Proteolitik Olmayan, Avirulan Bacillus Antracis Mutantlarının İzolasyonu veKobaylarda Mukayeseli Bağışıklık Denemesi ile Bu Mutantlardan Canlı Aşı Hazırlanması

Year 1969, Volume: 3 Issue: 7-8, 23 - 54, 01.12.1969

Şükran Durukan Nabi Emre

Abstract

Bu çalışmada Sterne suşu 'nun proteolitik olmayan mutantla riyle hazırlanan anthrax aşıları ve orijinal Sterne aşılannın mukayeseli bağışıklık, zararsızlık deneyleri ve aşının maliyetini etkileyen faktörler üzerinde durulmuştur . 1 - 20 cm. mesafeden 35 watt'lık ultraviole ışınlarına 45 dakika maruz bırakmakla Bac. anthracis'in Sterne suşundan (34F2 ) avirulan, proteolitik olmayan suşlar izole edildi. Proteolitik olmayan mutant suşların sütlü agarlardan besleyici agarlara pasajları yapıldı ve aerop ve % 20 co, atmosferi altında 3 er gün üretildi. Bunlardan bir çoğu proteolitik olmayan karakterini kaybetti, fakat bir mutant suş (29/la) bu şekilde yapılan 45 pasajda proteolitik olmayan karakterini muhafaza etti. 2 - Aynı metodla virulan, bir suştan da 2 proteolitik olmayan mutant suş izole edildi. Bu proteolitik olmayan mutant suşların 500 sporu kobayların deri altına injeksion edildiğinde virulan bulundular. 3 - 29 /la ve Sterne suşları, kazein digest vasatarında besleyici agarlara nazaran daha iyi sporlandı. Kazein digest vasatlarında 3 gün üretilen 29 /la mutant suşunun glyserinli süspansionu Sterne suşunun glyserinli süspansionuna nazaran % 25 daha fazla canlı spor ihtiva etmekteydi. Kazein digest vasatlarında 5 gün üretilen 29 /la suşunun süspansionları Sterne suşuna nazaran % 100- 150 daha fazla canlı spora sahipti. 4 - 29/la ve Sterne suşlarının kobaylarda yapılan mukayeseli bağışıklık testlerinde 29 /la mutant suşu, Stern e suşuna nazaran 2 - 5 misli daha etkili bulundu. S - Bu suşların 2x10 8 ·Ve 1x10 8 miktarlarındaki sporları deri altı ve adale içi kobaylara inakule edildiği zaman 29 /la mutant suşu Sterne suşundan daha emin bulundu. 6 - Ml. lerinde 8, 6, 4 ve 2 milyon spor bulunan 3 çeşit [a - % 40 glyserin, % 60 fizyolojik tuzlu su ve % 0,1 sapanin b - % 30 aluminium hydroksit, % 70 fizyolojik tuzlu su; c - Mineral yağlı Saf White Nedle Oil- B) ] aşı 29 /la mutant suşu ile hazırlandı. Keza Sterne aşıları da aynı miktar spor ihtiva edecek şekilde hazırlandı. Bu aşılardan her birinin O,S ml. i ile deri altı verilmek suretiyle 6 şar koyun aşılandı ve aşılamadan 7 ve 15 gün sonra eprüve edildi. Mineral yağlı aşı hariç her iki suştan da hazırlanan bütün aşılar çok iyi bağışıklık verdiler. 7 - Ml. inde 10 milyon spor bulunan her bir aşıdan 1- 3 ml. ile koyun ve keçiler deri altı ve adale içi inokule edildiği zaman Sterne aşısı 29 /la mutant suşundan hazırlanan aşılara nazaran daha çok reaksion verdi.

Keywords

Sterne suşları, Sterne suşu, Mineral yağlı Saf White Nedle

The isolation of avirulent nonpreteolytic mutant of Basillus antracis and the immunity test in guinea pigs and preparation of live vaccines from this mutant

Abstract

This study has been done comparatively on the immunity and safety test on the vaccines which were prepared by a nonproteolytic mutant of Sterne's strain and the vaccines of original Sterne's and the factors of financial effect of production. 1 - Some avirulent, nonproteolytic mutants were isolated from Sterne's strain (34F2) of Bacillus anthracis by exposure by irradiation for 45 minutes at 20 cm. from the filament of a 3S W Ultraviolet lamp· These nonproteolytic mutants were madde subcultures from the milk agar to nutrient agar and were incubated aerobic and 20 % carbon dioxide atmosphere for 3 days. Many of them lost their nonproteolytic character, but a mutant strain (29/la) was stable its nonproteolytic character in about 4S subculteres by the same method. 2 - 2 nonproteolytic mutants were isolated from virulent strain by the same method. These nonproteolytic mutants were found virulent, when they were tested in guinea- pigs for virulence by subcutaneous injection of SOO spores. 3 - 29/la and Sterne's strain both sporulated better on casein digest medium than nutrient agar. The glycerinized suspensions of 29 /la mutant strain contained about 2S % mo re viable spores than the glycerinized suspensions of Sterne's strain, when they were incubated on casein digest medium for 3 days. The suspensions of 29 /la contained ab out 100- 150 % more viable spores than the suspensions of Sterne's strain, when they were incubated on casein digest medium for 5 days. 4 - Comparison of the protection of guinea- pigs induced by the vaccines of the 2 strain showed that the 29/la mutant strain was from 2- to S times more effective than the Sterne's strain. 5 - 29/la mutant strain has been found safer than Sterne's strain, when guinea- pigs were inoculated intramusculary and subcu taneously with about 2 x 10 8 and 1 x 10 8 spores of each of these strains. 6 - Three kinds of vaccnies [a - 40 %" glycerine, 60 % physiological saline, and 0,1 % saponin b - 30 % aluminium hyddroxide, 70 % physiological saline; c - mineral oil (White Needle Oil- B)] which included 8, 6, 4, and 2 million spores in ı ml. of each, were prepared by 29/la mutant strain. Sterne's vaccines were prepared also in the same amount of spores. 6 sheep of a group were vaccinated subcutaneously with O,S ml. of each of these vaccines and challenged on 7 and ıs days after vaccination. All the so vaccines which were prepared by both strains, except the vaccines of c- with mineral oil, gave very good immunity. 7 - When sheep and goats were inoculated intramusculary and subcutaneosly with 1-3 ml. of each of these vaccines which contained 10 million spores per ml., Sterne's vaccine gave more reactions than the vaccines which were prepared from 29/la mutant strain.

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