Determination of Adsorption Capacity of Alum Hydroxide {Al (OH) 3} Gel for Streptococcus equi sub specie equi and Streptococcus dysgalacltae sub species equisimillis

Abstract

Citation: Manzoor S, Rahman S, Ashraf M, Khan FM, Syed ZH (2017) Determination of Adsorption Capacity of Alum Hydroxide {Al (OH) 3} Gel for Streptococcus equi sub specie equi and Streptococcus dysgalacltae sub species equisimillis. J Antivir Antiretrovir 9: 006-009. doi:10.4172/1948-5964.1000155 Copyright: © 2017 Manzoor S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Determination of Adsorption Capacity of Alum Hydroxide {Al (OH)3 AbstractThe present study was conducted to determine the adsorption capacity of Alum Hydroxide gel for Streptococcus equi and Streptococcus equisimillis in combination. One ml of streptococcal inoculum containing live count of Streptococcus equi @ 2 × 10 9 /ml and Streptococcus equisimillis @ 2 × 10 9 /ml inoculated in each of six Eppendorf tubes containing 0.2 mg, 0.4 mg, 0.6 mg, 0.8 mg, 1.0 mg and 2.0 mgs of autoclaved Aluminium Hydroxide in the form of gel, mixed and centrifuged at 1600 rpm for 15 minutes produced supernatant, which upon streaking and incubation on seven nutrient agar plates including supernatant from 7 th control negative Eppendorf tube containing sterilized normal saline produced different number of colonies after adsorbing streptococcal cells depending upon the concentration of Al(OH) 3 in gel. 50 colonies were counted from supernatant recovered from Eppendorf containing 0.2 mg of Aluminium Hydroxide, 25 colonies from supernatant over 0.4 mg, 15 colonies from supernatant over 0.6 mg, 10 colonies from supernatant over 0.8 mg and no colony was obtained from supernatants over 1.0 mg and 2.0 mgs of Aluminium Hydroxide while 100 colonies were recovered from control negative ependorf containing only normal saline without Aluminium Hydroxide. It is concluded that Aluminium Hydroxide as gel should be used as adjuvant @ 1 mg per ml of streptococcal inoculum in a streptococcal vaccine. Journal of Antivirals & Antiretrovirals Preparation of formalin-inactivated Streptococcus equi and Streptococcus equisimillis antigenSelected colonies of S. equi and S. equisimillis isolate were inoculated in two 500 ml flasks separately containing Modified Todd-Hewitt broth supplemented with 5% horse serum. Both the flasks were kept on orbital shakers at 60 rpm for 48 h. After that formalin (0.2%) was added in both flasks to kill the S. equi and S. equisimillis isolate. These bacterial isolates were kept stable for 24 hours for proper action of Formalin. The killed organisms from both flasks were harvested by centrifugation at 6000 xg for 1 h at 4°C. Two washings with sterile PBS (pH 7.2) were done. The pellets from both flaks thus obtained were re-suspended in PBS. The immunogenic concentration of S. equi and S. equisimillis as declared by Manzoor et al. was adjusted as 2 × 10 9 per ml and 2 × 10 9 per ml respectively by spectrophotometer. These preparations were stored at 4°C until utiliz...