EFEK ANTIVIRAL RIBAVIRIN DALAM PERTUMBUHAN DAN PERKEMBANGAN EKSPLAN BAWANG PUTIH CV. LUMBU HIJAU, CV. LUMBU KUNING DAN CV. TAWANGMANGU

Tanaman bawang putih (Allium sativum L) termasuk dalam genus Allium yang diperbanyak secara vegetatifmelalui umbi. Virus merupakan salah satu penyakit penting yang perlu dipecahkan pada pembiakan vegetatif ini.Teknik inkonvensional kultur jaringan yang dikombinasikan dengan kemoterapi dapat membantu menghilangkanpenyakit virus. Penelitian ini bertujuan untuk melihat pengaruh dari beberapa konsentrasi antiviral ribavirin dimedia MS terhadap pertumbuhan dan perkembangan shoot tip Bawang putih cv Lumbu Hijau, cv. Lumbu Kuning,cv. Tawangmangu. Percobaan dilakukan di laboratorium kultur jaringan, Balai Penelitian Tanaman Sayur(Balitsa), pada bulan Mei hingga Juli 2015. Sasaran penelitian adalah untuk menghasilkan tanaman bebas virusdengan menggunakan teknik kultur jaringan yang dikombinasikan dengan kemoterapi. Varibel yang diamatiadalah pertumbuhan dan perkembangan planlet bawang putih. Hasil dari penelitian (1) Kontaminasi kulturumumnya disebabkan oleh bakteri dan jamur dengan persentase 10 % sampai dengan 30%. (2) Penambahanantiviral ribavirin, semakin tinggi konsentrasi persentase tumbuh dan berkembang semakin rendah untuk ketigakultivar (3) Pengamatan secara visual penambahan antiviral ribavirin dan kultivar tidak berpengaruh pada jumlahtunas, rata-rata dari satu eksplan tumbuh satu tunas untuk ketiga kultivar (4). Penambahan antiviral ribavirin dankultivar tidak mempengaruhi pertumbuuhan daun, akar ketiga kultivar (5).Hasil pengujian virus dengan teknikDAS ELISA persentase kultur yang terinfeksi 54.55% sampai dengan 100 %.Kata kunci: bawang putih (Allium sativum L); antiviral ribavirin; kultivarABSTRAKThe garlic (Allium sativum L) belonging to the genus Allium, propagated in vegetative through bulb. Inthe plants propagated by vegetative technique, virus is an important disease to be solved. The tissue culturetechniques in combination with chemotheraphy could eliminate virus diseases. The experiment carried out in thelaboratory tissue culture, Balai Penelitian Tanaman Sayur (Balitsa) on May untill July 2015. The experiment aimsto observe the effect of several antiviral ribavirin concentration in MS medium on growth and development shoottip cv. Lumbu hijau , cv. Lumbu kuning , cv. Tawangmangu. It’s main goal is to produce virus-free plants usingtissue culture techniques combined with chemotheraphy. The variables observed were the growth and developmentof garlic plantlets. The results of the experiment are; (1). Culture contamination were generally caused by bacteriaand fungi with a percentage of 10% to 30%. (2) In the high concentration of antiviral ribavirin gave results ondecreasing growth and development of the three garlic cultivar (3) On visual observation, cultivar and antiviralribavirin has no effect on the number of shoots, each explants were growing one shoot. (4). The added of antiviralribavirin and cultivar does not affect on growth the three garlic cultivar . (5) The results of the test virus byserological test DAS ELISA techniques the percentage of infected culture were 54.55% to 100%.Key word : Garlic (Allium sativum L); Antiviral ribavirin; cultivar

A Polymerase Chain Reaction–Based Method for the Detection of Hepatitis A Virus in Produce and Shellfish

Outbreaks of gastroenteritis that are suspected to be of viral origin are on the rise. Thus, there is a need for regulatory agencies entrusted with food safety to develop adequate techniques for the detection of viruses in foods. We have established a general procedure for the detection of hepatitis A virus (HAV) in shellfish that, with minor modifications, is also applicable to fresh produce such as cilantro. Total RNA was isolated from shellfish or cilantro, followed by isolation of poly(A)-containing RNA. Because HAV genomic RNA contains a poly(A) tail, the isolation of poly(A)-containing RNA also enriches HAV genomic RNA. Reverse transcription was used to convert the RNA to cDNA, and then amplification was carried out by polymerase chain reaction (PCR). Reamplification with internal primers was used to improve the quality and the quantity of amplified DNA, allowing for post-PCR analysis such as sequence identification of the viral strain. With this procedure, multiple samples could be analyzed in four working days by a single trained individual. The nominal sensitivity of detection of the procedure was 0.15 TCID50 (50% tissue culture infective dose) per 0.62 g of tissue with a test virus. The direct RNA isolation protocol avoided pitfalls associated with whole-virus purification procedures by replacing virus precipitation steps involving polyethylene glycol and Procipitate with phenol extraction. The method is straightforward and reliable. We successfully used this procedure to detect naturally occurring HAV in clams involved in a gastroenteritis outbreak, as well as in cilantro artificially contaminated with a test virus.

Activity of an Alcohol-Based Hand Gel Against Human Adeno-, Rhino-, and Rotaviruses Using the Fingerpad Method

Objective:To assess the activity against three non-enveloped viruses (an adeno-, a rhino- and a rotavirus) of a gel containing 60% ethanol, using experimentally contaminated thumb- and fingerpads of 12 panelists, as per standard procedure E-1838-96 of the American Society of Testing and Materials.Design:Each digit received 10 μL of the test virus suspension. The inocuLum from the thumbs was eluted immediately with 990 μL of Earle's balanced salt solution (EBSS) to assess the amount of virus on each digit (0-minute control). The inoculum on the fingers was allowed to dry (20-25 minutes), and virus was eluted from two fingerpads to determine the loss in virus infectivity upon drying (baseline titer). Then the dried inoculum on randomly selected fingers was exposed to 1 mL of the test product or standard hard water (200-ppm calcium carbonate) for 20 seconds. The virus remaining was eluted with 1 mL of EBSS, titrated to determine the amounts eliminated, and compared to the baseline titer.Results:Each digit received at least 104 plaque-forming units of virus in 10 μL The amounts of adeno-, rhino-, and rotaviruses surviving the drying were 30%, 75%, and 42%, respectively. The product reduced the infectivity titers of the three viruses by 3 to >4 log10when compared to a reduction of ≤ 1 log10for the hard-water rinse.Conclusion:The level of virus reduction by gel was statistically significantly higher than that seen with the water control. Evidence for such activity against non-enveloped viruses supports further investigation of the benefits of this product.

FURTHER CHARACTERIZATION OF THE ADENOVIRUS ERYTHROCYTE RECEPTOR-MODIFYING FACTOR

Agglutinability of human erythrocytes for 3 hemagglutinating adenoviruses was markedly reduced by pretreatment of red cells with a factor present in tissue cultures which had been infected with adenovirus types 1, 2,4, or 15. The factor responsible for erythrocyte receptor modification was non-dialyzable and unaffected by the action of ribonuclease, desoxyribonuclease, trypsin, chymotrypsin, or ether. The factor was smaller, more thermostable, and separable from the infectious virus. Erythrocyte receptor modification was found to be a function of time and temperature. Titers of erythrocyte receptor-modifying activity were not diminished by successive exposures to fresh erythrocytes. Erythrocytes treated with erythrocyte receptor-modifying factor suspensions failed to significantly adsorb test virus hemagglutinin. Inhibition of erythrocyte receptor modifying-activity of the adenovirus suspensions by rabbit antiserum was type-specific.

STUDIES OF MOUSE POLYOMA VIRUS INFECTION

Three procedures have been compared for usefulness in titration and detection of polyoma virus: production of cytopathic effect (CPE) in mouse embryo tissue culture, production of HI antibody after inoculation into weanling mice (MAP test), and production of tumors in suckling hamsters during a 3 to 5 week observation period. The tissue culture and mouse antibody production tests were generally comparable in sensitivity, reproducibility, and time required to obtain results. Titration by tumor production in suckling hamsters was not suitable for quantitation because of marked variation in susceptibility among animals. Virus was detected in tissues of normal mice from spontaneously infected colonies by either production of CPE in mouse embryo tissue culture or by the MAP test; virus was found in organs of 15 (58 per cent) of 26 mice with antibody, and 2 (8 per cent) of 24 mice without antibody.