Diagnostic Study of Venereal Disease Research Lab (VDRL), Malaria Parasite (MP) and Blood Grouping in Human Blood Donors

Our current study was conducted on the Prevalence of VDRL, MP and Blood grouping of Human blood donors in University of Swat. The aim and objective of our current study is to determine the prevalence of VDRL and MP and distribution of blood groups among males, district Swat. For this purpose, A totals of 65 blood donors’ were screened for the above disease, out of which, 24 collected from Al-Fajar Foundation, Gulkada and 41 from Frontier Foundation, Mingora. The results shows that the donors were non-reactive for VDRL and MP whereas the ABO blood groups distribution among males, were 32.30%, was group A, 35.30% was group B, 10.7% AB and 21.50% were O. The prevalence of A+ blood group were 32.3%, B+ blood group were 32.3%, O+ blood group were 21.5%, AB+ blood group were 9.23%, AB- blood group were 1.53% and the O- and A- blood groups were not found among male blood donors. This study concludes that, there are no reactive or positive cases of VDRL and MP among male blood donors, so that’s why there is no prevalence of VDRL and MP among male blood donors of the District Swat, and the A+ (32.3%) and B+ (32.3%) blood groups are very common in the swat and the blood group O- and A- are very rare. Disease Research Laboratory; FTA-ABS: Fluorescent ANC: Antenatal Care;


Introduction
Venereal Disease Research Laboratory (VDRL) test is a nontreponemal test, used for the screening of syphilis. VDRL test still the most used test throughout the world. And still remains unchallenged. CSF VDRL test is very specific for neurosyphilis [1].
There are three basic methods used for syphilis screening such as dark field microscopy, notreponemal and treponemal to identify spirochete. The more sensitive nontreponemal tests are VDRL and RPR use for early screening; while specific treponemal test is Fluorescent Treponemal Antibody Absorption (FTA-ABS) use to confirm diagnosis [2]. Syphilis infection caused by Treponema pallidum belongs to spirochete group of bacteria; also assist the transmission of HIV [3,4]. According to World Health Organization about 12 million endure the syphilis throughout the world in every year. The oral and genital mucus are more sensitive to Treponema pallidum bacteria [5]. Syphilis infection can be transfer from mother to child [6]. Mother-to-child transmission (MICT) of syphilis during pregnancy can cause a serious fetal outcomes in the second or third trimester such as early fetal death, stillbirth, neonatal death, preterm birth, low birth weight and congenital infections in infants [7]. Syphilis is the second most common infection causing of stillbirth worldwide [8]. Preventing MTCT of syphilis through expanded early testing in antenatal care (ANC) and immediate treatment with a single injection of benzathin penicillin [9], is highly cost-effective [10,11].
Human blood and blood products are used for the prevention and treatment of life threatening-disease and also reduced transfusion transmitted infections (TTIs) such as syphilis infections, HIV, HBV, and HCV [12]. The transfusion of unscreened or screened blood and blood products are the major source of TTIs in developing countries like India [13]. It has been found that genetically determined ABO blood group antigen which is present in blood may block binding of possible TTIs causative organism to polysaccharides on cell but non-secretor are lacking in their antigen and are at risk to variety of infections TTIs [14]. Blood group means the entire blood group system contain red blood cells (RBC) antigens that are controlled by specific genes. Karl Landsteiner has been credited for the discovery of ABO blood group system in 1900 [15]. Discovery of ABO blood group system extend the way for discoveries in the field of immune haematology, blood transfusion among humans [16]. The ABO blood group is divided into four blood types (A, B, AB, and O) on the basis of presence or absence of A and B surface antigens. ABO blood group is major because of the certainty that A and B are strongly antigenic and anti A and anti B is naturally develop antibodies present in the serum of individuals require the analogous antigen.
These antibodies have ability to produce intravascular hemolytic in case of incompatible transfusion [17]. The Rh (Rhesus) system is the second most important blood group system in human blood transfusion with presently 50 antigens.
The most important antigen is the D antigen, because it produces can produce IgG anti-D antibodies [18]. Rh blood group was discovered by Landsteiner and Wiener in 1941 [19]. Rh antigens are largely immunogenic. D-negative persons produce anti-D if they encounter the D antigen through transfusion or pregnancy and causes hemolytic transfusion reaction of fetus and newborn.
For this reason, the Rh status normally determined in blood donors, transfusion recipients and in mothers [20]. Recently studied that, ABO group not only for blood transfusion but also associated in various infectious disease transmission such as malaria, salivary gland tumors, colorectal cancer, carcinoma of stomach, thyroid disorder, ovarian tumors and small cell lung cancer [21]. Malaria is the most deadly parasitic infection in the worldwide. Malaria can transmit by blood transfusion, from mother to fetus and by using of infected needle. Malaria is generally transmitted by female Anopheles mosquito acting as a vector to disperse the genus of Plasmodium [22]. Plasmodium is transmitted to human by mosquito biting. This Plasmodium either enter into the RBCs or hepatocytes through the blood streams [23]. There are five species of Plasmodium known to cause malaria, such as; P. malariae, P. vivax, P. falciparum, P. ovale and P. knowlesi.P. vivax and P. falciparum species are mostly common in Pakistan [24].
Human Red Blood Cells (RBCs) or erythrocytes are the major sites of these parasites infection. The infected person suffers from the high fever, chills, nausea and anemia. Now is malaria known as the tropic disease [25]. Malaria transmission period for P.vivax peaking from June to September and again in April to June, when relapses on infections acquired the previous season are observed, while the transmission period for P.falciparum is between August and September in Pakistan [26]. The lifecycle of Plasmodium is very complex. Malaria caused in human by the transmission of plasmodium species through the mosquito female Anopheles. And these Plasmodium species transmits into the vertebrates [27].
Mostly female mosquito takes blood meals, because she needs supplementary blood meals to produce eggs. When a female mosquito bites it transmits sporozoites from its saliva into the human blood stream. Studies have shown that salivary duct of mosquito are blocked by these parasites, as results of which mosquito require more effort to achieve food [28]. Small amount of sporozoite engulf by macrophages and other by lymphatic system where they become destroyed. In the skin it is suppose that about 10% of the parasite remains which mature into infectious merozoites [29,30]. After entering of sporozoites into the liver produce [35]. The merozoites change into the gametocytes (male and female gametes) within the bone marrow. The gametocytes are mostly present in the patient's blood, so mosquito takes it during blood meals. Gametocytes of P. falciparum take 7-15 days and 1-3 days in other species, to absorb in blood [36,37]. Gametocytes mature in gametes and fertilization occur in midgut of the mosquito and gametes change into the motile zygote known as ookinetes.
Apoptosis occurs in 50% of the unknown reason. And the rest of ookinetes run away from the midgut through the periphery of the midgut membrane, and penetrates the liver and here it divides a few times to produce sporozoites. These sporozoites then go to salivary gland and during blood meals it inject into the blood of the host [38].

Aim and Objectives
The aim and objective of this study is to determine the prevalence of VDRL and MP and distribution of blood groups among males in swat (Figure 1).

Data Collection
The study was conducted from July to December 2020 at University of Swat, Town Ship Kanju, Swat, Khyber Pakhtunkhwa, Pakistan. The blood was collected in blood bags from the all age of male students from different departments by the Informed Oral Consent. Total of 65 blood samples were collected in which, 24 blood samples were collected by the Al-Fajar Foundation team, Gulkada, Saidu Sharif Swat at November 2020 while 41 were collected by Frontier Foundation, Mingora, at July 2020. The blood samples were taken based on the body weight (50<kg), it means that only those males had donated the blood whose had more than 50kg weight and also based on the previous disease.

Data Analysis
The data was collected through the informed oral consent and then analyzed to determine the prevalence of VDRL and MP tests results among males and also determined the distribution of blood groups. So it was recorded that there were simple percentage of blood groups and non-reactive VDRL and MP among males.
There were 0% prevalence of VDRL and MP negative among males, all of 100% negative (Figure 2).

ABO Blood Groups among Males
were positive. In our study no reactive cases of VDRL were there.
A study was reported by [43][44][45][46] in Karachi, Pakistan. They studied on 800 pregnant women. The prevalence of syphilis among women in Ibrahim Hyderi was 1.5 % higher than in other sites 0.5%. In our study there were no VDRL reactive cases among the males in swat.

Conclusion
This study concludes that there is no prevalence of VDRL and MP among male blood donors. All of 65 blood samples are VDRL and MP negative. And out of 65 male blood donors 14 (21.53%) were O+, 21 (32.3%) were A+, 21 (32.3%) were B+, 2 (3.0%) were B-, 1 (1.5%) were AB-and 6 (9.2%) were AB+ whereas A-and Oblood groups were not found. So, it concludes that the blood groups A-and O-are very rare in the Swat and A+ and B+ blood groups has very high frequency as compare to previous studies, it shows that the A+ and B+ blood group are very commonest blood groups in the District Swat.