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Bacillary White Diarrhea*
POULTRY SCIENCE VOL. IV
AUGUST-SEPTEMBER, 1925
No. 6
BACILLARY WHITE DIARRHEA* F. R. BEAtTDETTB New Jersey Agricultural Experiment Station
* Paper No. 198 of the Journal Series, New Jersey Agriculturay Experiment Station, Department of Poultry Husbandry.
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Since bacillary white diarrhea Avas described by Rettger in 1900, its presence has been determined in every part of the United States. The disease has been found in Europe and future investigation will no doubt show that it is present in every locality where poultry raising thrives. The ease with which this disease can be introduced into a new locality makes its spread all the more possible. The economic importance of bacillary white diarrhea has attracted many to its investigation and the information thus derived has been plentitful. In fact, it can be said that more is known about this disease than any other malady that affects the common fowl. In order to appreciate this fact, it might be well to scan the various contributions that have elucidated the nature of this disease. Rettger,^ in 1900, was the first to describe the organism that is responsible for white diarrhea, though at that time the disease was described as a septicaemia and its relation to what is now known as bacillary white diarrhea was probably not suspected. On this occasion his observations were confined to a brood of seventeen chicks, all but three of which became sick. Of the fourteen sick chicks eleven died and three finally recovered, though they remained stunted in growth for two months or more, after Avhich they grew rapidly again. The symptoms and post mortem findings of the disease were described quite fully. From the livers of the chicks, and in one case from the spleen, a small bacillus was isolated and described. The organism was reported as being motile, an observation which was later found to be incorrect by the Same worker.
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In order to determine the pathogenicity of this organism, four chicks ranging in age from two to four weeks were inoculated subeutaneously with 0.5 cc. of a twenty-four hour bouillon cul ture. All of the chicks became sick and two died. A third was killed after it had showed signs of recovery. From the two chicks that had died and from the third that was killed the original or ganism was isolated from the livers and in one instance from the lungs and kidneys. Mention was made of the fact that the mother hen as well as other grown chicks were apparently im mune, and it is to be noted also that in the inoculation experi ment the younger chicks died, whereas the older ones recovered. In 1901, Rettger- again reports his findings in regard to a similar disease of larger proportions occurring at Winona Lake. In a lot of forty chicks (four broods), thirty-five died while between one and four weeks old. In another brood of fifteen the disease appeared before the chicks were a week old. Later, two other broods of thirty-three chicks became affected and in less than three weeks all but ten had died. The disease next ap peared on two adjoining farms where 80 per cent of the chicks died that contracted the disease. The organisms isolated from the livers of the chicks at this time corresponded with that described previously by the same author. By means of this organism he Avas again able to cause the death of two chicks (two weeks old) by subcutaneous injection. The economic importance of the disease as manifested by its epi demic nature was noted at this time. In 1906, Graham^ reports a disease of young chicks at the Storrs Station. The disease appeared in a February hatch of 400 chicks and it is stated that nearly all of them died. A January hatch of 150 chicks and a March hatch of 500 chicks were apparently not affected. The disease was undoubtedly white diarrhea and this outbreak probably marks its introduc tion into this Station, where it was thoroughly investigated in later years. In 1908, Rettger and Harvey* report a third epidemic that occurred in Connecticut in 1907. In this outbreak one hundred and forty-six chicks were exposed to the disease and eighty-four or about sixty per cent died. The bacillus isolated resembled the organisms obtained from the tAvo previous epidemics. In
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describing the organism it is stated to be non-motile, whereas it was previously said to be motile. The cultural features of the organism are again reported and its ability slightly to acidify litmus milk is noted for the first time. The fermentation reactions of the organism are also reported, for the first time. Two guinea pigs inoculated with 1.5 cc. subcutaneously and 5.5 cc. intraperitoneally with broth cultures respectively proved refractory. It was found that a rabbit was not killed by three injections of living culture after it had been immunized with several injections of dead culture. The disease was produced for the first time by feeding the organism to two chicks ten and eight days old. Death occurred on the fourth and nineteenth day respectively. It was noted on this occasion that the serum of a chick that had been injected with killed cultures would agglutinate the homologous organism. The same reaction was also noted in connection with the serum of an immunized rabbit. The identity of this disease with what is commonly known as white diarrhea is brought out. The causative organism is placed for the first time in the colon-typhoid group. The source of infection is still unsolved, though it was brought out that transmission of the disease from chick to chick might take place through the medium of contaminated food and drinking water. Whether the egg is infected before it is laid, during incubation, or not until the chick is hatched, are yet unsolved problems. In 1908, Milks'^ describes an outbreak that came to his attention in 1907. The disease was confined to chicks under five or six weeks old. The symptoms and post mortem lesions described were undoubtedly those of white diarrhea. An organism was isolated whose cultural features and fermentive powers are identical with the organism described by Rettger. The occurrence of grayishwhite areas on the heart is noted for the first time. In 1909, Rettger* again describes the disease and calls attention to the almost constant presence of an unabsorbed yolk in chicks dead of this disease. Mention is also made of a semi-solid or cheesy plug found in the ceca. This lesion was noted by Milks in 1908. The organism is named Bacterium pullorum and methods are given for its isolation and identification. The oc-
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currence of anaerogenic strains of the organism is noted for the first time. The organism is reported as being isolated from the crop, stomach and intestines, as well as from the unabsorbed yolk. Experiments showed the organism to be particularly suscepti ble to the action of lactic acid, and it was suggested that the feeding of sour milk might have beneficial results. That the egg is suspected as a source of infection, is evidenced by the fact that over five dozen eggs were examined. However, these examina tions were negative in every ease. In 1909, Rettger and Stoneburn' record extensive observa tions made at the Storrs Station. White diarrhea had ap parently been prevalent on this plant for several seasons and it was deemed advisable to dispose of a greater part of the breed ing stock. On dressing the birds for market they were noted to be in poor condition and several showed pathological ovaries. The first hatch of the season came off in February 1909. In a lot of 250 chicks a few were reported as dying from "White diarrhea." The brooders were again filled in March, and this time serious losses were sustained. Specimens were examined, and the specific organism was isolated from the internal organs. Because a number of Rhode Island Red hens seemed to be par ticularly affected, it was decided to make a study of their eggs. The shells of twelve eggs were examined with negative results. The yolks of eighty-six incubated eggs yielded a pure culture in eight instances, five were questionable. The eggs were stated to have come from a number of Λ'arieties, but the organism was ob tained from eggs laid by Rhode Island Red hens in seven in stances and only in one egg laid by a Buff Orpington. The yolks of twelve fresh Rhode Island Red eggs were examined and the organism was recovered from one. The ovaries of twenty-three hens were next examined and twenty-one showed pathological changes. Cultures were taken from twenty of the ovaries and the organism was obtained in nine instances. Six other Rhode Island Red hens were examined and in every instance where the ova were pathological the organism was recovered. Prom these findings it was concluded that the hen is the original source of infection. The normal mortality in 280 chicks from the Station stock
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was found to be about 13 per cent. A few drops of water suspen sion of B. puUorum given to each of fif tyincubator hatched chicks caused the disease to develope in a large number and thirty-four or 68 per cent died. In another infection experiment where bouillon cultures were used, the mortality at the end of eight weeks was 47 per cent for the infected chicks and 17 per cent for the controls. The organism was recovered from about 75 per cent of the chicks that were infected. In a third experiment where the culture was supplied in the drinking water, a big dif ference Avas noted in the mortality of the infected and control lots. In 1910, Jones'* states that his attention was first called to this disease in 1910. On this occasion six one week old chicks were examined and the organism recovered. The mortality in this out break was over 90 per cent and the year previous about 1800 chicks had died apparently from the same disease. The disease appeared to have been present in this flock to some extent in 1908. Of 104 chicks produced on this plant and shipped twentyfive miles, thirty-eight remained well at the end of t ^ o weeks. In another lot of 100 chicks that were shipped, fifty-five had died. A lot of twenty eggs injected with 0.1 cc. of Bacterium pullorum failed.to hatch, although the embryo developed for about fifteen days. Another lot similarly injected with sterile bouillon produced fifteen chicks. Eggs smeared with cultures of Bacterium pullorum and then incubated produced chicks, none of which died of the disease. In 1911, Rettger and Stoneburn" published their second report of work done at the Storrs Station. Cultures of Bacterium pul lorum fed to chicks 12 hours old caused a mortality of 71 per cent, as compared with 4 per cent in the control lots. The in fected chicks Λvere stated to be at least 25 per cent lighter in weight than the uninfected chicks. The teehnic of examining fresh eggs is described wherein the whole yolk is added to a quantity of broth and incubated 72 hours. From this, tubes are streaked to determine the presence of Bacterium pullorum. It was noted that infected hens seem to be poor layers, especially during their second year. Also, that chicks hatched in winter and in early spring were comparatively free of the disease. The question of hens contracting infection as adults is still un-
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solved. The male bird as a factor in transmission is also unsolved. Testis have been examined, but with negative results. A cycle of infection is represented for the first time. It is to be recalled, however, that the way in which hens become infected is not as yet definitely established. In 1911, Gage^" reports the isolation of Bacterium pullorum from the ovaries of hens. The organism was isolated from twelve different ova of the same hen and the pathogenicity of these strains tested on baby chicks. Eighty-two chicks were used. Sixty-two were inoculated, thirty by subcutaneous injection and thirty-two were fed two drops of bouillon culture. The remaining twenty chicks were held as controls, and at the end of six weeks, all of the controls were alive and in good condition. Of the sixty-two inoculated and fed chicks, sixty died and the organism was recovered in every case. In 1911, Jones^^ showed that young chicks which recovered from the primary infection might retain the infection in their ovaries, as evidenced by examinations of their eggs and ovaries. It was also shown that ovarian infection could be induced by intravenous injection of the organism. For detecting carriers of infection, Jones suggested the use of the agglutination test. In 1911-12, Jones" reports an epidemic in adult fowls due to Bacterium pullorum. In this instance a poultryman who had no trouble from this disease incubated eggs for a neighbor who had very poor success. The eggs produced a poor hatch and some of the incubated eggs were fed to adult hens. In sixteen days after the eggs were fed losses occurred, and in all, fifty fowls died. The organism was recovered from the pericardium, liver, spleen and ovaries of the dead hens. This farm had never had white diarrhea in baby chicks, but after the feeding of the incubated eggs, the chicks produced from this stock showed the disease, indicating that ovarian infection was undoubtedly produced by feeding of incubated infected eggs. "The infection in adults is characterized by necrotic foci in the liver, spleen and pancreas and large necrotic nodules in the heart muscle. The presence of a fibrinous exudate on the capsule of the liver and spleen and on the pericardium and heart was a somewhat constant lesion." The occurrence of nodules on the heart was first reported by Milks in 1908.
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In 1911-12, Jones^^ reports on the value of the macroscopic agglutination test in detecting fowls that are harboring ovarian infection. Of the twenty-one fowls found to give: a positive reaction by this test, twenty were shown to be carriers by bacteriological examination of the ovaries. I t was stated that probably the remaining one Avould have been found to be infected had more extensive searches been made. Unheated test fluid was found to give a little better result than the heated antigen. The methods of collecting blood samples, the making of test fluid and making the test are also described. In 1912, Rettger, Kirkpatrick and Stoneburn^* report further experiments. To determine the age of susceptibility, five lots were used containing twenty-six chicks each. One lot was not infected and remained as a control and the mortality was 11.5 per cent in this lot. Of the chicks infected at 36 hours, 33.5 per cent died ; 23.33 per cent of the chicks infected at 60 hours died ; 19.25 per cent loss was sustained in those infected at 84 hours and only 7.7 per cent of the chicks died that were infected at the age of 108 hours. In a second experiment 300 chicks were used and divided equally into six lots of 50 chicks each. 16.66 per cent of the controls died ; 72.11 per cent of those infected at 24 hours died ; 88 per cent infected at 48 hours died ; 30 pei· cent of those infected at 72 hours succumbed ; 20 per cent of the lot infected at 96 hours died and the mortality in the lot infected at 120 hours was 16.0 per cent. It was concluded from these experiments that chicks are most susceptible up to forty-eight hours after hatching. Fourteen hens were placed together, seven of which were known to be carriers and seven were known not to be infected by egg examinations. The birds were allowed to associate together for about two years and at the end only four of the originally uninfected hens remained alive. About eight months after the beginning of the experiment. Bacterium pullorum was found in one of the eggs of the uninfected birds, a little later two more infected eggs were found and it was determined that all three eggs came from the same hen. Sixteen months later, an infected egg was found from one of the other hens and a week later a second egg from thé same hen was found to contain the organism. Still later, an infected egg was laid by the third hen and on post mortem examination all four hens were
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found to have pathological ovaries. The experiment showed that infection might be contracted by association with infected hens. In order to determine whether hens could contract infection from infected litter, six pens were maintained containing 8 hens each, three of the pens served as controls. Bacterium puUorum was sprinkled on the litter of three of the pens two or three times a week. Eight White Leghorn and forty Rhode Island Red pullets were used and the experiment was carried on for one year. At the end of the year, the 24 pullets serving as controls had not laid an infected egg and none of their ovaries were found to be pathological on post mortem examination. Of the twentyfour birds in infected pens, eight had produced infected eggs and a ninth bird showed pathological ovaries on post mortem. The experiment proved beyond a doubt that infection of the ovary could be contracted from infected litter. Experiments were carried on to determine the value of sour milk as a preventive. A large number of chicks were used in several trials. In every ease, the chicks which had received sour milk showed a much lower mortality than did the infected chicks which had not received sour milk. Chicks fed sour milk were found to make more rapid gains. In 1913, Bushnell and Maurer^" report on the value of milk cultures of B. bulgaricus as a preventive for this disease. In their experiments it was shown that such milk exerted a beneficial action in that when administered 40 hours before infection, the mortality was much lower than in check pens which did not receive the milk. The feeding of B. bulgaricus milk 40 hours after infection with Bacterium pullorum prevented as high a mortality as was experienced in the check pens. In 1914, Gage, Paige and Hyland" confirmed previous observations made by Rettger, that the organism was more readily isolated from incubated eggs than from fresh eggs. The technie used was essentially the same as that described by Rettger. Their examination showed that from 4 to 21 eggs must be examined, before the organism was recovered in the experimental birds used in their work. The laying period covered fom 6 to 58 days. It was shown that hens inoculated some two years previously with a culture of Bacterium pullorum intravenously resulted in an ovarian infection.
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In 1915, Rettger, Kirkpatrick and Card^" reported results that indicated no difference in the results of feeding sweet or sour milk. Milk soured with B. bulgarieus gave no advantage over that soured naturally. The beneficial results are explained on the basis that milk feeding increased growth and vitality to such an extent that the resistance of the chick was increased and hence made it more immune to infection. Milk feeding seemed to prevent mortality from all causes. It was concluded from their
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The eggs from a number of hens were examined and it was demonstrated that a negative egg record also gave negative results on autopsy, whereas all but two hens yielded cultures on autopsy that had given a positive agglutination reaction. It is explained that these two probably retained only the agglutinin from the primary infection. Polyvalent test fluid was found,to give better results than a monovalent antigen. In 1914, Rettger"^' reviews direct transmission of the disease to the offspring. Again in 1914, Rettger, Kirkpatrick and Jones^' positively proved that chicks having the infection when young and recovering from it, retained in the ovaries a localized infection in some instances. Chicks w^ere infected when young and some recovered, when tested at the age of one year a number reacted positively to the agglutination test and on autopsy these showed pathological ovaries. Of 138 chicks growing to maturity, 88 were infected when quite young and 57 were held as controls. Of the 88 infected when young, a positive agglutination test was obtained on 21, showing that over 25 per cent retained the infection localized in the ovary. One of the 57 controls gave a positive test and also showed pathological ovaries. This was probably a contact case, as it was in close proximity to some of the infected birds. The macroscopic agglutination test was found to agree with post mortem examination. Evidence was presented which seemed to show that agglutinins would appear in the serum of fowls fed a ration infected with the organism. In one instance, a iowl gave a positive serum test, but on post mortem examination the ovaries were found to be normal. However, a thoracic tumor was found and from this a pure culture of Bacterium puUorum was isolated.
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study that milk as a food was not of value because of the acids which it might contain, but upon the natural constituents of the milk itself. In 1915, Smith and TenBroeck^" present data on a comparative study of Bact. pullorum and Bact. sanguinarium. They conclude that the two organisms are different. It was shown that Bact. pullorum could be distinguished from Bact. sanguinarium because the former was able to produce gas on dextrose and mannite, whereas the latter failed in this respect. The ability of Baet. sanguinarium to produce acid on maltose also distinguished it from the white diarrhea organism. In their «studies of the agglutination affinities it was pointed out that the white diarrhea organism was closely related to the human typhoid bacillus and the fowl typhoid organism in its serum reactions.. Rabbits were killed with bacterial free filtrates of 5 and 15 day-old cultures of the white diarrhea organism. The animals died from what appeared to be a toxic action of the organism. In 1915, Gage and Paige-^ show, by chick examination, that bacillary white diarrhea is very prevalent and widespread in Massachusetts. Various breeds were tested by the agglutination method and the range of infection varied from 0 per cent to over 50 per cent in the flocks tested. It was stated that reacting fowls were found which originated in Europe. Fowls whose serum caused agglutination in a 1-100 dilution were considered as infected. Rettger, Kirkpatrick and Jones in 1915^^^ give the results of testing 14,617 fowls contained in 107 different flocks. Of this number, 13,831 were hens and 1417 or 10.24 per cent were found to be infected while only 23 or 2.9 per cent of the 786 males tested reacted. The per cent of infection in both males and females was 9.85 per cent. Only 4.3 per cent of the Leghorns were found to be infected; 12.2 per cent of the Rhode Island Reds gave a positive test ; 13.7 per cent of the Plymouth Rocks were found to be infected and the Wyandottes reacted to the extent of 23.9 per cent, while the miscellaneous breeds showed a 17.1 per cent infection. The per cent of infection was found to be greatest in large flocks. Eleven of the 23 reacting males were killed and examined for the presence of Bacterium pullorum. In the testes of two of the
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males the organism was recovered in pure culture. In three of the males the organism was isolated from the pericardial sac. In a fourth male both the testes and pericardial sac were found to be infected. The presence of agglutinins in the six males from which the organism was not recovered is explained on the basis that they merely retained the agglutinins from the infection while young. Thirteen flocks were retested and in four cases no reactors were found on second test. The per cent of infection varied in the other flocks from 0.6 per cent to 25.7 per cent, though in every case the per cent of infected birds on second test was less than was found on the first test. In 1916, Gage and Martin'^' report studies on the histopathology on the intestines of chicks dead of white diarrhea. In a lot of 63 chicks, 53 were injected with a mixture of three strains of Baet. puUorum. Of the inoculated chicks, 26 died in from 4 to 11 days and 27 recovered. The control chicks did not die and grew normally. The occurrence of death at different ages afforded a variety of material with which to study the changes in the intestine. Sections of acute cases showed a hyperemia and catarrhal exudation with desquamation of the epithelium. In some cases hemorrhage was in evidence. Mononuclear lymphocytes were found to be numerous. In more chronic cases regeneration of the epithelium had started and proliferating fibroblasts were seen. Mucus and white cells were abundant. In chronic eases a thickening of the intestine was noted. In 1916, Rettger, Hull and Sturges^* report results with feeding experiments with Bact. puUorum to rabbits, kittens, guinea pigs and white rats. Young rabbits were found to be most susceptible and death usually took place. In kittens symptoms of severe food-poisoning were noted. The possibility of infection to man from eating infected eggs improperly cooked is pointed out. The resistance of the organism in eggs subjected to frying, coddling, boiling, poaching and scrambling was also studied. It was found that boiling for four minutes did not kill the organism in all cases. Frying of eggs hard on one side also failed to render them sterile in all cases. Prying on both sides was found to be effective as was scrambling and poaching. Rettger, Kirkpatrick and Jones in 1916^^ give a progress re-
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port on the testing of fowls for carriers of white diarrhea. In all, 7799 birds were tested, 6358 of which were retested from the year preceeding but coming from infected flocks at that time. One hundred and fiftj'^-seven males M^ere tested, 96 of which were tested for the first time. All males reacted negatively. The distribution of infection among the various breeds was found to be as follows:—Rhode Island Reds, 11.9 per cent; White Leg horns, 3.4 per cent ; Wyandottes, 10.3 per cent ; Plymouth Rocks, 11.7 per cent; Orpingtons, 31.7 per cent; FaveroUes, 22.2 per cent and miscellaneous breeds 23.2 per cent. Of all breeds, 8.2 per cent were found to be infected. Of the 78 flocks tested, 25 were found to be free of infection. Four of the Leghorn flocks were found to be free on retest. Of the seven Rhode Islajid Red flocks, five showed infection on second test. In all cases, the second test showed a lower per cent infection than the first test. All three Barred Rock flocks showed reactors on second test and in 5 Wyandotte flocks the number of carriers found on second test was much lower than found on first test. The infection was reduced from 3.5 per cent to 0.0 per cent in Leghorn flocks; in Rhode Island Red flocks from 11.25 per cent to 1.8 per cent; in Barred Rocks from 22.8 per cent to 7.6 per cent ; in Wyandottes from 20.8 to 14.5 per cent ; the reduction in all flocks was from 13.16 per cent on first test to 6.37 per cent on second test. The total number of fowls tested in the two years was stated to be 21,317 with a 9.3 per cent average infection. Of the 1037 males tested, 2.1 per cent reacted. The per cent of infection in various flocks ranged from 0.0 per cent to 56.3 per cent. It was stated also that from these flocks 166,460 chicks were hatched, or 56.7 per cent of the total eggs set. Of these, the mortality for the first three weeks was 10.2 per cent. In 1917, Rettger and Koser-" report on a comparative study of Bacterium pullorum and Bacterium sanguinarium. Their find ings were similar to those reported previously by Smith and TenBroeck. The methyl red test was found to be of value in dis tinguishing the two types in that Bacterium sanguinarium was methyl-red positive. Bacterium sanguinarium is reported as at tacking dextrin and dultite in addition to maltose. Anaerogenic strains of Bacterium pullorum were encountered. The aggluti nation affinities of the tΛvo organisms were found to be similar.
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Limited experiments seemed to show that Bact. pullorum was capable of immunizing rabbits against Baet. sanguinarium, though the latter organism did not reciprocate. Ward and Gallagher, in 1917^^, report on experiments to determine the value of an intradermal test for the detection of carriers of infection. A culture grown for a month and allowed to stand for several weeks, and then carbolized, seemed to give the best results. In their experiments a swelling of the wattle after injection, if observed at the proper time, appeared to be signficant. About 30 hours was stated as being the proper time to make readings. It was found that about 90 per cent of the artificially infected birds reacted to the test and about 6 per cent failed to react although infection was demonstrated on post mortem. In 3 per cent of the cases no reaction occurred and this was in accord with the autopsy. In field experiments the intradermal test failed to agree with the results obtained by agglutination methods, when the reading of the intradermal test was made at 38 hours after the injection. In another field experiment there was agreement in about 70 per cent of the eases. The injection of the material appeared to cause fowls to react to the agglutination test later. In 1917, Goldberg^* reports the fermenting properties of Bact. pullorum and differentiates it from Bact. sanguinarium. His results correspond to those obtained by Rettger and Koser and by Smith and Ten Broeck. The difference in reaction on litmus milk is given as a reliable means of differentiating between the two organisms. Anaerogenic strains of Bacterium pullorum are reported as occurring. In 1917, Krumwiede and Kohn'' point out that both the fowl typhoid and white diarrhea organisms may be differentiated from the human typhoid by their ability to produce acid on rhamnose. Their cross agglutination experiments gave results similar to those obtained by Smith and TenBroeck and by Rettger and Koser. They found, as did Smith and TenBroeck, that the avian strains would not absorb the agglutinins for B. typhosus from a typhoid-immune serum. Hadley, Caldwell, Elkins, and Lambert in 1917,^° report an epidemic in adult fowls due to Bacterium pullorum. Prom the outbreak, which was not attended by high mortality, an organism
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was isolated which resembled Bacterium pullorum, except that it did not produce gas on dextrose and mannite. Eggs laid by some of the affected hens were shown to contain a Bacterium pullorum type which, in some cases, failed to produce gas. Hadley proposes that an Alpha and Beta type of the white diarrhea; organism be recognized, the classification being based on gas production for the Alpha tj'pe. Some strains were found which fluctuated in their ability to produce gas from carbohydrate media. However, the original strain isolated from one of the adult fowls always failed to produce gas. The point is brought out that endogenous infection should not be overlooked. Seherago and Benson, in 1918-19^^, report their results with the intradermal test similar to that used by Ward and Grallagher. From their results, it was concluded that the test was practically worthless as a diagnostic agent in detecting fowls that are carriers of infection. It is also concluded that foreign protein or other substances introduced into the dermis might bring about a reaction similar enough to be mistaken. The application of the intradermal test appeared to cause at least 85 per cent of the birds tested to react positively to the agglutination test later, regardless of their reaction in the original test. Kettger, Kirkpatrick and Card, in 1919^'*, report their results on the infection of adults by introducing cultures into the cloaca and lower part of the oviduct. It was found that such introduction caused the appearance of agglutinins in the blood of some birds. Hens infected in this manner were not shown to transmit the infection to other hens. In one instance, in their first experiment, a generalized infection and death resulted from this method of inoculation. In another instance, an uninoculated hen associating with infected hens appeared to contract the infeetion through the medium of the male as a passive carrier. In a second experiment, infeetion was not spread from inoculated to uninoculated hens, even in a pen in which a male had been placed. It was found that about 25 per cent of the hens which were inoculated into the cloaca became permanent bacillus carriers. In 1919, Mulsow''^, reports on a comparative study of Baet. pullorum and Bacterium sanguinarium. His results were similar to those reported by other investigators. He reports that these
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two oganisms can readily be differentiated from the cholera organism by the action of the latter on litmus milk, its ability to form indol and its fermentation reactions. Mulsow reports that some strains apparently ferment maltose. In 1923, Hitchner'* reported on the influence of the fatty content of the serum on the macroscopic agglutination test. His experiments showed that a high fatty content was usually found in the serum of fowls in heavy production and that a period of starvation preceeding the bleeding removed this interference to a great extent. Starvation for 24 hours gave better results than when the birds were not starved. Starvation for 48 hours caused such a reduction in the fatty content that there was no interference whatever. The starving of birds 36 hours before collecting samples caused sufficient reduction in the fatty content that it was not troublesome. The actual bleeding of fowls without starvation caused no reduction in the egg production, but in birds that had been starved for a period of 36 hours a marked reduction occurred, but after four days there was a slow recovery and normal production Avas reached in the second week. In 1923, Beaudette, Bushnell and Payne^* report on an organism isolated from the unabsorbed yolk of chicks "dead in shell." The strains were almost identical with Bacterium pullorum, except that some of them produced some acid on maltose, a finding previously reported by Mulsow in reference to Bacterium pullorum. The organism was shown to be pathogenic for developing embryos when injected into the egg on the sixth day of incubation. B. typhosus, B. paratyphosus, A and B, also exerted the same action. A. staphylococcus caused the death of the embryo in one of two eggs inoculated. The mortality in inoculated eggs was 100 per cent, as compared with 16.66 per cent in the uninoculated eggs. In the control eggs, one of the eggs used was produced by a hen that had previously produced infected eggs. This egg failed to hatch and on culture yielded a pure strain. I n 1923, Beaudette, Bushnell and Payne^' report on the relation of Bact. pullorum to the hatchability of eggs. It was shown that the average hatch of fertile eggs from infected hens was much lower than the average hatch from uninfected hens. In 1923, Beaudette^" reports the finding of agglutinins for
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SUMMARY
The work that has been done shows, beyond a doubt, that bacillary white diarrhea is a specific infectious disease. The organism which has been named B. puUorum can readily be differentiated from other members of the colon-typhoid group. The hen is the original source of infection and the organism is deposited in the yolk of the eggs from the ovary in which the infection is localized. During the process of incubation, the organism contained in the yolk multiplies rapidly and may cause the death of the embryo in some cases. Should the egg hatch, the chick is infected as soon as it extracts itself from the shell and may show symptoms of the disease very soon. The organism, in chicks thus infected, may be given off in the droppings and infect other chicks not hatched from infected eggs. In this manner, the infection becomes widespread in a flock of baby chicks, even though only a few contracted the infection through the egg. Chicks are most susceptible to infection under forty-eight hours of age. The infection is contracted by eating and drinking food and water contaminated with the droppings of infected chicks. The course of the disease is very short, in most eases, though, some chicks do not die for several weeks. Occasionally, chicks recover the primary infection and mature. However, a large per cent of such recovered chicks will retain the infection in their ovaries and thus constitute permanent carriers. The infection may also be localized elsewhere in the body. Infection of the ovaries in adult fowls can be induced by injecting the organism intravenously. The adult fowl thus having an infected ovary perpetuates the infection. Not all of her eggs will be infected and the per cent of eggs infected varies in different individuals. An adult hen
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Bacterium puUorum in the albumin of eggs laid by infected hens. All but one of the infected birds used in the experiment showed this characteristic. In 1923, Beaudette'^ described a single tube method in making the agglutination test for carriers of infection. The test differed from any previously used in that a drop of serum was used instead of a graduated amount. The drop of serum was placed in 4 cc. of highly diluted antigen which gave a dilution of from 1-80 to 1-100.
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not having infection when j'oung may contract such infection through association with infected hens. Such infection can also be contracted from litter contaminated with the organism. Litter is, in all probability, infected by the droppings of carriers. The mode of infection in adults is probably by way of the digestive tract, though permanent carriers can also be produced by introducing the organism in the cloaca and lower part of the oviduct. Male birds may have infected testes or the infection may be localized elsewhere in the body. The importance of the male as a carrier of infection is probably very small. However, the male might possibly be a passive carrier of infection from one hen to another. It is obvious that in order to eliminate the disease, the hen with infected ovaries must be eliminated from the breeding flock. Such carriers of infection may be determined by egg examinations. The method has not been practical because the organism is eliminated so irregularly that the method requires the examination of a large number of eggs and the cost of such a procedure is therefore prohibitive. An agglutination test has been used and found to give excellent results. An infected bird is one whose blood serum is capable of agglutinating the organism in a dilution of 1-100. A positive agglutination test does not indicate infected ovaries in all cases, as the infection may be localized elsewhere in the body. Furthermore, young fowls niay retain the agglutinins but not the infection, from having had the disease as chicks. An intradermal test has been tried, but the results were so inconsistent that the test was regarded as worthless as a diagnostic means. Agglutinins are also contained in the egg albumin, but because all carriers do not produce such eggs the method cannot be used. The agglutination test has given excellent results, except in some cases where serum samples contain a fatty material that interfers with the test. The proper handling and feeding of fowls prior to the collection of samples has eliminated this difficulty to a great extent. The testing of young birds is not recommended. A single test has resulted in a reduction of the mortality of baby chicks but does not necessarily render the flock free of carriers thereafter, because at the time of testing some fowls may have recently acquired the infection, and time has not elapsed sufiiciently for the
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EEFERENCES
1 RETTGBÏE, L·. F.—Septicaemia Among Young Chickens. New York Medical J o u r n a l , Vol. 71, May 26, 1900, pp. 803-805. 2 RETTGER, L . F.—Septicaemia in Young Chickens. New York Medical J o u r n a l , Vol. 73, F e b r u a r y 16, 1901, pp. 267-268. 3 GRAHAM, C . K.—^Poultry Observations. 1. Causes of Death of Young Chicks. Bulletin No. 44, November, 1906. Storrs A g r i c u l t u r a l Experim e n t Station. * RETTGER, L·. F., AND HARVEY, S . C . — F a t a l Septicaemia in Young Chickens, or "White D i a r r h e a . " J o u r n a l of Medical Research, Vol. 18, N. S. Vol. 13, No. 2, May, 1908, pp. 277-290. 5 M I L K S , H . J.—A P r e l i m i n a r y Report on Some Diseases of Chickens. Bull. No. 108, August, 1908. Louisiana Agricultural E x p e r i m e n t Station. β RETTGER, L . F . — F u r t h e r Studies on F a t a l Septicaemia in Young Chickens, or " W h i t e D i a r r h e a . " J o u r n a l of Medical Research, Vol. 21, No. 1 (N. S. Vol. 16, No. 1 ) , July, 1909, pp. 115-123. 7 RETTGER, L . F . , AND STONEBURN, F . H . — B a c i l l a r y W h i t e D i a r r h e a of Young Chicks. Bulletin No. 60, December, 1909. S t o r r s Agricultural E x p e r i m e n t Station^ s JONES, F . S.—Fatal S e p t i c e m i a or Bacillary W h i t e D i a r r h e a in Young Chickens. Report of t h e New York S t a t e V e t e r i n a r y College for 1910, pp. 111-129. 9 RETTGEE, L . F . , AND STONEBURN, F . H . — B a c i l l a r y W h i t e D i a r r h e a of Young Chicks. (Second R e p o r t ) Bulletin No. 68, April, 1911. S t o r r s Agricultural E x p e r i m e n t Station. 1» GAGE, G . E . — N o t e s on Ovarian Infection w i t h B a c t e r i u m P u l l o r u m ( R e t t g e r ) in t h e Domestic Fowl. J o u r n a l of Medical Research, Vol 24. (N. S. Vol. 19), No. 3, J u n e , 1911.
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appearance of agglutinins in the blood. Furthermore, such freed flocks may acquire infection from contaminated runs and houses after the infected fowls have been eliminated. Annual testing is therefore to be recommended until the flock is shown to be free on two consecutive tests at least. Acute infection with the white diarrhea organism may occur in adult fowls as a septicemia. Such infection may be exogenous or endogenous in origin. The disease may be contracted through the eating of eggs infected with the organism. Some fowls appear to recover from such infection, but they may retain the infection localized in the ovary or other parts of the body and thus be permanent carriers. Endogenous infection may arise from an infection of the ovary. In concluding, it might be said that we have full knowledge regarding the mode of transmission and control of this disease and it remains only a matter of applying this information.
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1* RBTTGEK, L·. F . , KiRKPATRiCK, W M . F . , AND STONEÎBURN, F . H.—Bacil-
lary White D i a r r h e a of Young Chicks. ( T h i r d R e p o r t ) Bulletin No. 74, December, 1912. S t o r r s A g r i c u l t u r a l E x p e r i m e n t Station. 15 BusHNBLi, L. D., AND MATJEIR, 0.—The Use of Milk Cultures of B. Bulgaricus in t h e Prevention and T r e a t m e n t of Bacillary W h i t e Diarrhea. A m e r i c a n V e t e r i n a r y Review, November, 1913, pp. 194-207. 10 GAGE, G . B . , PAIGE, B . H . , AND H Ï L A N D , H . W . — O n t h e Diagnosis of
Infection with B a c t e r i u m P u l l o r u m in t h e Domestic Fowl. Bulletin No. 148, April, 1914. Massachusetts A g r i c u l t u r a l E x p e r i m e n t Station. 17 RETTGER, L . F . — O v a r i a n Infection in t h e Domestic Fowl a n d Direct T r a n s m i s s i o n of Disease to the Offspring. J o u r n a l of E x p e r i m e n t a l Medicine, Vol. 19, No. 6, 1914, pp. 552-561. 18 RETTGER, L·. F., KIRKPATRICK, W M . F . , AND JONES, R . E . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks. A g r i c u l t u r a l E x p e r i m e n t Station.
Bulletin No. 77, J u n e , 1914, S t o r r s
19 RETTGER, L . F . , KIRKPATRICK, W M . P., AND CARD, L . E . — M i l k Feed-
ing a n d I t s Influence on Growth and Mortality. Comparative Study of t h e Value of Sweet and Sour Milk. Bulletin No. 80, April, 1915, S t o r r s Agricultural E x p e r i m e n t Station. 20 S M I T H , T H . , AND T E N BROKCK, CARL.—A Note on t h e R e l a t i o n be-
tween B. P u l l o r u m ( R e t t g e r ) and the Fowl Typhoid Bacillus J o u r n a l of Medical Research, Vol. 31, 1915, pp. 547-555. 21 GAGE, G . E . , AND PAIGE, B . H . — B a c i l l a r y W h i t e D i a r r h e a ium P u l l o r u m Infection) in Young Chicks in Massachusetts. No. 163, August, 1915. Massachusetts A g r i c u l t u r a l E x p e r i m e n t
(Moore), (BacterBulletin Station.
22 RETTGER, L . F . , KIRKPATRICK, W M . F . , AND J O N E S , R . B . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks, (Fifth R é p o r t ) Bulletin No. 85, December, 1915. S t o r r s A g r i c u l t u r a l E x p e r i m e n t Station. 23 GAGE, G . B . , AND MARTIN,-JAMES.^-Notes on t h e Histo-Pathology of t h e I n t e s t i n e s in Young Chicks Infected w i t h B a c t e r i u m Pullorum. J o u r n a l of Medical Research, Vol. 34, 1916, pp. 149-156. 2* RETTGER, L . F . , H i n x , T. G., AND STTJRGES, W M . S.—Feeding Experim e n t s with B a c t e r i u m P u l l o r u m . T h e Toxicity of Infected Eggs. Journal of E x p e r i m e n t a l Medicine, Vol. 23, No. 4, April, 1916, pp. 475-489. 25 RETTGER, L . F . , KIRKPATRICK, W M . F . , AND J O N E S , R . E . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks. ( S i x t h R e p o r t ) Bulletin No. 88, Oc-
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11 JONES, F . S . — F u r t h e r Studies on Bacillary W h i t e D i a r r h e a in Young Chickens. Report of t h e New York S t a t e V e t e r i n a r y College for 1911, pp. 69-88. 12 JONES, F . S.—An Outbreak of an Acute Disease in Adult Fowls Due t o B a c t e r i u m P u l l o r u m . Report of t h e New York S t a t e V e t e r i n a r y College for 1911-12, p p . 140-148. J o u r n a l of Medical Research, Vol. 27, 1913, pp. 471-479. 13 JONES, F . S · — T h e Value of t h e Macroscopic Agglutination Test in Detecting Fowls t h a t a r e Hairboring B a c t e r i u m P u l l o r u m . Report of the New York S t a t e V e t e r i n a r y College for 1911-12, pp. 149-158. J o u r n a l of Medical Research, Vol. 27, No. 4, 1913, pp. 481-495.
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-8 KRUMWIEDB, C , J R . , AND K O K N , L·. A.—Studies on t h e P a r a t y p h o i d -
E n t e r i t i d i s Group. IV. T h e pifferentation of t h e Members of t h e Paratyphoid-Enteritidis Group from B. Typhosus w i t h Special Reference to the Anserogenic S t r a i n s a n d Observations on t h e F e r m e n t a t i v e Charac teristics of • t h e Avian Types. J o u r n a l of Medical Research, Vol. 36, 1917, p. 509. 30 H A D L E Y , P . B . , C A L D W E L L , D . W . , E L K I N S , M . W . , A N D L A M B E R T , D . J .
—Infections Caused by B a c t e r i u m P u l l o r u m in Adult Fowls. Bulletin 172, November, 1917. A g r i c u l t u r a l E x p e r i m e n t Station of t h e Rhode Island State College. 31 SCHEBAGO, M . , AND BENSON, J. P . — E x p e r i m e n t s on t h e I n t r a d e r m a l
Test for B a c t e r i u m P u l l o r u m . Report of t h e New York State Veteri n a r y College for 1918-1919, p p . 183-191. Cornell Veterinarian, Vol. 9, 1919, p p . 111-119. 3-' RETTGEB, L . F . , KntKPATEicK, W M . F . , AND CARD, L . E . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks. (Seventh R e p o r t ) Bulletin No. 101, J a n u a r y , 1919. Storrs A g r i c u l t u r a l E x p e r i m e n t Station. 33 MuLsow, F . W.—A Comparative S t u d y of B . P u l l o r u m and B. San g u i n a r i u m . J o u r n a l of Infectious Diseases, Vol. 25, No. 2, August, 1919. 3* BEAUDETTB, F . R . , B U S H N E X L , L . D . , AND P A Y N E , L . F . — S t u d y of a n
Organism Resembling Bact. P u l l o r u m from Unabsorbed Yolk of Chicks "Dead in Shell." J o u r n a l of Infecious Diseases, Vol. 32, No. 2, Feb r u a r y , 1923, pp. 124-132.* 35 BEAUDETTE, F . R . , B U S H N E L L , L . D . , AND P A Y N E , L . F . — R e l a t i o n of
B a c t e r i u m P u l l o r u m to H a t c h a b i l i t y of Eggs. J o u r n a l of Infectious Diseases, Vol. 33, No. 4, October, 1923, pp. 331-337. 36 BEAUDETTE, F . R . — A g g l u t i n i n s for Bacterium P u l l o r u m in H e n s ' Eggs. J o u r n a l of Immunology, Vol. 8, No. 6, November, 1923, pp. 493497. 37 BEALTDETTE, F . R.—Single Tube Method for D e t e r m i n i n g C a r r i e r s of B a c t e r i u m P u l l o r u m . J o u r n a l of t h e A m e r i c a n Veterinary Medimal As sociation, Vol. 64, (N. S. 17) No. 2, November 1923, pp. 225-227. 38 HrrcHNiai, E . R.—The Macroscopic Agglutination Test as Influ enced by t h e F a t t y Content of t h e Blood Serum. J o u r . Amer. Vet. Med. Association, Vol. 63, No. 6 ( N . S. Vol. 1 6 ) , September, 1923, p p . 759-763.
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tober, 1916. S t o r r s Agricultural E x p e r i m e n t Station. 2β RETTGEK, L . P . , AND KOSEK, S . A.—A Comparative Study of Bac t e r i u m P u l l o r u m ( R e t t g e r ) a n d Bacterium S a n g u i n a r i u m (Moore). J o u r n a l of Medical Research, Vol. 35 (N. S. Vol. 3 0 ) , No. 3, J a n u a r y , 1917, pp. 443-458. -1 WABD, A. R., AND GAii.AGHEB, B. A.—An I n t r a d e r m a l Test for Bac t e r i u m P u l l o r u m Infection in Fowls. Bulletin No. 517, F e b r u a r y , 1917. U. S. D. Α., Β. Α. I. 28 GoLDBEBG, S. Α.—A Study of t h e F e r m e n t i n g P r o p e r t i e s of Bac t e r i u m P u l l o r u m ( R e t t g e r ) a n d B a c t e r i u m S a n g u i n a r i u m (Moore). J o u r n a l A m e r i c a n Veterinary Medical Association, Vol 51 (N. S. Vol. 4), 1917, pp. 203-210.
AUGUST-SEPTEMBER, 1925
No. 6
BACILLARY WHITE DIARRHEA* F. R. BEAtTDETTB New Jersey Agricultural Experiment Station
* Paper No. 198 of the Journal Series, New Jersey Agriculturay Experiment Station, Department of Poultry Husbandry.
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Since bacillary white diarrhea Avas described by Rettger in 1900, its presence has been determined in every part of the United States. The disease has been found in Europe and future investigation will no doubt show that it is present in every locality where poultry raising thrives. The ease with which this disease can be introduced into a new locality makes its spread all the more possible. The economic importance of bacillary white diarrhea has attracted many to its investigation and the information thus derived has been plentitful. In fact, it can be said that more is known about this disease than any other malady that affects the common fowl. In order to appreciate this fact, it might be well to scan the various contributions that have elucidated the nature of this disease. Rettger,^ in 1900, was the first to describe the organism that is responsible for white diarrhea, though at that time the disease was described as a septicaemia and its relation to what is now known as bacillary white diarrhea was probably not suspected. On this occasion his observations were confined to a brood of seventeen chicks, all but three of which became sick. Of the fourteen sick chicks eleven died and three finally recovered, though they remained stunted in growth for two months or more, after Avhich they grew rapidly again. The symptoms and post mortem findings of the disease were described quite fully. From the livers of the chicks, and in one case from the spleen, a small bacillus was isolated and described. The organism was reported as being motile, an observation which was later found to be incorrect by the Same worker.
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In order to determine the pathogenicity of this organism, four chicks ranging in age from two to four weeks were inoculated subeutaneously with 0.5 cc. of a twenty-four hour bouillon cul ture. All of the chicks became sick and two died. A third was killed after it had showed signs of recovery. From the two chicks that had died and from the third that was killed the original or ganism was isolated from the livers and in one instance from the lungs and kidneys. Mention was made of the fact that the mother hen as well as other grown chicks were apparently im mune, and it is to be noted also that in the inoculation experi ment the younger chicks died, whereas the older ones recovered. In 1901, Rettger- again reports his findings in regard to a similar disease of larger proportions occurring at Winona Lake. In a lot of forty chicks (four broods), thirty-five died while between one and four weeks old. In another brood of fifteen the disease appeared before the chicks were a week old. Later, two other broods of thirty-three chicks became affected and in less than three weeks all but ten had died. The disease next ap peared on two adjoining farms where 80 per cent of the chicks died that contracted the disease. The organisms isolated from the livers of the chicks at this time corresponded with that described previously by the same author. By means of this organism he Avas again able to cause the death of two chicks (two weeks old) by subcutaneous injection. The economic importance of the disease as manifested by its epi demic nature was noted at this time. In 1906, Graham^ reports a disease of young chicks at the Storrs Station. The disease appeared in a February hatch of 400 chicks and it is stated that nearly all of them died. A January hatch of 150 chicks and a March hatch of 500 chicks were apparently not affected. The disease was undoubtedly white diarrhea and this outbreak probably marks its introduc tion into this Station, where it was thoroughly investigated in later years. In 1908, Rettger and Harvey* report a third epidemic that occurred in Connecticut in 1907. In this outbreak one hundred and forty-six chicks were exposed to the disease and eighty-four or about sixty per cent died. The bacillus isolated resembled the organisms obtained from the tAvo previous epidemics. In
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describing the organism it is stated to be non-motile, whereas it was previously said to be motile. The cultural features of the organism are again reported and its ability slightly to acidify litmus milk is noted for the first time. The fermentation reactions of the organism are also reported, for the first time. Two guinea pigs inoculated with 1.5 cc. subcutaneously and 5.5 cc. intraperitoneally with broth cultures respectively proved refractory. It was found that a rabbit was not killed by three injections of living culture after it had been immunized with several injections of dead culture. The disease was produced for the first time by feeding the organism to two chicks ten and eight days old. Death occurred on the fourth and nineteenth day respectively. It was noted on this occasion that the serum of a chick that had been injected with killed cultures would agglutinate the homologous organism. The same reaction was also noted in connection with the serum of an immunized rabbit. The identity of this disease with what is commonly known as white diarrhea is brought out. The causative organism is placed for the first time in the colon-typhoid group. The source of infection is still unsolved, though it was brought out that transmission of the disease from chick to chick might take place through the medium of contaminated food and drinking water. Whether the egg is infected before it is laid, during incubation, or not until the chick is hatched, are yet unsolved problems. In 1908, Milks'^ describes an outbreak that came to his attention in 1907. The disease was confined to chicks under five or six weeks old. The symptoms and post mortem lesions described were undoubtedly those of white diarrhea. An organism was isolated whose cultural features and fermentive powers are identical with the organism described by Rettger. The occurrence of grayishwhite areas on the heart is noted for the first time. In 1909, Rettger* again describes the disease and calls attention to the almost constant presence of an unabsorbed yolk in chicks dead of this disease. Mention is also made of a semi-solid or cheesy plug found in the ceca. This lesion was noted by Milks in 1908. The organism is named Bacterium pullorum and methods are given for its isolation and identification. The oc-
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currence of anaerogenic strains of the organism is noted for the first time. The organism is reported as being isolated from the crop, stomach and intestines, as well as from the unabsorbed yolk. Experiments showed the organism to be particularly suscepti ble to the action of lactic acid, and it was suggested that the feeding of sour milk might have beneficial results. That the egg is suspected as a source of infection, is evidenced by the fact that over five dozen eggs were examined. However, these examina tions were negative in every ease. In 1909, Rettger and Stoneburn' record extensive observa tions made at the Storrs Station. White diarrhea had ap parently been prevalent on this plant for several seasons and it was deemed advisable to dispose of a greater part of the breed ing stock. On dressing the birds for market they were noted to be in poor condition and several showed pathological ovaries. The first hatch of the season came off in February 1909. In a lot of 250 chicks a few were reported as dying from "White diarrhea." The brooders were again filled in March, and this time serious losses were sustained. Specimens were examined, and the specific organism was isolated from the internal organs. Because a number of Rhode Island Red hens seemed to be par ticularly affected, it was decided to make a study of their eggs. The shells of twelve eggs were examined with negative results. The yolks of eighty-six incubated eggs yielded a pure culture in eight instances, five were questionable. The eggs were stated to have come from a number of Λ'arieties, but the organism was ob tained from eggs laid by Rhode Island Red hens in seven in stances and only in one egg laid by a Buff Orpington. The yolks of twelve fresh Rhode Island Red eggs were examined and the organism was recovered from one. The ovaries of twenty-three hens were next examined and twenty-one showed pathological changes. Cultures were taken from twenty of the ovaries and the organism was obtained in nine instances. Six other Rhode Island Red hens were examined and in every instance where the ova were pathological the organism was recovered. Prom these findings it was concluded that the hen is the original source of infection. The normal mortality in 280 chicks from the Station stock
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was found to be about 13 per cent. A few drops of water suspen sion of B. puUorum given to each of fif tyincubator hatched chicks caused the disease to develope in a large number and thirty-four or 68 per cent died. In another infection experiment where bouillon cultures were used, the mortality at the end of eight weeks was 47 per cent for the infected chicks and 17 per cent for the controls. The organism was recovered from about 75 per cent of the chicks that were infected. In a third experiment where the culture was supplied in the drinking water, a big dif ference Avas noted in the mortality of the infected and control lots. In 1910, Jones'* states that his attention was first called to this disease in 1910. On this occasion six one week old chicks were examined and the organism recovered. The mortality in this out break was over 90 per cent and the year previous about 1800 chicks had died apparently from the same disease. The disease appeared to have been present in this flock to some extent in 1908. Of 104 chicks produced on this plant and shipped twentyfive miles, thirty-eight remained well at the end of t ^ o weeks. In another lot of 100 chicks that were shipped, fifty-five had died. A lot of twenty eggs injected with 0.1 cc. of Bacterium pullorum failed.to hatch, although the embryo developed for about fifteen days. Another lot similarly injected with sterile bouillon produced fifteen chicks. Eggs smeared with cultures of Bacterium pullorum and then incubated produced chicks, none of which died of the disease. In 1911, Rettger and Stoneburn" published their second report of work done at the Storrs Station. Cultures of Bacterium pul lorum fed to chicks 12 hours old caused a mortality of 71 per cent, as compared with 4 per cent in the control lots. The in fected chicks Λvere stated to be at least 25 per cent lighter in weight than the uninfected chicks. The teehnic of examining fresh eggs is described wherein the whole yolk is added to a quantity of broth and incubated 72 hours. From this, tubes are streaked to determine the presence of Bacterium pullorum. It was noted that infected hens seem to be poor layers, especially during their second year. Also, that chicks hatched in winter and in early spring were comparatively free of the disease. The question of hens contracting infection as adults is still un-
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solved. The male bird as a factor in transmission is also unsolved. Testis have been examined, but with negative results. A cycle of infection is represented for the first time. It is to be recalled, however, that the way in which hens become infected is not as yet definitely established. In 1911, Gage^" reports the isolation of Bacterium pullorum from the ovaries of hens. The organism was isolated from twelve different ova of the same hen and the pathogenicity of these strains tested on baby chicks. Eighty-two chicks were used. Sixty-two were inoculated, thirty by subcutaneous injection and thirty-two were fed two drops of bouillon culture. The remaining twenty chicks were held as controls, and at the end of six weeks, all of the controls were alive and in good condition. Of the sixty-two inoculated and fed chicks, sixty died and the organism was recovered in every case. In 1911, Jones^^ showed that young chicks which recovered from the primary infection might retain the infection in their ovaries, as evidenced by examinations of their eggs and ovaries. It was also shown that ovarian infection could be induced by intravenous injection of the organism. For detecting carriers of infection, Jones suggested the use of the agglutination test. In 1911-12, Jones" reports an epidemic in adult fowls due to Bacterium pullorum. In this instance a poultryman who had no trouble from this disease incubated eggs for a neighbor who had very poor success. The eggs produced a poor hatch and some of the incubated eggs were fed to adult hens. In sixteen days after the eggs were fed losses occurred, and in all, fifty fowls died. The organism was recovered from the pericardium, liver, spleen and ovaries of the dead hens. This farm had never had white diarrhea in baby chicks, but after the feeding of the incubated eggs, the chicks produced from this stock showed the disease, indicating that ovarian infection was undoubtedly produced by feeding of incubated infected eggs. "The infection in adults is characterized by necrotic foci in the liver, spleen and pancreas and large necrotic nodules in the heart muscle. The presence of a fibrinous exudate on the capsule of the liver and spleen and on the pericardium and heart was a somewhat constant lesion." The occurrence of nodules on the heart was first reported by Milks in 1908.
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In 1911-12, Jones^^ reports on the value of the macroscopic agglutination test in detecting fowls that are harboring ovarian infection. Of the twenty-one fowls found to give: a positive reaction by this test, twenty were shown to be carriers by bacteriological examination of the ovaries. I t was stated that probably the remaining one Avould have been found to be infected had more extensive searches been made. Unheated test fluid was found to give a little better result than the heated antigen. The methods of collecting blood samples, the making of test fluid and making the test are also described. In 1912, Rettger, Kirkpatrick and Stoneburn^* report further experiments. To determine the age of susceptibility, five lots were used containing twenty-six chicks each. One lot was not infected and remained as a control and the mortality was 11.5 per cent in this lot. Of the chicks infected at 36 hours, 33.5 per cent died ; 23.33 per cent of the chicks infected at 60 hours died ; 19.25 per cent loss was sustained in those infected at 84 hours and only 7.7 per cent of the chicks died that were infected at the age of 108 hours. In a second experiment 300 chicks were used and divided equally into six lots of 50 chicks each. 16.66 per cent of the controls died ; 72.11 per cent of those infected at 24 hours died ; 88 per cent infected at 48 hours died ; 30 pei· cent of those infected at 72 hours succumbed ; 20 per cent of the lot infected at 96 hours died and the mortality in the lot infected at 120 hours was 16.0 per cent. It was concluded from these experiments that chicks are most susceptible up to forty-eight hours after hatching. Fourteen hens were placed together, seven of which were known to be carriers and seven were known not to be infected by egg examinations. The birds were allowed to associate together for about two years and at the end only four of the originally uninfected hens remained alive. About eight months after the beginning of the experiment. Bacterium pullorum was found in one of the eggs of the uninfected birds, a little later two more infected eggs were found and it was determined that all three eggs came from the same hen. Sixteen months later, an infected egg was found from one of the other hens and a week later a second egg from thé same hen was found to contain the organism. Still later, an infected egg was laid by the third hen and on post mortem examination all four hens were
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found to have pathological ovaries. The experiment showed that infection might be contracted by association with infected hens. In order to determine whether hens could contract infection from infected litter, six pens were maintained containing 8 hens each, three of the pens served as controls. Bacterium puUorum was sprinkled on the litter of three of the pens two or three times a week. Eight White Leghorn and forty Rhode Island Red pullets were used and the experiment was carried on for one year. At the end of the year, the 24 pullets serving as controls had not laid an infected egg and none of their ovaries were found to be pathological on post mortem examination. Of the twentyfour birds in infected pens, eight had produced infected eggs and a ninth bird showed pathological ovaries on post mortem. The experiment proved beyond a doubt that infection of the ovary could be contracted from infected litter. Experiments were carried on to determine the value of sour milk as a preventive. A large number of chicks were used in several trials. In every ease, the chicks which had received sour milk showed a much lower mortality than did the infected chicks which had not received sour milk. Chicks fed sour milk were found to make more rapid gains. In 1913, Bushnell and Maurer^" report on the value of milk cultures of B. bulgaricus as a preventive for this disease. In their experiments it was shown that such milk exerted a beneficial action in that when administered 40 hours before infection, the mortality was much lower than in check pens which did not receive the milk. The feeding of B. bulgaricus milk 40 hours after infection with Bacterium pullorum prevented as high a mortality as was experienced in the check pens. In 1914, Gage, Paige and Hyland" confirmed previous observations made by Rettger, that the organism was more readily isolated from incubated eggs than from fresh eggs. The technie used was essentially the same as that described by Rettger. Their examination showed that from 4 to 21 eggs must be examined, before the organism was recovered in the experimental birds used in their work. The laying period covered fom 6 to 58 days. It was shown that hens inoculated some two years previously with a culture of Bacterium pullorum intravenously resulted in an ovarian infection.
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In 1915, Rettger, Kirkpatrick and Card^" reported results that indicated no difference in the results of feeding sweet or sour milk. Milk soured with B. bulgarieus gave no advantage over that soured naturally. The beneficial results are explained on the basis that milk feeding increased growth and vitality to such an extent that the resistance of the chick was increased and hence made it more immune to infection. Milk feeding seemed to prevent mortality from all causes. It was concluded from their
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The eggs from a number of hens were examined and it was demonstrated that a negative egg record also gave negative results on autopsy, whereas all but two hens yielded cultures on autopsy that had given a positive agglutination reaction. It is explained that these two probably retained only the agglutinin from the primary infection. Polyvalent test fluid was found,to give better results than a monovalent antigen. In 1914, Rettger"^' reviews direct transmission of the disease to the offspring. Again in 1914, Rettger, Kirkpatrick and Jones^' positively proved that chicks having the infection when young and recovering from it, retained in the ovaries a localized infection in some instances. Chicks w^ere infected when young and some recovered, when tested at the age of one year a number reacted positively to the agglutination test and on autopsy these showed pathological ovaries. Of 138 chicks growing to maturity, 88 were infected when quite young and 57 were held as controls. Of the 88 infected when young, a positive agglutination test was obtained on 21, showing that over 25 per cent retained the infection localized in the ovary. One of the 57 controls gave a positive test and also showed pathological ovaries. This was probably a contact case, as it was in close proximity to some of the infected birds. The macroscopic agglutination test was found to agree with post mortem examination. Evidence was presented which seemed to show that agglutinins would appear in the serum of fowls fed a ration infected with the organism. In one instance, a iowl gave a positive serum test, but on post mortem examination the ovaries were found to be normal. However, a thoracic tumor was found and from this a pure culture of Bacterium puUorum was isolated.
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study that milk as a food was not of value because of the acids which it might contain, but upon the natural constituents of the milk itself. In 1915, Smith and TenBroeck^" present data on a comparative study of Bact. pullorum and Bact. sanguinarium. They conclude that the two organisms are different. It was shown that Bact. pullorum could be distinguished from Bact. sanguinarium because the former was able to produce gas on dextrose and mannite, whereas the latter failed in this respect. The ability of Baet. sanguinarium to produce acid on maltose also distinguished it from the white diarrhea organism. In their «studies of the agglutination affinities it was pointed out that the white diarrhea organism was closely related to the human typhoid bacillus and the fowl typhoid organism in its serum reactions.. Rabbits were killed with bacterial free filtrates of 5 and 15 day-old cultures of the white diarrhea organism. The animals died from what appeared to be a toxic action of the organism. In 1915, Gage and Paige-^ show, by chick examination, that bacillary white diarrhea is very prevalent and widespread in Massachusetts. Various breeds were tested by the agglutination method and the range of infection varied from 0 per cent to over 50 per cent in the flocks tested. It was stated that reacting fowls were found which originated in Europe. Fowls whose serum caused agglutination in a 1-100 dilution were considered as infected. Rettger, Kirkpatrick and Jones in 1915^^^ give the results of testing 14,617 fowls contained in 107 different flocks. Of this number, 13,831 were hens and 1417 or 10.24 per cent were found to be infected while only 23 or 2.9 per cent of the 786 males tested reacted. The per cent of infection in both males and females was 9.85 per cent. Only 4.3 per cent of the Leghorns were found to be infected; 12.2 per cent of the Rhode Island Reds gave a positive test ; 13.7 per cent of the Plymouth Rocks were found to be infected and the Wyandottes reacted to the extent of 23.9 per cent, while the miscellaneous breeds showed a 17.1 per cent infection. The per cent of infection was found to be greatest in large flocks. Eleven of the 23 reacting males were killed and examined for the presence of Bacterium pullorum. In the testes of two of the
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males the organism was recovered in pure culture. In three of the males the organism was isolated from the pericardial sac. In a fourth male both the testes and pericardial sac were found to be infected. The presence of agglutinins in the six males from which the organism was not recovered is explained on the basis that they merely retained the agglutinins from the infection while young. Thirteen flocks were retested and in four cases no reactors were found on second test. The per cent of infection varied in the other flocks from 0.6 per cent to 25.7 per cent, though in every case the per cent of infected birds on second test was less than was found on the first test. In 1916, Gage and Martin'^' report studies on the histopathology on the intestines of chicks dead of white diarrhea. In a lot of 63 chicks, 53 were injected with a mixture of three strains of Baet. puUorum. Of the inoculated chicks, 26 died in from 4 to 11 days and 27 recovered. The control chicks did not die and grew normally. The occurrence of death at different ages afforded a variety of material with which to study the changes in the intestine. Sections of acute cases showed a hyperemia and catarrhal exudation with desquamation of the epithelium. In some cases hemorrhage was in evidence. Mononuclear lymphocytes were found to be numerous. In more chronic cases regeneration of the epithelium had started and proliferating fibroblasts were seen. Mucus and white cells were abundant. In chronic eases a thickening of the intestine was noted. In 1916, Rettger, Hull and Sturges^* report results with feeding experiments with Bact. puUorum to rabbits, kittens, guinea pigs and white rats. Young rabbits were found to be most susceptible and death usually took place. In kittens symptoms of severe food-poisoning were noted. The possibility of infection to man from eating infected eggs improperly cooked is pointed out. The resistance of the organism in eggs subjected to frying, coddling, boiling, poaching and scrambling was also studied. It was found that boiling for four minutes did not kill the organism in all cases. Frying of eggs hard on one side also failed to render them sterile in all cases. Prying on both sides was found to be effective as was scrambling and poaching. Rettger, Kirkpatrick and Jones in 1916^^ give a progress re-
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port on the testing of fowls for carriers of white diarrhea. In all, 7799 birds were tested, 6358 of which were retested from the year preceeding but coming from infected flocks at that time. One hundred and fiftj'^-seven males M^ere tested, 96 of which were tested for the first time. All males reacted negatively. The distribution of infection among the various breeds was found to be as follows:—Rhode Island Reds, 11.9 per cent; White Leg horns, 3.4 per cent ; Wyandottes, 10.3 per cent ; Plymouth Rocks, 11.7 per cent; Orpingtons, 31.7 per cent; FaveroUes, 22.2 per cent and miscellaneous breeds 23.2 per cent. Of all breeds, 8.2 per cent were found to be infected. Of the 78 flocks tested, 25 were found to be free of infection. Four of the Leghorn flocks were found to be free on retest. Of the seven Rhode Islajid Red flocks, five showed infection on second test. In all cases, the second test showed a lower per cent infection than the first test. All three Barred Rock flocks showed reactors on second test and in 5 Wyandotte flocks the number of carriers found on second test was much lower than found on first test. The infection was reduced from 3.5 per cent to 0.0 per cent in Leghorn flocks; in Rhode Island Red flocks from 11.25 per cent to 1.8 per cent; in Barred Rocks from 22.8 per cent to 7.6 per cent ; in Wyandottes from 20.8 to 14.5 per cent ; the reduction in all flocks was from 13.16 per cent on first test to 6.37 per cent on second test. The total number of fowls tested in the two years was stated to be 21,317 with a 9.3 per cent average infection. Of the 1037 males tested, 2.1 per cent reacted. The per cent of infection in various flocks ranged from 0.0 per cent to 56.3 per cent. It was stated also that from these flocks 166,460 chicks were hatched, or 56.7 per cent of the total eggs set. Of these, the mortality for the first three weeks was 10.2 per cent. In 1917, Rettger and Koser-" report on a comparative study of Bacterium pullorum and Bacterium sanguinarium. Their find ings were similar to those reported previously by Smith and TenBroeck. The methyl red test was found to be of value in dis tinguishing the two types in that Bacterium sanguinarium was methyl-red positive. Bacterium sanguinarium is reported as at tacking dextrin and dultite in addition to maltose. Anaerogenic strains of Bacterium pullorum were encountered. The aggluti nation affinities of the tΛvo organisms were found to be similar.
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Limited experiments seemed to show that Bact. pullorum was capable of immunizing rabbits against Baet. sanguinarium, though the latter organism did not reciprocate. Ward and Gallagher, in 1917^^, report on experiments to determine the value of an intradermal test for the detection of carriers of infection. A culture grown for a month and allowed to stand for several weeks, and then carbolized, seemed to give the best results. In their experiments a swelling of the wattle after injection, if observed at the proper time, appeared to be signficant. About 30 hours was stated as being the proper time to make readings. It was found that about 90 per cent of the artificially infected birds reacted to the test and about 6 per cent failed to react although infection was demonstrated on post mortem. In 3 per cent of the cases no reaction occurred and this was in accord with the autopsy. In field experiments the intradermal test failed to agree with the results obtained by agglutination methods, when the reading of the intradermal test was made at 38 hours after the injection. In another field experiment there was agreement in about 70 per cent of the eases. The injection of the material appeared to cause fowls to react to the agglutination test later. In 1917, Goldberg^* reports the fermenting properties of Bact. pullorum and differentiates it from Bact. sanguinarium. His results correspond to those obtained by Rettger and Koser and by Smith and Ten Broeck. The difference in reaction on litmus milk is given as a reliable means of differentiating between the two organisms. Anaerogenic strains of Bacterium pullorum are reported as occurring. In 1917, Krumwiede and Kohn'' point out that both the fowl typhoid and white diarrhea organisms may be differentiated from the human typhoid by their ability to produce acid on rhamnose. Their cross agglutination experiments gave results similar to those obtained by Smith and TenBroeck and by Rettger and Koser. They found, as did Smith and TenBroeck, that the avian strains would not absorb the agglutinins for B. typhosus from a typhoid-immune serum. Hadley, Caldwell, Elkins, and Lambert in 1917,^° report an epidemic in adult fowls due to Bacterium pullorum. Prom the outbreak, which was not attended by high mortality, an organism
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was isolated which resembled Bacterium pullorum, except that it did not produce gas on dextrose and mannite. Eggs laid by some of the affected hens were shown to contain a Bacterium pullorum type which, in some cases, failed to produce gas. Hadley proposes that an Alpha and Beta type of the white diarrhea; organism be recognized, the classification being based on gas production for the Alpha tj'pe. Some strains were found which fluctuated in their ability to produce gas from carbohydrate media. However, the original strain isolated from one of the adult fowls always failed to produce gas. The point is brought out that endogenous infection should not be overlooked. Seherago and Benson, in 1918-19^^, report their results with the intradermal test similar to that used by Ward and Grallagher. From their results, it was concluded that the test was practically worthless as a diagnostic agent in detecting fowls that are carriers of infection. It is also concluded that foreign protein or other substances introduced into the dermis might bring about a reaction similar enough to be mistaken. The application of the intradermal test appeared to cause at least 85 per cent of the birds tested to react positively to the agglutination test later, regardless of their reaction in the original test. Kettger, Kirkpatrick and Card, in 1919^'*, report their results on the infection of adults by introducing cultures into the cloaca and lower part of the oviduct. It was found that such introduction caused the appearance of agglutinins in the blood of some birds. Hens infected in this manner were not shown to transmit the infection to other hens. In one instance, in their first experiment, a generalized infection and death resulted from this method of inoculation. In another instance, an uninoculated hen associating with infected hens appeared to contract the infeetion through the medium of the male as a passive carrier. In a second experiment, infeetion was not spread from inoculated to uninoculated hens, even in a pen in which a male had been placed. It was found that about 25 per cent of the hens which were inoculated into the cloaca became permanent bacillus carriers. In 1919, Mulsow''^, reports on a comparative study of Baet. pullorum and Bacterium sanguinarium. His results were similar to those reported by other investigators. He reports that these
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two oganisms can readily be differentiated from the cholera organism by the action of the latter on litmus milk, its ability to form indol and its fermentation reactions. Mulsow reports that some strains apparently ferment maltose. In 1923, Hitchner'* reported on the influence of the fatty content of the serum on the macroscopic agglutination test. His experiments showed that a high fatty content was usually found in the serum of fowls in heavy production and that a period of starvation preceeding the bleeding removed this interference to a great extent. Starvation for 24 hours gave better results than when the birds were not starved. Starvation for 48 hours caused such a reduction in the fatty content that there was no interference whatever. The starving of birds 36 hours before collecting samples caused sufficient reduction in the fatty content that it was not troublesome. The actual bleeding of fowls without starvation caused no reduction in the egg production, but in birds that had been starved for a period of 36 hours a marked reduction occurred, but after four days there was a slow recovery and normal production Avas reached in the second week. In 1923, Beaudette, Bushnell and Payne^* report on an organism isolated from the unabsorbed yolk of chicks "dead in shell." The strains were almost identical with Bacterium pullorum, except that some of them produced some acid on maltose, a finding previously reported by Mulsow in reference to Bacterium pullorum. The organism was shown to be pathogenic for developing embryos when injected into the egg on the sixth day of incubation. B. typhosus, B. paratyphosus, A and B, also exerted the same action. A. staphylococcus caused the death of the embryo in one of two eggs inoculated. The mortality in inoculated eggs was 100 per cent, as compared with 16.66 per cent in the uninoculated eggs. In the control eggs, one of the eggs used was produced by a hen that had previously produced infected eggs. This egg failed to hatch and on culture yielded a pure strain. I n 1923, Beaudette, Bushnell and Payne^' report on the relation of Bact. pullorum to the hatchability of eggs. It was shown that the average hatch of fertile eggs from infected hens was much lower than the average hatch from uninfected hens. In 1923, Beaudette^" reports the finding of agglutinins for
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SUMMARY
The work that has been done shows, beyond a doubt, that bacillary white diarrhea is a specific infectious disease. The organism which has been named B. puUorum can readily be differentiated from other members of the colon-typhoid group. The hen is the original source of infection and the organism is deposited in the yolk of the eggs from the ovary in which the infection is localized. During the process of incubation, the organism contained in the yolk multiplies rapidly and may cause the death of the embryo in some cases. Should the egg hatch, the chick is infected as soon as it extracts itself from the shell and may show symptoms of the disease very soon. The organism, in chicks thus infected, may be given off in the droppings and infect other chicks not hatched from infected eggs. In this manner, the infection becomes widespread in a flock of baby chicks, even though only a few contracted the infection through the egg. Chicks are most susceptible to infection under forty-eight hours of age. The infection is contracted by eating and drinking food and water contaminated with the droppings of infected chicks. The course of the disease is very short, in most eases, though, some chicks do not die for several weeks. Occasionally, chicks recover the primary infection and mature. However, a large per cent of such recovered chicks will retain the infection in their ovaries and thus constitute permanent carriers. The infection may also be localized elsewhere in the body. Infection of the ovaries in adult fowls can be induced by injecting the organism intravenously. The adult fowl thus having an infected ovary perpetuates the infection. Not all of her eggs will be infected and the per cent of eggs infected varies in different individuals. An adult hen
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Bacterium puUorum in the albumin of eggs laid by infected hens. All but one of the infected birds used in the experiment showed this characteristic. In 1923, Beaudette'^ described a single tube method in making the agglutination test for carriers of infection. The test differed from any previously used in that a drop of serum was used instead of a graduated amount. The drop of serum was placed in 4 cc. of highly diluted antigen which gave a dilution of from 1-80 to 1-100.
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not having infection when j'oung may contract such infection through association with infected hens. Such infection can also be contracted from litter contaminated with the organism. Litter is, in all probability, infected by the droppings of carriers. The mode of infection in adults is probably by way of the digestive tract, though permanent carriers can also be produced by introducing the organism in the cloaca and lower part of the oviduct. Male birds may have infected testes or the infection may be localized elsewhere in the body. The importance of the male as a carrier of infection is probably very small. However, the male might possibly be a passive carrier of infection from one hen to another. It is obvious that in order to eliminate the disease, the hen with infected ovaries must be eliminated from the breeding flock. Such carriers of infection may be determined by egg examinations. The method has not been practical because the organism is eliminated so irregularly that the method requires the examination of a large number of eggs and the cost of such a procedure is therefore prohibitive. An agglutination test has been used and found to give excellent results. An infected bird is one whose blood serum is capable of agglutinating the organism in a dilution of 1-100. A positive agglutination test does not indicate infected ovaries in all cases, as the infection may be localized elsewhere in the body. Furthermore, young fowls niay retain the agglutinins but not the infection, from having had the disease as chicks. An intradermal test has been tried, but the results were so inconsistent that the test was regarded as worthless as a diagnostic means. Agglutinins are also contained in the egg albumin, but because all carriers do not produce such eggs the method cannot be used. The agglutination test has given excellent results, except in some cases where serum samples contain a fatty material that interfers with the test. The proper handling and feeding of fowls prior to the collection of samples has eliminated this difficulty to a great extent. The testing of young birds is not recommended. A single test has resulted in a reduction of the mortality of baby chicks but does not necessarily render the flock free of carriers thereafter, because at the time of testing some fowls may have recently acquired the infection, and time has not elapsed sufiiciently for the
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EEFERENCES
1 RETTGBÏE, L·. F.—Septicaemia Among Young Chickens. New York Medical J o u r n a l , Vol. 71, May 26, 1900, pp. 803-805. 2 RETTGER, L . F.—Septicaemia in Young Chickens. New York Medical J o u r n a l , Vol. 73, F e b r u a r y 16, 1901, pp. 267-268. 3 GRAHAM, C . K.—^Poultry Observations. 1. Causes of Death of Young Chicks. Bulletin No. 44, November, 1906. Storrs A g r i c u l t u r a l Experim e n t Station. * RETTGER, L·. F., AND HARVEY, S . C . — F a t a l Septicaemia in Young Chickens, or "White D i a r r h e a . " J o u r n a l of Medical Research, Vol. 18, N. S. Vol. 13, No. 2, May, 1908, pp. 277-290. 5 M I L K S , H . J.—A P r e l i m i n a r y Report on Some Diseases of Chickens. Bull. No. 108, August, 1908. Louisiana Agricultural E x p e r i m e n t Station. β RETTGER, L . F . — F u r t h e r Studies on F a t a l Septicaemia in Young Chickens, or " W h i t e D i a r r h e a . " J o u r n a l of Medical Research, Vol. 21, No. 1 (N. S. Vol. 16, No. 1 ) , July, 1909, pp. 115-123. 7 RETTGER, L . F . , AND STONEBURN, F . H . — B a c i l l a r y W h i t e D i a r r h e a of Young Chicks. Bulletin No. 60, December, 1909. S t o r r s Agricultural E x p e r i m e n t Station^ s JONES, F . S.—Fatal S e p t i c e m i a or Bacillary W h i t e D i a r r h e a in Young Chickens. Report of t h e New York S t a t e V e t e r i n a r y College for 1910, pp. 111-129. 9 RETTGEE, L . F . , AND STONEBURN, F . H . — B a c i l l a r y W h i t e D i a r r h e a of Young Chicks. (Second R e p o r t ) Bulletin No. 68, April, 1911. S t o r r s Agricultural E x p e r i m e n t Station. 1» GAGE, G . E . — N o t e s on Ovarian Infection w i t h B a c t e r i u m P u l l o r u m ( R e t t g e r ) in t h e Domestic Fowl. J o u r n a l of Medical Research, Vol 24. (N. S. Vol. 19), No. 3, J u n e , 1911.
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appearance of agglutinins in the blood. Furthermore, such freed flocks may acquire infection from contaminated runs and houses after the infected fowls have been eliminated. Annual testing is therefore to be recommended until the flock is shown to be free on two consecutive tests at least. Acute infection with the white diarrhea organism may occur in adult fowls as a septicemia. Such infection may be exogenous or endogenous in origin. The disease may be contracted through the eating of eggs infected with the organism. Some fowls appear to recover from such infection, but they may retain the infection localized in the ovary or other parts of the body and thus be permanent carriers. Endogenous infection may arise from an infection of the ovary. In concluding, it might be said that we have full knowledge regarding the mode of transmission and control of this disease and it remains only a matter of applying this information.
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1* RBTTGEK, L·. F . , KiRKPATRiCK, W M . F . , AND STONEÎBURN, F . H.—Bacil-
lary White D i a r r h e a of Young Chicks. ( T h i r d R e p o r t ) Bulletin No. 74, December, 1912. S t o r r s A g r i c u l t u r a l E x p e r i m e n t Station. 15 BusHNBLi, L. D., AND MATJEIR, 0.—The Use of Milk Cultures of B. Bulgaricus in t h e Prevention and T r e a t m e n t of Bacillary W h i t e Diarrhea. A m e r i c a n V e t e r i n a r y Review, November, 1913, pp. 194-207. 10 GAGE, G . B . , PAIGE, B . H . , AND H Ï L A N D , H . W . — O n t h e Diagnosis of
Infection with B a c t e r i u m P u l l o r u m in t h e Domestic Fowl. Bulletin No. 148, April, 1914. Massachusetts A g r i c u l t u r a l E x p e r i m e n t Station. 17 RETTGER, L . F . — O v a r i a n Infection in t h e Domestic Fowl a n d Direct T r a n s m i s s i o n of Disease to the Offspring. J o u r n a l of E x p e r i m e n t a l Medicine, Vol. 19, No. 6, 1914, pp. 552-561. 18 RETTGER, L·. F., KIRKPATRICK, W M . F . , AND JONES, R . E . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks. A g r i c u l t u r a l E x p e r i m e n t Station.
Bulletin No. 77, J u n e , 1914, S t o r r s
19 RETTGER, L . F . , KIRKPATRICK, W M . P., AND CARD, L . E . — M i l k Feed-
ing a n d I t s Influence on Growth and Mortality. Comparative Study of t h e Value of Sweet and Sour Milk. Bulletin No. 80, April, 1915, S t o r r s Agricultural E x p e r i m e n t Station. 20 S M I T H , T H . , AND T E N BROKCK, CARL.—A Note on t h e R e l a t i o n be-
tween B. P u l l o r u m ( R e t t g e r ) and the Fowl Typhoid Bacillus J o u r n a l of Medical Research, Vol. 31, 1915, pp. 547-555. 21 GAGE, G . E . , AND PAIGE, B . H . — B a c i l l a r y W h i t e D i a r r h e a ium P u l l o r u m Infection) in Young Chicks in Massachusetts. No. 163, August, 1915. Massachusetts A g r i c u l t u r a l E x p e r i m e n t
(Moore), (BacterBulletin Station.
22 RETTGER, L . F . , KIRKPATRICK, W M . F . , AND J O N E S , R . B . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks, (Fifth R é p o r t ) Bulletin No. 85, December, 1915. S t o r r s A g r i c u l t u r a l E x p e r i m e n t Station. 23 GAGE, G . B . , AND MARTIN,-JAMES.^-Notes on t h e Histo-Pathology of t h e I n t e s t i n e s in Young Chicks Infected w i t h B a c t e r i u m Pullorum. J o u r n a l of Medical Research, Vol. 34, 1916, pp. 149-156. 2* RETTGER, L . F . , H i n x , T. G., AND STTJRGES, W M . S.—Feeding Experim e n t s with B a c t e r i u m P u l l o r u m . T h e Toxicity of Infected Eggs. Journal of E x p e r i m e n t a l Medicine, Vol. 23, No. 4, April, 1916, pp. 475-489. 25 RETTGER, L . F . , KIRKPATRICK, W M . F . , AND J O N E S , R . E . — B a c i l l a r y
W h i t e D i a r r h e a of Young Chicks. ( S i x t h R e p o r t ) Bulletin No. 88, Oc-
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