superficial infections by Candida species appears to be the ability of an individual to secrete the water soluble form of the ABO blood group antigens into the body fluids. The protective effect afforded by the secretor gene might be due to the ability of glycol compounds in the body fluids of secretors to inhibit adhesions (attachment lectins) on the surface of the yeast. In attachment studies, preincubation of blastospores with boiled secretor saliva significantly reduced their ability to bind to epithelial cells. ABH nonsecretor did not reduce the binding and often enhanced the numbers of attached yeasts. 13 The overall observation of the present study showed that the difference was less statistically significant in the secretors and nonsecretors of the controls and study groups, i.e. oral candidiasis according to different blood groups. Also the tests showed that the difference was statistically significant when comparing between the total number of Graph 2: Distribution of secretors status among oral candidiasis patients Graph 3: Distribution of secretors status in oral candidiasis patients of different blood groups Journal of Indian Academy of Oral Medicine and Radiology, October-December 2013;25(4):

5 secretors and nonsecretors in controls and patients with oral candidiasis and also the distribution of secretor status in control according to their blood groups unlike in patients with oral candidiasis in whom the difference was not statistically significant. CONCLUSION The significant findings of various workers are conflicting and no uniform association has been found between distribution of blood groups, secretor status and oral candidiasis although preponderance of one or other blood groups has been reported from time to time. Nonsecretors are much more like likely to be carriers of Candida species and to have problems with persistent Candida infections and blood group O are the most affected. To conclude since blood groups and secretor status have a significant role to predict future human health as a preventive measure-the assessment of these twin data namely blood groups and secretor status may help to reduce the morbidity and mortality of mankind and interestingly can contribute a great lot if they are added as part of one s identification data. REFERENCES 1. Maton, Anthea, Hopkins J, Mclaughlin CW, Johnson S, Warner MQ, Davidlahart, Jill D. Wright: Human biology and health Adamo PJD, Kelly GS. Metabolic and immunologic consequences of ABH secretor status. Alternative Medicine Review 2001 Aug; 6(4): Bancroft J, Gamble M. Theory and practice of histological techniques. 6th ed. 4. Godkar PB, Godkar DB. Textbook of medical laboratory technology 2003; Roy MN, Chatterjee JB. Some observations on the secretions of blood group substances in the body fluids of man. J Indian Medical Association 1965 Oct;45(8): Grundbacher FJ. Immunoglobulins, secretor status and the incidence of rheumatic fever and rheumatic heart disease. Human Hered 1972;22(4): Zhiburt BB, Chepel AI, Serebrianaia NB. The Lewis antigen system as a market of IHD risk. The Arkh 1997;69(1): Kaufmann F, Frette C, Pham QT, et al. Association of blood group related antigens to FevI, wheezing and asthma. Am J RespirCrit Care Med 1996 Jan;153(1): Dickey W, Collins JS, Watson RG, et al. Secretor status and Helicobacter pylori infection are independent risk factors for gastroduodenal disease. Gut 1993 Mar;34(3): Peter WH, Gohler W. ABH-secretion and lewis red cell groups in diabetic and normal subjects from Ethiopia. Exp Clin Endocrinoal 1986 Nov;88(1): Shinebaum R. ABO blood group and secretor status in the spondyloarthropathies. FEMS Microbiol Immunol 1989 Jun; 1(6-7): Arneberg P, Kornstad L, Nordbo H, Gjermo P. Less dental caries among secretors than among nonsecretors of blood group substance. Scand J Dent Res 1976 Nov;84(6): Thom SM, Blackwell CC, Mac callum CJ, Weir DM, Brettle RP, Kinane DF, Wray D. Nonsecretion of blood group antigens and susceptibility to infection by Candida species. FEMS Microbiol Immunol 1989 Jun;1(6-7): ABOUT THE AUTHOR S Jeelani Reader, Department of Oral Medicine and Radiology, Indira Gandhi Institute of Dental Science, Pondicherry, India, jeelani1978@rediffmail.com GS Asokan Associate Professor, Department of Oral Medicine and Radiology Tagore Dental College and Hospital, Ratinamangalam, Vandalur Post Tamil Nadu, India M Manjunath Professor and Head, Department of Oral Medicine and Radiology Vokaligara Sangha Dental College and Hospital, Bengaluru Karnataka, India Josy Jacob Professor, Department of Pedodontics, Karpaga Vinayaga Institute of Dental Science, Kancheepuram, Tamil Nadu, India 260