قراءة كتاب Contagious Abortion of Cows
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inoculating these cultures into other animals. Neither of the germs obtained in culture could therefore be regarded as the causative agent in the disease.
In 1895 Bang and Stribolt undertook the investigation of this disease, and their results are now generally regarded as the most important of all the contributions to the study of this subject. A cow showing all the symptoms of impending abortion was purchased and slaughtered. The unopened uterus was removed to the laboratory where it was opened with special precautions to avoid all contamination. An abundant, grayish yellow, odorless exudate was found between the ovum and the inner wall of the uterus. Upon standing this exudate separated into two layers, a reddish yellow serum above, and a grayish yellow partly solid layer below. In microscopic preparations of this exudate, stained with Loeffler's methylene blue, numerous very small bacilli were found, apparently in pure culture, some of them lying free, but large numbers of them crowded together inside cells. These latter appeared at first to be micrococci, but more careful examination proved them to be really short rods. Bang and Stribolt were able to cultivate this organism in tubes of a gelatin-agar-serum medium, the germ developing only in a particular zone beginning about 5 mm. beneath the surface of the medium and extending downward 10 to 15 mm. After considerable work with cultures, they concluded that the bacillus is neither an aerobe nor an anaerobe, in the ordinary sense, but exhibits a very peculiar behavior in respect to oxygen, requiring for its development a partial pressure of oxygen somewhat less than that present in the atmosphere. They were unable to obtain growth of the germ in the presence of the ordinary atmosphere, nor in the absence of oxygen (Pyrogallol method). Curiously enough, by placing their tube cultures in an atmosphere of pure oxygen, they obtained cultures developing in two zones, one near the top and the other near the bottom of the tube, indicating that there are two optima in the oxygen requirement of the organism. This very interesting character of the organism received great attention at their hands, but nevertheless Bang points out that typical development such as he has pictured was not always obtained, a number of factors seeming to cause variation in the position and extent of the developmental zones in these tube cultures. By exhausting the air above the medium in the tube, the growth was made to extend to the surface. In this way they were able to obtain growth of the bacillus on plates, but they did not work out a reliable plate method, preferring to employ the dilution tube cultures for separation in all their work. Bang and Stribolt subsequently examined pieces of placenta from a large number of cases of contagious abortion, and found the bacillus microscopically in practically all cases. Sometimes they were abundant, in other instances very scarce. Most of this material was badly contaminated, yet, from that sent in during the colder season they successfully isolated the bacillus in pure culture in a majority of the cases. In three fetuses the bacillus was found in the intestinal contents in pure culture; in one fetus it was isolated from the blood. Two cows with mummified fetus in utero were examined post mortem. These fetuses had been dead 9 months and 5 months respectively but the surrounding exudate still contained the abortion bacillus and pure cultures of it were obtained from each case. Uterine exudate kept in the refrigerator still contained living abortion bacilli after seven months.
Having found the same bacillus microscopically in a series of cases of abortion, and having obtained it in pure culture from a number of them, it now remained for Bang and Stribolt to produce the disease by inoculation of these cultures into healthy animals. Four pregnant cows were obtained without knowledge of their previous