SJMLS Volume 7 Issues 3 September 2022

seen in 14(35.9%) of the isolates. Resistance to The gradual increase in antimicrobial resistance antibiotics is on the rise and babies are in children is a major challenge in the field of particularly at increased risk because of lower hosp i t a l ep idemio logy and med ica l immunity. The study recommends the need for microbiology. This study was conducted to the development of alternative therapy. investigate the susceptibility pattern of commonly used antimicrobial agents against


Introduction of Specialist Hospital Sokoto. A structured
An estimated 30 million women living in malaria interviewer-administered questionnaire was used endemic areas of Africa become pregnant each year. to obtain some socio-demographic characteristic Pregnant women are particularly vulnerable to and blood samples were collected and examine for malaria because pregnancy reduces immunity to malaria parasite, some haematological parameters malaria. Malaria infection increases susceptibility to (HCT, HGB, WBC, RBC and platelet count) were severe anaemia and increased the risk of death measured using the HA6000 auto-haematology among pregnant women. Malaria in pregnancy analyzer (Perlong Medical Equipment Company). results in adverse pregnancy outcomes, such as Ascorbic acid levels were assayed by a standard spontaneous abortion, neonatal death, and low birth chemical method and pantothenic acid levels was weight. Chronic anaemia, due to malaria may also analyzed using the ID-VIT pantothenic acid affect a child's growth and intellectual development ELISA kit. Malaria parasitaemia and parasite load (WHO, 2006). It is estimated that at least 1 million was confirmed by the examination of a Giemsa-people die of malaria particularly pregnant women stained thin blood film. Data generated was and children less than 5 years of age, more than 80% analyzed using SPSS 22.0 statistical package. A p-of the deaths worldwide occur in sub-Saharan Africa (WHO Malaria Report, 2005). value 0.05 was considered significant in all statistical comparisons. The HCT, HGB RBC and Ascorbic acid is a natural water-soluble vitamin Platelet counts of the parasitized subjects were (Vitamin C). Ascorbic acid is a potent reducing significantly lower compared with the controls and antioxidant agent that functions in fighting (p<0.05). Also, the ascorbic acid level of bacterial infections, in detoxifying reactions, and parasitized subjects was significantly lower in the formation of collagen in fibrous tissue, compared to that of controls. There was a strong teeth, bones, connective tissue, skin, and correlation between ascorbic acid and platelet capillaries. Found in citrus and other fruits, and count (r= 0.413 and p= 0.00). The mean parasite in vegetables. Vitamin C cannot be produced or count among the parasitized subjects was 5.93 ± 3 stored by humans and must be obtained in the 2.72 × 10 /ìL. Findings of this study has shown diet (Hiten and Paula, 2011). Deficiency of that malaria in pregnancy causes decrease in Vitamin C may affect immune system as well as vitamin C and some of the haematological causing anaemia which are involved in In Sokoto, there had been a number of researches resistance to malaria. on malaria (Elueze et al., 1990;Jiya, 2001) but there is paucity of published information on full Vitamin B5 is also known as pantothenic acid, or blood count, ascorbic acid and pantothenic acid Pantothenate. It is a water-soluble B-complex among pregnant women in the area. The aim of vitamin (Horvath and Vecsei, 2009). It was this study is to analyse the ascorbic acid, discovered in 1931 by chemist Roger J. Williams pantothenic acid and full blood count of (1893-1988) during his studies on the vitamin B plasmodium parasitized pregnant women in complex (Kumar et al., 2007) daily occurrence. Sokoto city is a major commerce center in leather crafts and Pregnant women are particularly vulnerable to agricultural products. As at 2006, the state has a malaria, which is a major cause of prenatal population of 3.6 million (NPC/FGN, 2006). mortality, low birth weight and maternal anaemia. It accounts for 40% of public health expenditure, 30 -Study Population 50% of in-patient admissions and up to 50 % of out- The study population for this study includes 60 patient visits in areas with high malaria transmission malaria-infected pregnant women (subject) and (WHO, 2005b). In Nigeria, malaria is endemic and 30 age-matched healthy pregnant women stable, being a major cause of morbidity and without plasmodium infection, which were mortality, resulting in 25% infant and 30% monitored as controls. Both subjects and childhood mortality (FMH, 2005a). Tragically, the controls ware recruited in the Antenatal Clinic health status of children under the age of five and Specialist Hospital, Sokoto, Sokoto Northpregnant women has remained a major barrier to Western Nigeria. Nigeria's development. Among pregnant women, malaria is responsible for more than one in 10 deaths Study Subjects/ Selection and accounts for considerable proportion of low-Inclusion and Exclusion Criteria birth-weight babies born to these mothers. These Women who meet the following inclusion babies born with low birth weight are usually at criteria were recruited in the study; pregnant higher risk of dying from infant and childhood women parasitized with plasmodium attending illnesses (RBM, 2005).

Nigeria, women who gave written informed
Informed Consent consent in their clinic and agreed to be included Written informed consent was obtained from all in the study. The following were excluded from the study participants (subjects and controls). participating as subjects in the study; nonpregnant women parasitized with plasmodium, Sample Collection healthy pregnant women that are not parasitized Whole blood was collected via venipuncture, with plasmodium and plasmodium parasitized using BD vacutainer system into K EDTA 3 pregnant women who did not offer an informed anticoagulated and plain tube under strict aseptic consent to be included in the study.
techniques. The EDTA anticoagulated blood sample was used to analyze complete blood Study Design count while sample from the plain tubes was The research was a case-control study to assess allowed to clot. The clotted blood sample was the level of some ascorbic acid and pantothenic centrifuged at 3000 rpm for ten minutes on a acid along with complete blood count bench-top centrifuge. The serum obtain was parameters of 60 Plasmodium parasitized transferred into sterile plastic tube and stored pregnant women and 30 age and gender-matched immediately until ready to be analyzed. These healthy non-parasitized pregnant women were samples were tested in the Pathology Laboratory monitored as controls visiting the Antenatal of Usmanu Danfodiyo University Teaching Clinic Specialist Hospital, Sokoto. Blood sample Hospital (UDUTH) Nigeria. The following were collected (from both subjects and controls) laboratory investigations were carried out on 3 and tested for complete blood count, Vitamin B5 K EDTA anticoagulated blood (FBC) and serum and Vitamin C.

Sample Size Determination Analytical Method
The sample size was determined using the Diagnosis of Malaria standard formula for calculation of minimum Giemsa-stained blood film was examined for the sample size: presence of malaria parasites. The slides were 2 2 (n = z pq/d ) examined under the microscope using 40× and n = minimum sample size 100× (oil immersion) objectives (Cheesbrough, z = standard normal deviation and 2005). The full blood count was carried out using probability.
the HA6000 Auto Haematology Analyzer p = prevalence of value to be estimated from (Perlong Medical Equipment Company, China). previous studies.
The analyser determines haematological q = Proportion of failure (= 1 -p) parameters which include: Red Blood Cell d = precision, tolerance limit, the minimum Count, Haemoglobin (HGB) Haematocrit is 0.05.
hours, the growth of lactobacillus plantarum is measured turbidimetrically at 610 nm-630 nm were analyzed. The mean parasite count among 3 using the Neoeldex 4700 ELISA-reader the parasitized subjects was 5.93 ± 2.72 × 10 /ìL. (Monobind Inc., USA). Serum vitamin C Table 1 presents the socio-demographic (ascorbic acid) was assayed using chemical characteristics of the Malaria parasitized method by Natelson (1971). Ascorbic acid is pregnant subjects and controls. A proportional o x i d i z e d b y c o p p e r I I i o n t o f o r m comparison of, those that use mosquito net and dehydroascorbic acid, which reacts with acidic those that are on medication showed statistically 2, 4-dinitrophenylhydrazine to form a red bissignificant difference (p<0.05). Age, gravidity, h y d r a z o n e w h i c h i s m e a s u r e d education status, fruit intake, occupation, spectrophotometrically at 520nm. ethnicity and residence show no statistical difference (p>0.05). The mean parasite count Statistical Analysis among the parasitized subjects was 5.93 ± 2.72 × Data obtained was entered into a statistical 3 10 /ìL. Table 2 indicates that RBC, HGB, HCT package (such as SPSS version 22) on a and Platelet levels were significantly lower computer to define the nature of the distribution among malaria parasitized pregnant subjects as of data for each group. Statistical differences of compared to controls (p<0.05). However, there data were analyzed using series of statistical was no statistically significant difference in the analysis such as mean, standard deviation, Chi WBC of malaria parasitized pregnant subjects -square, student's t-test, ANOVA depending on and that of the controls (p>0.05). Table 3 the nature (categorical or continuous) and revealed that ascorbic acid levels was distribution of data (normal or non-normal).
significantly lower among malaria parasitized Pearson's correlation was used to determine the relationship between sets of data. Probability pregnant women subjects as compared to (p 0.05) was used to determine the level of controls (p<0.05). There were no statistically significant for all statistical analysis. significant differences in the pantothenic acid levels between parasitized subjects and the non-Ethical Consideration parasitized controls (p>0.05). Table 4 presents Ethical approval for this study will be obtained the correlation between the pattern of full blood from the Ethical Committee of Specialist count and some biochemical parameters. Only Hospital Sokoto. platelet count showed a significant positive correlation with ascorbic acid levels (p<0.05)

Results
and there was no significant correlation between The result of 60 malaria parasitized pregnant platelet count and pantothenic acid level, HCT, women (subjects) recruited from the Antenatal HGB, WBC and RBC counts. Similarly, here Unit of Specialist Hospital Sokoto,and 30 was no significant correlation between ascorbic apparently healthy pregnant women (controls) acid and pantothenic acid levels (p>0.05).   Table 2 shows the difference in haematological parameters of the subjects and control, there is a statistically significant decrease in HCT, HGB and Platelet of the subject compared to the controls. However, the WBC count is within the normal reference range. (pantothenic acid), *=statistically significant. Table 3 Shows the difference in ascorbic acid and pantothenic acid of the subjects and controls. The ascorbic acid level was significantly lower among the subjects compared to the controls. There was no significant difference in pantothenic acid levels between subjects and controls.   Table 4.4 shows correlation between ascorbic acid, pantothenic acid and haematological parameters. There is a positive correlation between platelet count and ascorbic acid where as there was no correlation between all the other parameters.

Discussion
In this study, the level of education was found to Malaria is a major public health problem in sub-have influence on prevention of malaria in Sahara Africa including Nigeria, where it pregnancy. Majority of the study subjects has no accounts for more cases of infection and death formal education (41%), this is followed by those than other countries in the world. The aim of this who attained secondary level education (25%) study was to investigate the effect of malaria on while women educated to primary and tertiary some haematological and biochemical level each constituted 16% of the subjects. This parameters in pregnant women attending finding is consistent with previous reports antenatal clinic in Specialist Hospital Sokoto. A (Hamidu et al., 2003;Ali et al., 2005) in Karachi, total of 90 pregnant women participated in this India and Maiduguri Nigeria respectively. This is study. The subjects were aged 15-40 years. Our suggestive that the level of education can play a finding is consistent with a previous report role in preventing malaria infection. (Sa'idu et al, 2015) in Sokoto which indicated that, young maternal age contributed to the This study indicated that the HGB, HCT, RBC seroprevalence of malaria parasitaemia among and Platelet count was significantly lower pregnant women.
The mean values of WBC count in both infected We observed that younger women in the age and non-infected women were within normal group 21-25 years constituted a significant reference range. For continued survival and number of the subjects (45%) compared to older reproduction, plasmodium parasites need to age group 36-40 (6%). This finding is consistent infect the red blood cells of their human host. with a previous report of Panti and Colleagues Consequently, changes in the red blood cell (2010) who reported that majority of the indices are some of the commonest observations asymptomatic malaria positive pregnant women seen in malaria. Anaemia, which is a fall in (84%) were aged between 20 and 34 years.
haemoglobin level below the normal range for Uneke and Colleagues (2007) in Southern age, gender, race, or pregnancy status, is the most Nigeria also reported that individuals of age frequent outward manifestation of such changes. group 20-24 have the highest prevalence of Malaria is the most common cause of severe maternal malaria (52%) while the least was anaemia in endemic areas (Abdalla 2004). Our recorded among those > 40 years. Susceptibility findings compared favorably well with the to plasmodium parasitaemia has been linked to findings of other workers in Nigeria (Onwukeme the level of antibodies to placental sequestrated and Uguru, 1990;Akingbola et al., 2006) which parasites (Elliot et al., 2005). This may be indicated that infected patients tended to have attributed to the fact that majority of the younger significantly lower platelets, haemoglobin and women are likely to be primigravidae and are red blood cell count. The lower HCT HGB and expected to have higher malaria parasitaemia.
RBC may reflect anaemia which is mainly due to This also supports the existing knowledge that mechanical destruction of parasitized red blood high prevalence at lower age is due to the cells as well as splenic clearance of parasitized existence of low natural immunity to infectious and defected red cells. The reduced platelet count diseases including malaria at that age.
in malaria is said to be due to platelet activation, splenic pooling and decrease platelet life span This study has also found that use of insecticide- (Beale et al., 1972;Abdulla, 2004). In normal treated mosquito net has great influence in pregnancy, the physiological change in preventing malaria, and is significant (p<0.05) as haemoglobin concentration (Hb) and platelet 75% of the study subjects happens not to be using count during pregnancy are well known mosquito net whereas 86.7% of the controls are phenomena (Yip, 2000). Anaemia due to Malaria users of mosquito net. This confirms the report of in pregnancy is an important public health World Health Organization (WHO 2011) and concern in developing countries (more Ntonifor and Veyufambom, 2016) on the use of pronounced in primigravidae than in insecticide treated mosquito nets as a means to multigravidae). However, anaemia due to reduce the lethal impact of malaria. malaria can be more severe in pregnant women (McCrae et al., 2011). Anaemia due to malaria significantly higher in comparison to noninfection results from the obligatory destruction malaria infected patients (Bakhubaira, 2013). of parasitized erythrocytes, the accelerated Previous report indicated that parasitized destruction of normal erythrocytes, and variable patients have higher WBCs count compared with dyserythropoiesis. Anaemia due to malaria community controls (Ladhani, 2002). The most infection can be assessed by the measurement of common complication during malaria infection packed cell volume (haematocrit) and the is thrombocytopenia (Erhart et al.,2004).

measurement of Haemoglobin concentration
Persons with platelet counts? <? 150,000/ìL (James et al., 2008). Pregnancy outcome is were 12-15 times more likely to have malaria influenced by many factors some of which infection than persons with platelet include culture, environment, socioeconomic counts? >? 150,000/Ìl (Erhart et al.,2004). status and access to medical care. The Leukocytes play a vital role in the defense haematological profile of pregnant women also against malaria. Leukocyte changes in malaria has an impact on pregnancy and the outcome of are variable and depend on many factors such as the pregnancy (Burrows et al., 2009). The most acuteness of infection, parasitaemia, disease common haematological indices are the severity, state of the host immunity to malaria, indicators of haemoglobin concentration. Low and concurrent infections (Abdalla, 2004). haemoglobin in the blood (anaemia) is widely Commonly, majority of patients with acute identified as a haematologic abnormality and it is uncomplicated P. falciparum malaria usually associated with adverse pregnancy outcome have their mean total leukocyte count (TLC) (Miltchel et al., 2006). Anaemia in malaria is within the normal range (Haroon et al.,2013). believed to occur due to haemolysis of However, in some cases, a mild leucopenia may parasitized and non-parasitized RBCs, occur, especially in non-immune adults or in peripheral removal/sequestration of RBCs, and cases of complicated malaria (Reiley and Barett, ineffective erythropoiesis (due to high 1971). The mean Total leucocyte count (TLC) in circulating tissue necrotic factor (TNFá)) parasitaemic patients in this study was 5.93 ± 3 (Akhtar et al.,2012). In malaria endemic areas, 2.72 × 10 /ìL, which is in agreement with results the prevalence and severity of anaemia are from prior studies (Tailor et al.,2008). usually determined by a number of interacting Nevertheless, despite the fact that the mean TLC factors. These include, among others, the in parasitaemic patients was normal, an increase parasite species, level of parasitaemia, age of in the level of parasitaemia in this study was host, host genetic factors (e.g., coexisting RBC associated with a decrease in the number of polymorphisms like haemoglobinopathies, leukocytes. These findings are similar to those G6PD), and nonmalarial causes of anaemia (e.g., from a retrospective study done in a malaria infections, malnutrition) (Abdalla, 2004). As endemic Indian province involving 334 patients observed elsewhere (Chandra and Chandra, with acute malaria caused by P. vivax, P. 2013), the mean red blood cell indices (Hb, falciparum, or dual infection in which a MCV, MCH, MCHC, and RDW) of patients with significant decrease in the mean TLC in the acute uncomplicated malaria in this study were parasitaemia group was also observed (Chandra normal. This could probably have been because and Chandra, 2013). uncomplicated malaria is associated with milder biochemical changes, for example, a lower In addition, according to previous studies, production of cytokines, less endothelial cell leukopenia does not appear to be parasite specific as activation, milder changes in the coagulation exemplified in a study on patients with P. vivax profile, less sequestration, and less haemolysis infection in Panama (Reiley and Barrett, 1971), in as opposed to complicated/severe malaria. Turkey (P. vivax) (Koltas et al.,2007), and in another study on 404 American service men from Vietnam Malaria infected patients tended to have with P. vivax, or P. vivax, and P. falciparum dual significantly lower platelets, WBCs, infection (Goldstein, 1968) where a leukopenia was lymphocytes, eosinophils, RBCs and Hb level, observed in majority of patients that had malaria while monocyte and neutrophil counts were caused by several different malaria parasite species.
Overall, the changes seen in the total lymphocyte parasitemic patients in this study (P = 0.72). A count in malaria parasitaemic patients are few other studies that looked at eosinophils in usually attributed either to an increase or a malaria found low levels (eosinopenia) in decrease in the differential white blood cell majority of patients (Davis et al.,1991), although (WBC) lines. In consideration of lymphocytes the significance of these findings was unknown. for instance, there have been varying reports However, follow-up of these patients' days or from different studies on whether the differential weeks after treatment surprisingly revealed a lymphocyte count remains normal, increased, or marked elevation in the eosinophil count (Davis decreased in an acute malaria infection. Pre-et al.,1991), a feature that the researchers 1970s literature had indicated that lymphocyte attributed to the rebound eosinophilic response count remains normal during an acute malaria that resulted from of an enhanced T helper-2 infection (Abdalla, 2004). However, most recent response that occurred during the malaria literature shows that lymphopenia, which is recovery period. sometimes profound but transient or temporary, is a common finding in acute malaria in Thrombocytopenia is a major complication of nonimmune adults (Richards et al., 1998) as well malaria (Khan et al., 2012), the magnitude of as in children found in malaria endemic areas which is dependent on the parasite species or (Maina et al., 2010). Monocytosis has been one disease severity. In light of the above, P. vivax of the most consistent observations reported malaria infection and severe malaria have been from prior studies done on the haematological associated with a more heightened and severe changes that characterize malaria (Abdalla, thrombocytopenia than P. falciparum infection 1988). These findings are in agreement with our and uncomplicated malaria. In this study, study, where a significant (p = 0.01) mild although the mean platelet count in parasitaemic 3 monocytosis was observed in parasitaemic patients (172.43 ±80.41 × 10 /ìL) was normal, it patients (10.89 ±6.23%) compared to the nonwas significantly (p = 0.00) lower than that of the parasitaemic patients (8.98 ±5.02%).
non-parasitaemic group (217.82 ±95.96 × 3 10 /ìL). This only reiterates the fact that acute The mean neutrophil count was normal for both uncomplicated malaria is not associated with a parasitaemic (46.28 ±18.30 %) and nonmarked reduction in platelets, as compared to parasitaemic patients (42.87 ±15.77%) in this severe malaria. In an attempt to compensate for study. These findings are similar to those from two the low absolute platelet count, the bone marrow studies: one involving 400 cases in a malaria increases the formation of megakaryocytes, endemic region of India, in which about 85% of which usually escape from the bone marrow as the patients had normal neutrophil counts (Akhtar mega platelets during an acute malaria infection. et al.,2012) and another in Singapore where Evidence to support this hypothesis comes from majority of the adults with acute uncomplicated a study by Kreil et al. (2000), that found a marked malaria had normal neutrophil counts (Kueh and elevation in the level of thrombopoietin, a key Yeo, 1982). In contrast, though, some earlier platelet growth factor in patients with malaria. studies had reported neutropenia (Dale and Wolff, Because of an increase in the number of mega 1973) or neutrophilia (Abdalla, 2004) among platelets, the mean platelet volume is increased malaria cases, especially in the paediatric patients during an acute malaria infection (Maina et al., (Maina et al., 2010). The mechanism of 2010). In contrast, the mean platelet volume neutropenia in malaria has been postulated to (MPV) of parasitaemic patients in this study was involve increased margination and sequestration normal. These findings may suggest that of neutrophils (Dale and Wolff, 1973) as a result of uncomplicated malaria is associated with mild or the increased expression of cell adhesion nonsignificant changes in the platelet profile. molecules (ICAM-1 and VCAM-1) that occurs in The pathogenesis of thrombocytopenia is malaria (Clark et al.,2006). thought to involve a constellation of processes, some of which include splenic pooling of The eosinophil count was not significantly platelets, antibody (IgG) mediated platelet different between the parasitemic and nondestruction, adenosine diphosphate (ADP) release following the hemolysis of parasitized the risk of urinary tract infection in pregnancy RBCs, dysmegakaryopoiesis, platelet (Ochoa-Brust et al.,2007). The recommended aggregation and activation, parasite invasion of daily intake of Vitamin C among pregnancy in the platelets, platelet phagocytosis, platelet adhesion EU (European Food Safety Authority (EFSA), to erythrocytes, and oxidative stress (Lacerda et 2017) and United States (Panel on Dietary al.,2011). The relatively diverse causative Antioxidants and Related Compounds,2000) is pathophysiological mechanisms could probably 105? mg and 85? mg respectively. explain why changes in platelet homeostasis are more prominent than in other blood cell lines.
In this study, the pantothenic acid levels among Nevertheless, thrombocytopenia in malaria is malaria parasitized pregnant women subjects observed to improve with disease resolution, and showed no significant difference when compared a normal platelet count is usually reported within with the controls. There seems a paucity of data on 7 days after the initiation of antimalarial pantothenic acid levels among malaria parasitized treatment (Moulin et al., 2003). pregnant women subjects. Pantothenic acid is a component of coenzyme A (CoA) and The results of this study indicated a significant phosphopantetheine, both of which are involved in decrease in the ascorbic acid (vitamin C) level of fatty acid metabolism. It is widely distributed in malaria parasitized pregnant women when foods and the deficiency is only seen in individuals compared with control (p<0.05). This is in who are fed with synthetic diets (Fry et al., 1976) or consonance with the finding of Boudová et al.
in those fed an antagonist (Hodges et al., 1958). Our (2014) who reported that the mean Vitamin C finding is consistent with a previous report which level were lower in malaria positive pregnant indicated that the average pantothenate blood level than malaria negative pregnant women. Malaria of the pregnant women was lower than that of noninfection is associated with increase production pregnant controls (Song et al., 1985). Our finding is of ROS by phagocytes, this change may play a at variance with a previous report (Srinivasan and vital role in host defense against malaria and it Belavady, 1976) which indicated that pregnant could also render host tissue such as erythrocytes subjects had blood levels of the pantothenic acid more susceptible to oxidative damage (Vasquez comparable to those of the non-pregnant controls. et al., 2003).
Plasmodium parasitaemia may be a factor responsible in the reduced pantothenic acid among Ascorbic acid is potent water-soluble antioxidant our cohort of parasitized pregnant women. Previous and the biological system utilizes it in report indicated that it is important for dieticians, scavenging/neutralizing an array of Reactive nutritionists, physicians, and other healthcare Oxygen Species (ROS) which were produced at providers to be able to offer accurate and evidencevery high level because of increased activity of based advice on supplement use in pregnancy. NADPH oxidase of immune cells (Une and Gupta, Nutritional supplementation may be a safe and cost-2013). The decrease in vitamin C is therefore effective way to reduce risk of outcomes among attributed to increase in its consumption as pregnant women. antioxidant vitamin in clearing the Reactive Oxygen Species (ROS). Vitamin? C deficiency is In this study, we observed that there is no prevalent particularly among low-income correlation between the haematological populations, pregnant women, pregnant smokers parameters and ascorbic acid and pantothenic and pregnant women with type 1 diabetes acid except for a positive correlation between (Madruga de Oliveira et al., 2004;Schleicher et al., Platelet count and ascorbic acid level. There was 2009;Juhl et al., 2016). Previous report no similar study in relation to these findings. This recommends regular vitamin C supplementation observation might be due to the fact that vitamin during pregnancy and that it reduces C functions to maintain healthy blood vessels hospitalization (Hans and Edward, 2010).
which protect small vessels from damage and Vitamin? C supplementation has been shown to also helps in wound healing. It also prevents reduce the risk of premature rupture of the hardening of the vessels and blood clot (Iqbal et membranes (Stuart et al.,2005) as well as reduce al., 2004) amongst others.