Prevalence and genotype distribution of hepatitis B virus among migrant workers in Lombok Island, Indonesia

To examine the potential risk of hepatitis B virus (HBV) spread in Indonesia by migrant workers, based on the molecular characteristics of HBV strains. Methods: Sera collected from migrant workers traveling to their destination countries (pre-migrant workers) and those returning to Indonesia (post-migrant workers) were screened for HBsAg by ELISA, followed by HBV DNA detection by PCR and (sub) genotype/subtype determination according to surface region and whole genome sequencing. Results: Of 87 pre-migrant workers, 15 (17.24%) were HBsAgpositive, whereas 15 (12.10%) of 124 post-migrant workers were HBsAg seropositive. HBV genotype analysis based on the S region showed that HBV-B3/adw2 was predominant (96.15%, 25/26) whereas 3.85% (1/26) of isolates were HBV-C3/adrq+. Whole genome sequencing of selected strains and phylogenetic tree analysis identified subgenotype B7 in three samples previously categorized as subgenotype B3 based on S region analysis, supporting a recent argument that subgenotypes B5/B7/B8/B9 could be considered as a quasi-subgenotype of B3. Conclusions: A high prevalence of HBsAg carriers was detected among migrant workers from Lombok Island, with no significant difference in prevalence between before and after returning to Indonesia. All strains were classified into genotypes common in Indonesia, and the results suggested that migrant workers are not a risk factor for HBV transmission into Indonesia.

Acute Oral Toxicity of Immunoglobulin Y (IgY) anti HIV in Mice.

The production of antibodies in chickens and the extraction of specific antibodies from egg yolk (IgY antibodies) are increasingly attracting the interest of the scientific community, as demonstrated by the significant growth of the IgY literature. The objective of the study was to evaluate the oral acute toxicity of IgY anti HIV on Mice. In acute toxicity study, mice by administering once orally graded doses of the IgY anti HIV in the ranges of 0.9375 g to 15g /kg body weight and observed for 14 days and the number of dead mice was recorded and used in the calculation of the acute toxicity value (LD50). The mice were also observed for other signs of toxicity, such as convulsion, diarrhea, cornea reflex, dyspnea, righting reflex, straub. Oral administration of IgY anti HIV at dose of 0.9375; 1.875; 3.75; 7.5; 15 g/kg body weight showed there no mortalities or evidence of toxicity effects, suggesting that the LD50 value of IgY anti HIV was more than 15000 mg/kg body weight. Throughout 14 days of the treatment no changes in behavioural pattern, clinical sign of toxicity, vital organs weight (liver, lung, heart, spleen and kidney) and body weight of mice in both control and treatment groups. Also there were no any significant alterations in the biochemical analysis of the blood serum (SGPT, SGOT, BUN and Creatinine).The overall finding of this study indicates that the oral administration of IgY anti HIV did not produce any significant toxic effect and practically non toxic in mice. Hence, the IgY anti HIV can be utilized for immunotherapy on HIV patient.

Production and characterization protein of anti HIV specific immunoglobulin Y for Immunotherapy.

In this study, an approach for large-scale production of immunoglobulin Y ( Ig Y) specific anti Human Immunodeficiency Virus (HIV) for use as a potential immunotherapy for HIV disease. Lohman laying hens were immunized intramuscularly with HIV virus that had been inactivated using formaline. The immunizations were repeated two times with dose of each 1 ml of HIV antigen (20 copy/ml) with an interval of two week. The first immunizations were HIV antigen mixed with Fruend Adjuvant Complete and subsequently mixed with Freund Adjuvant Incomplete. Egg yolk was separated from egg white and immunoglobulin Y (IgY) antibody was then purified by multiple polyethylene glycol (PEG) 6000 extraction and ammonium sulfate purification steps. Antibody response in yolk was detected by agar gel precipitation test (AGPT) and the protein pattern was detected using polyacrilamid gel electrophoresis (SDS-PAGE). Specific activity of the antibody was tested using commercial ELISA. Antibody of HIV was detected and produce a specific line of precipitation in AGPT beginning the second week after the first immunization. Analysis of results obtained with ELISA showed significant increase in the HIV-specific antibodies after two weeks from the primary immunization. Through the effect of boostering; the anti-HIV antibody levels reached a plateau at six weeks from the primary immunization and remained significantly higher till the end of observation period. SDS-PAGE revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 164; 87; 68, and 37 kDa. These results suggested that chicken IgY could be a practical strategy in large-scale production of Ig Y specific anti HIV for immunotherapy and diagnostic KIT of HIV disease.