Screening for hepatitis B virus ( HBV ) , hepatitis C virus ( HCV ) and syphilis infections among asymptomatic students of a private university in Western Delta , Nigeria

This study was aimed at screening for non-treponemal antibodies, anti-hepatitis C and hepatitis B surface antigen markers among asymptomatic students of Western Delta University, Oghara, Nigeria. Venous blood was collected from 318 students of 26 years average age who did not show any visible signs or symptoms of any of the three infections. Blood samples were processed and screened for hepatitis C, hepatitis B and syphilis by rapid test methods. Blood samples were collected and processed in the Microbiology and Biotechnology Laboratory of Western Delta University, Nigeria between June 2013 and October, 2013. Anti-hepatitis C assay was done by a one step hepatitis C virus test strip, hepatitis B assay was done by a one step hepatitis B surface antigen test strip and non-treponemal antibodies was done by syphilis ultra rapid test strip. Ninety three (29.3%), 36 (11.3%), 6 (1.9%), 3 (0.9%) and 0.0% number of students belonged to 21-25 (24 years average), 15-20 (19 years average), 26-30 (27 years average), 36-40 (40 years average), 41-45 (44 years average) and 31-35 (30 years average) age groups respectively. Nine (5.1%) male students out of the 117 (55.7%) screened in the 21-25 age bracket tested positive to hepatitis B (produced antibodies against HBsAg) while 0.3 (1.7%) female students of the same age group produced antibodies to Treponema palladium antigen (tested positive to syphilis). Seropositivity to HBsAg was also recorded by 3 (8.3%) male students out of 36 (11.3%) screened in the 26-30 years age group. On the whole, 12 (3.8%) male and 3 (0.9%) female students tested positive to hepatitis B and syphilis, respectively. Hepatitis due to HBV and syphilis infection are prevalent within the university under study and indeed, Oghara town though in apparent low occurrence. The relevant health authorities should ensure a constant sero-epidemiological surveillance as well as institute counseling and vaccination programmes.


INTRODUCTION
Most cases of viral hepatitis in humans are caused by four viruses.These include hepatitis B virus, hepatitis C virus, hepatitis A virus and hepatitis D virus or delta hepatitis (CDC, 1991).There are, however, distinct differences in these viral agents in terms of their epidemiologic, immunologic and clinical characteristics (Robinson, 1990).For drug users, hepatitis viruses B and C are the agents of concern because the major risk of infection is by blood-borne transmission especially through contaminated needles (CDC, 1991).
Hepatitis B virus (HBV) is a double stranded DNA virus which is transmittable by three main mechanisms which include: (1) percutaneous (2) sexual contact and (3) mother-to-child (CDC, 1991).Hepatitis B surface antigen (HBsAg) has been isolated in all body fluids such as blood products, saliva, semen and vaginal fluids (CDC, 1991).After the initial exposure, the incubation period averages 60-90 days and sometimes, 6 weeks to 6 months (CDC, 1991).Infected persons may be sources of infection to others before the onset of symptoms which include loss of appetite, vague abdominal discomfort, weight loss, fever, nausea, vomiting and diarrhoea.Others are dark tea-coloured urine, muscle aches and skin rashes prior to the development of clinical jaundice (CDC, 1990).
Hepatitis C virus (HCV) was first isolated in 1988 (CDC, 1990).Previous studies have confirmed that the hepatitis C virus is the viral agent that causes non-A, non-B hepatitis (Benenson, 1990) and it is associated with both acute and chronic liver disease.HCV, like HBV is a blood-borne pathogen which is transmitted by direct contact with infected blood products, contaminated needles or syringes, sexually as well as through vaginal secretions, semen or saliva (Benenson, 1990).Symptoms of HCV hepatitis include vague abdominal discomfort, anorexia, nausea, vomiting and jaundice with elevated liver enzymes (CDC, 1990).
Syphilis is a systemic infection that has been known since the 15 th century.It is caused by the spirochaete-Treponema pallidium which is spread by contact with infectious lesions during sexual intercourse (Hook and Marra, 1992).About 12 million people worldwide are infected with syphilis each year and most of them live in developing countries (WHO, 2001).
HCV, HBV and syphilis infection are very common in HIV-infected persons (Soriano et al., 2008;Laurent et al., 2003).Patients with HCV and HBV co-infection with HIV have an increased risk of progression of HCV and HBVrelated liver diseases (such as chronic hepatitis, cirrhosis and hepatocellular carcinoma) when compared with HCV mono-infected patients (Konopnicki et al., 2005;Chen et al., 2006;Omland et al., 2009;Ananthakrishnan et al., 2010).
There is paucity of documented or published literature (information) on the seroprevalence of HBsAg, anti-HCV and non-treponemal antibody (syphilis) in Nigeria and perhaps Africa.The purpose of this work was therefore to study the seroprevalence of HBsAg, anti-HCV and nontreponemal antibody (syphilis) markers among asymptomatic students of a private University in Western Delta, Nigeria with the under mentioned objectives: (a).
Analyse the sex and age distribution of Western Delta University students recruited for the study, (b).Determine the presence of HBsAg, Anti-HCV and syphilis antibody markers among apparently healthy students recruited for the study.

Informed consent
The researchers obtained a written informed consent from students who were mainly adults and able to make independent decision.Students were given an informed consent questionnaire which they filled and signed.Oral informed consent was also given by students engaged in the study.

Sampling
Four millilitres (4ml) of venous blood was collected by vein puncture with sterile 5ml needles and syringes from a total of 318 students aged 15-44(have age 26yrs) and made up of 108 (34.0%) males and 210 (66.0%) females of Western Delta University, Oghara, Nigeria.Subjects were grouped into nine departments which included Geology, Accounting, Mass communication, Computer Science, Microbiology and Biotechnology, Business Administration, Biochemistry, Economics and Political science.Subjects were also grouped into 15-20, 21-25, 26-30, 31-35, 36-40 and 41-45 age brackets with the average age for each age group recorded.Venous blood samples were dispensed into appropriately labelled ethylene-diamine tetra acetic anti-coagulated blood containers and properly mixed with the anticoagulant.Blood samples were allowed to stand on working bench for about 30minutes to allow for proper plasma separation by force of gravity.

One step hepatitis B surface antigen test strip (rapid test procedure)
A rapid one step test for the qualitative detection of HBsAg in the plasma samples was done according to the method initially prescribed by Blumberg et al. (1971).The kit for this rapid test is commercially available.

One step hepatitis C virus test strip rapid test procedure
A rapid one step test for the qualitative detection of antibodies to hepatitis C virus in the plasma samples was carried out according to the method described by Choo et al. (1989).The kit for this rapid test is commercially available

Syphilis ultra rapid test strip procedure
The syphilis ultra-rapid test strip assay was carried out according to the procedure described by Johnson and Taylor-Robinson (1994).The kit for this rapid test is commercially available.*Corresponding author.E-mail: fesdotaj@yahoo.com.
Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution License 4.0 International License (5.1%) male students tested positive for hepatitis B surface antigen.In this same age group, the male students recorded 0 (0.0%) and 0 (0.0%) antibody in vitro response to HCV and syphilis assays, respectively.The female students in the  21-25 age bracket recorded 0 (0.0%), 0 (0.0%) and 3 (1.7%)seropositivity to HBsAg, anti-HCV and nontreponemal antibody, respectively.A total of 36 (11.3%) students were in the 26-30 age group of which 3 (8.3%),0 (0.0%) and 0 (0.0%) male students showed reactivity to HBsAg, anti-HCV and syphilis antibody, respectively.The female students in this bracket recorded 0 (0.0%) each, reactions to HBsAg, anti-HCV and syphilis antibody, respectively.The 31-35 age group had no student recorded in it.In the 36-40 bracket, out of 6 (1.9%) students, both the male and female students recorded 0(0.0%) sero-reactions to HBsAg, anti-HCV and non-treponemal antibody respectively.Similarly and lastly, all the 3 (0.9%) students in the 41-45 age group tested negative to hepatitis B, hepatitis C and syphilis.In all, 12 (3.8%)male and 3 (0.9%) female students tested positive to hepatitis B and syphilis respectively.No student tested positive to hepatitis C.

DISCUSSION
Screening tests do not mean the prevalence of infection but the prevalence of the markers which are important from epidemiological perspective (Silvio and Johnson, 2011).Hence this work sought to unravel the seroprevalence markers of HBsAg, anti-HCV and nontreponemal antibodies (syphilis) among healthy (asymptomatic) students of Western Delta University, Oghara.Apart from presentation or observance of symptoms, the risk factors involved in the disease caused by the above agents justify screening for the diseases or their markers in populations (Shiell and La, 2001;Castelnuovo et al., 2006).
In this study, out of the 12 (3.8%)HBsAg seroprevalence rate recorded, 9 (5.1%) and 3 (8.3%)prevalence rates occurred among male students in the 21-25 and 26-30 age groups, respectively.The seroprevalence rate of hepatitis B in this study is low and this result is consistent with prevalence rates of 3.0 and 3.5% as reported in similar studies by some authors (Carey et al., 2007;Moukoko et al., 2014).Conversely, the result obtained is at variance with hepatitis B prevalence rates of 16.0, 18.0, 19.7, 22.4 and 49.1% as published by previous authors in similar studies (Tabibian et al., 2008;Chang et al., 1993;Almeida and Pedroso, 2004;De-Souza et al., 2004;Kulik, 1999).No female students recorded positive reactivity to HBV antigen among all the age groups.Conversely 3 (0.9%) syphilis seroprevalence rate found in this research occurred in female students all of whom belonged to the 21-25 age bracket.
The occurrence of HBsAg and syphilis in both male and female students may be incidental and may have no direct connection with gender although this was not tested statistically.This opinion is supported by Zhao et al. (2011) who did not establish any gender association with the disease or microbial agents described in this study.Besides, it has been established that variations in prevalence rates may be due to different patterns of injection used by drug users and different geographical locations (Cad-Saude et al., 2008).Vellinga et al. (1999) however reported that risk factors for hepatitis B infection among people with intellectual disability include male gender, old age and geographical location.
Syphilis disease has been associated with migration and population.Seroprevalence rate of 3 (0.9%) for syphilis was recorded in this study.This low prevalence may be due to the fact that the students concerned (or most of the students) are not migrants from place to place.According to Macpherson and Gushulak (2008), syphilis has been associated with migration and the international movement of disease.The 0.9% syphilis seroprevalence is inconsistent with 1.1, 1.1, 1.5, 3.35, 3.3 and 19.6% prevalence rates published by some authors (Guimaraes et al., 2007;Cad-Saude et al., 2008;Silvio and Johnson, 2008;Takada et al., 2003;Carey et al., 2007;Zhao et al., 2011).
Findings in this study showed no case of co-infection between any of the three microbial agents.This is in contrast to some reports which established a significant association between HCV and syphilis infection (Hwang et al., 2000;Adjei et al., 2008;Wu et al., 2010).A similar situation was seen in the 3 (8.3%)HBsAg seropositive cases in the 26-30 age group.The only 3 (1.7%)seropositive reactivity for syphilis which was recorded by all females in the 21-25 age bracket had no record of coinfection with any of the other agents.
The non-association of HCV infection with syphilis infection as recorded in this study is however consistent with some reports which stated that a majority of studies suggest that HCV is not associated with syphilis in general but with genital ulcers inherent in syphilis infection (Marx et al., 2003).The students who were seropositive for HBV antigen and syphilis antibodies did not show physical signs of the disease.It is possible sooner than later, they will.This deduction is corroborated by Konopnicki et al. (2005), Chen et al. (2006), Omland et al. (2009) and Ananthakrishnan et al. (2010) who reported that patients with HCV and HBV coinfection have an increased risk of progression of HCV and HBV related liver disease (chronic hepatitis, cirrhosis and hepatocelular carcinoma) when compared with mono-infected patients.Although students used for this investigation were not screened for anti-retroviral (HIV) antibodies, the obtained results somewhat suggest that students and indeed subjects in and around Oghara town may be almost free from the diseases implicated.This is against the background of documented reports that state that HCV, HBV and syphilis infection are very common in HIV-infected persons (Soriano et al., 2008;Laurent et al., 2003).

Conclusion
Overall occurrence or prevalence rates of 3.8% for HBsAg and 0.9% for syphilis in this study indicate that hepatitis due to HBV and syphilis infection are being harboured by students of Western Delta University and perhaps, residents within and around Oghara town though in apparent low occurrence.Health authorities of Ethiope West LGA and indeed, Delta State should therefore start off or ensure constant seroepidemiological surveillance in order to monitor the prevalence of hepatitis, syphilis and other infections within the study area.Counseling and vaccination programs would also help to control their transmission.

Table 1 .
Sex and age distribution of Western Delta University students recruited for the study.

Table 2 .
Sero-prevalence of hepatitis C (HCV) antibody, hepatitis B surface antigen (HBsAg) and non treponemal antibody (syphilis) among students recruited for study.