DEVELOPMENT OF ENZYME-LINKAGE IMMUNOSORBENT ASSAY AGAINST TYPE B OF CLOSTRIDIUM BOTULINUM: A PRELIMINARY STUDY

Clostridium botulinum neurotoxin (BoNTs) is one of the causes of economic loss in the livestockindustry. This economic loss would be as a direct result when animals poisoned by BoNTs or indirectlywhen the livestock products are contaminated by BoNTs, which end up with the products are banned byauthority. Therefore a routine surveillance of BoNTs in the farm and in livestock product processingindustry is urgently needed. One of the most relatively quick and accurate methods to perform a routinedetection of the presence of BoNTs is enzyme-linkage immunosorbant assay (ELISA). In this article wedescribe the results of the development of ELISA, using polyclonal antibodies against BoNTs-Bproduced locally. Antibodies were generated from six Balb/c mice with standard immunologicalmethods. Mice were immunized three times for a period of 8 weeks with a commercial type BClostridium botulinum toxoid at a dose of 100 ng per mouse per injection. The resulting antibody waspurified by a combination of ammonium sulfate precipitation 50% (w/v) technique and a protein Acolumn method. The results of this preliminary study indicated that the developed ELISA methodcapable of detecting type B Clostridium botulinum toxin up to 1.0 ng/ml

Penghambatan Proliferasi Limfosit Mencit Balb/c oleh Ekstrak Testis Sapi Bali: Peran TGF-β

Testis extract (ET) of several animals such as rats, mice, boars, and bulls, has been reported able to suppress lymphocyte proliferation. In order to elucidate if ET of bali cattle has the suppressive activity, the present study was conducted. ET was isolated aseptically from the testis of bali cattle slaughtered at a slaughter house in Mataram, Lombok. After decapsulating, the testis were minced, homogenized (1 g tissue per 3 ml phosphate buffered saline, PBS, pH 7.2). The ET (40, 20, and 10µg/ml) was added on to lymphocytes isolated from Balb/c spleen, then cultured for 72 hours in RPMI media containing 10% foetal bovine serum in 96 well plates with the concentration of 250,000 cells per well, with- or without concanavalin A (10 µg/ml), or IL-2 (0.2 ng/ml). After 72 h of culture, cell proliferation was examined by colorimetric assay. The results showed that bali cattle ET were able to suppress the proliferation of lymphocytes in vitro through the process of apoptosis. Western blot analysis revealed that bali cattle ET contain TGF-β1. The suppression effects of the ET were eliminated by anti-TGF-β1 antibody. It can be concluded that ET of bali cattle expressed TGF-β which inhibited lymphocyte proliferation through an apoptotic cell mechanism. The actual roles of TGF-β in the bali cattle testis need to be elucidated further

UTILIZATION OF POLYCLONAL ANTIBODIES PRODUCED IN LOCAL HORSES (EQUUS CABALLUS) AS A RESOURCE FOR DEVELOPMENT OF ELISA CONJUGATE TO DETECT HEPATITIS B VIRUS (HBV) SURFACE ANTIGENS

The aim of this study was to utilize the antibody produced using Indonesia local horses (Equuscaballus) to make the conjugate of ELISA kit for detection of hepatitis B virus (HBV) surface antigen(HBsAg). The polyclonal antibodies were isolated and purified from local horses immunized repeatedlyusing isolated and purified HBsAg from Indonesia. The antibodies were conjugated with horseradishperoxidase by a modified method of Nakane and Kawaoi. The conjugate activities were performed usingthe principle of ELISA test conducted by the researchers as well as by independent laboratory.Commercial conjugate for HBsAg ELISA was used as a comparison study. The results of this studyindicated that the antibody produced from local horses can be used to make conjugates that werecomparable to commercial HBsAg ELISA kit