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Control of Infectious Synovitis
LEPTOSPIRA POMONA INFECTION SUMMARY
ACKNOWLEDGMENT
The author gratefully acknowledges the cooperation and guidance received from Drs. S. G. Kenzy and R. W. H. Gillespie. REFERENCES Bernkopf, H., 1948: Experimental leptospira infec-
tions in chickens. Proc. Soc. Expt. Biol. Med. 67: 148. Byrne, R. J., W. Jahnes, and C. A. Gloiser, 1955: Studies on experimental avian leptospirosis. Cornell Vet. 45:290-296. Gillespie, R. W. H., S. G. Kenzy, L. Ringen and F. K. Bracken, 1953. Studies on leptospirosis in cattle—outline, of progress. Stockman's Handbook, December, 1953, p. 76. Hoag, W. G., W. S. Gochenour, Jr. and R. H. Yager, 1953: Use of baby chicks for isolation of leptospires. Proc. Soc. Expt. Biol. Med. 83: 712-713. Kenzy, S. G., 1954. Unpublished data. State College of Washington, Pullman, Wash. Michin, and Azinow, 1937: Spirochaetal jaundice of cattle of North Caucasus. Abstract in Vet. Bull., 7: 419, original not seen. Ringen, L. M., and W. Okazaki, 1956. The susceptibility of several different laboratory animals to infection with L. pomona. J. Infect. Diseases. Rocky Mountain Laboratory, Leptospira Unit, 1952: Leptospirological techniques. Circ. Letter, 1952. Symposium on the Leptospiroses, 1952. Army Medical Service Graduate School, Walter Reed Army Medical Center.
Control of Infectious Synovitis 1. CONTINUOUS FEEDING OF ANTIBIOTICS AND THE INFLUENCE OF DIETHYLSTILBESTROL AND COCCIDIOSTATS 1 D. C. SHELTON, J. K. BLETNER, 2 N. 0. OLSON, G. C. ANDERSON AND C. E. WEAKLEY, JR. Departments of Agricultural Biochemistry and Animal Husbandry, West Virginia Agricultural Experiment Station, Morgantown (Received for publication July 16, 1956)
I
NFECTIOUS synovitis, caused by a filterable agent, was first described by Olson et al. (1954 a,b) and Wills (1954). Further descriptions of the disease have been made by Olson et al. (1955, 1956), Wills (1955), Wills and Delaplane (1955), Cover and Galeta (1955) and Cover et al. (1956). Snoeyenbos and Olesiuk (1955) reported a similar disease in turkeys. The occurrence of this disease in broiler and replacement flocks has been important
primarily in the eastern and southern states. Unlike many other diseases economic losses from mortality are secondary 1 Published with the approval of the Director of the West Virginia Agricultural Experiment Station as Scientific Paper No. 537. Support provided in part by funds allocated to regional project NE-5 C9b3) and by funds from Abbott Laboratories, North Chicago, Illinois. 2 Present address, Department of Poultry Science, Ohio State University, Columbus.
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1. The feeding of diluted urine from a heifer with leptospiruria produced a positive agglutination-lysis test for Leptospira pomona in four of six White Leghorn cockerels; two of three Hungarian partridges; one of eight Pekin ducks; and two of two Ring-necked pheasants. 2. An attempt to demonstrate the transmission of leptospirae in the excreta of birds positive to the agglutinationlysis test for Leptospira pomona by inoculating diluted fecal samples intraperitoneally into two-day-old chicks was unsuccessful.
113
114
SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
EXPERIMENTAL
TABLE 1.—Basal ration
Ground yellow corn Soybean oil meal (50% protein) Mineral mixture 1 Vitamin mixture 2
Number 1 2 3 4 5
6 7
Day old New Hampshire chicks obtained from the University flock were placed in electrically heated batteries equipped with raised wire floors, Twelve unsexed chicks were selected at random, wingbanded and placed in each battery. During the first week all chicks were fed the basal ration described in Table 1. At the end of this time the chicks were divided into four major groups. One group was continued on the basal ration whereas the others received the basal ration containing either crystalline aureomycin (chlortetracycline hydrochloride), terramycin (oxytetracycline hydrochloride) or erythromycin at a level of 200 grams per ton of feed. Each major group was further divided into seven sub-groups
Ingredient
TABLE 2.—Experimental treatment within each major group
Percent 64.0 31.0 5.0
1 Mineral mixture was composed of: 50% dicalcium phosphate, 30% limestone and 20% iodized salt. 2 Vitamin mixture per 100 lbs. of diet: 22.7 gm. dry vitamin A (10,000 IU/gm., 2.27 gm. dry vitamin D 3 (15,000 IU/gm. ( 150 mg. riboflavin, 750 meg. vitamin B12 and 36 mg. vitamin K (menadione).
Additional Treatment (12 birds per treatment) None (Basal Ration only). Diethylstilbestrol pellet (12 mg.) implanted at the base of skull following inoculation Sulfaquinoxaline 0.0175 percent in the feed—fed continuously from one week of age Treatments 2 plus 3 Sulfaquinoxaline 0.04 percent in drinking water—began on fifth day following inoculation and given on an intermittent schedule as commonly recommended for treatment of coccidiosis Treatment 5 plus nitrophenide 0.025 percent in the feed—fed continuously from one week of age Sulfamethazine 1:1,000 in the drinking water—began on fifth day following inoculation and given on an intermittent schedule as commonly recommended for treatment of coccidiosis
each of which was assigned to one of the treatments described in Table 2. Blood studies were initiated when the birds became three weeks of age. A suitable sample of blood was obtained from a wing vein puncture. Hemoglobin determinations were made by the acid hematin method. Blood smears for differential white cell counts were made and stained with a combination of MayGruenwald and Giemsa stains. After blood samples were obtained, six birds in each battery were randomly selected and inoculated with a virulent culture of the infectious synovitis agent. The material used was a joint exudate suspension of three chicken grown isolates which have been described by Olson et al. (1956) and designated as 36H, 29H and 29M. Equal portions of these cultures were carefully mixed and stored at — 20°C. Immediately prior to usage they were thawed and 0.25 ml. portions were injected into the synovial membranes of the left footpad. To follow the progress of the infection
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to those from morbidity. Affected birds become unmarketable because of the infected joints and emaciation which occurs in the later stages of the infection. Preliminary reports on the value of some antibiotics used as preventive and control measures have been presented by Bletner et al. (1955), Olson et al. (1955) and Shelton et al. (1955). This report describes in greater detail the therapeutic value of aureomycin, terramycin and erythromycin and the influence of stilbestrol and certain coccidiostats on the control of infectious synovitis.
115
CONTROL OF INFECTIOUS SYNOVITIS
RESULTS AND DISCUSSION Influence of diethylstilbestrol, sulfaquinoxaline, nitrophenide and sulfametha-
zine. The use of diethylstilbestrol, sulfaquinoxaline, nitrophenide, sulfamethazine or combinations of these drugs as given in Table 2 had no detrimental or therapeutic influence on the course of the disease. However, it is not known what effect these materials might have exerted if the disease had been established by contact infection. A few birds receiving sulfaquinoxaline in the drinking water, treatments 5 and 6, exhibited areas of subcutaneous hemorrhages. Whole blood clotting times made from birds on these two treatments were markedly elevated. Changes in hemoglobin values and differential white cell counts. Olson et al. (1956) have observed that infectious synovitis caused a reduced erythocyte count and hemoglobin level. However, the leucocyte count was increased. The results of a controlled study to determine the changes in hemoglobin level and differential leucocycte count is found in Table 3. In eleven days the synovitis infection in the control inoculated birds had reduced the hemoglobin level to an average of 7.3 gm. per 100 ml. blood. On the other hand, contact controls appeared normal and maintained a normal hemo-
T A B I E 3.—Influence of infectious synovitis on hemoglobin level and differential leucocyte count of chicks with and without therapeutic levels of antibiotics No. of birds
Hb, gms./lOO ml.
Control Contact Inoculated
41 42
Erythromycin Contact Inoculated
Treatment
Differential white cell count Lymph
Het.
Mono.
Eos.
Baso.
Im.
9.0 7.3
66 38
24 39
9.0 16.0
.6 .4
.2 .1
1.1 6.1
41 42
9.3 7.0
66 34
23 41
10.3 18.7
.4 .1
.3 .1
.3 6.7
Aureomycin Contact Inoculated
41 41
9.0 9.0
69 67
23 25
8.1 7.7
.3 .4
.2 .1
.1 .1
Terramycin Contact Inoculated
41 42
9.1 9.0
67 67
24 24
8.7 8.9
.2 .2
.1 .1
.1 .1
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the birds were observed carefully each day. In addition all birds were individually examined and weighed twice weekly. Swellings characteristic of infectious synovitis were evident in the inoculated birds by the fourth day. On the next day, the fifth following inoculation, three batteries of birds which had previously received the basal ration were given rations containing 200 grams per ton of aureomycin, terramycin or erythromycin. Blood studies conducted earlier were repeated eleven days after inoculation with the infectious synovitis agent. At this time the infected birds were showing severe external lesions and considerable morbidity although very few deaths had occurred. The experiment was continued until 25 days had elapsed following inoculation at which time the birds were seven weeks old. Some birds from selected treatments were retained for further study. All other birds were killed and examined for internal pathology.
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SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
Body Weight Gains. Even though unsexed chicks were used in this experiment it is evident from Figure 1 that the infection greatly reduced weight gains. The top growth curves represent the growth response for a 25-day period of the uninoculated and the inoculated birds fed the
21 23
30
34
37
AGE [PAYS -2
0
4
7
II
14
18
DAYS AFTER INOCULATION
FIG. 1. Influence of infectious synovitis on the growth of chicks. Note the normal growth of infected chicks when aureomycin or terramycin were fed continuously.
basal ration. At first the infected birds continued to grow at a fairly normal rate. Then the growth rate slowed until the birds just maintained themselves. After the infection became severe and impaired the locomotion of the birds, dehydration occurred and weight losses became evident. Effect of the Antibiotics. Results presented in Table 3 and Figure 1 showed that under conditions of this experiment crystalline erythromycin was ineffective against this synovitis agent. However, when crystalline aureomycin or terramycin was included in the feed at 200 grams per ton, both were effective in preventing signs of synovitis. In these birds medication was started 16 days before inoculation and given continuously thereafter. The weight gains, hemoglobin values and differential white cell counts appeared to be the same for the contact controls and the experimentally infected birds. If the administration of aureomycin or terramycin was delayed until swellings occurred in the inoculated footpad, remission of the symptoms differed. When aureomycin therapy (200 grams per ton) was provided, all visible swellings disappeared within 10 days. The growth rate (Figure 2), hemoglobin level and differential white cell count (Table 4) appeared to be the same in the aureomycin-treated inoculated and the uninoculated birds. Similar delayed treatment with terramycin did not seem to cause regression of the swellings in the infected birds. However, the swellings seemed to be localized in the inoculated footpad. Although the growth rate (Figure 2) of these terramycin-treated birds was essentially the same as for the non-inoculated birds, there were slight differences observed in hematology studies (Table 4). Lower hemoglobin values, a reduction in the
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globin level of 9.0 gm. per 100 ml. blood. Likewise, there were marked changes in the differential white cell count. The percentage of lymphocytes was reduced substantially. The inoculated controls had an average of 38 percent as compared to 66 percent lymphocytes for the contact controls. As the lymphocytes decreased, an increase in the percentage of heterophils, monocytes and immature forms occurred concomitantly. In the inoculated controls the average counts for the heterophils, monocytes and immature forms were 39, 16 and 6.1 percent respectively whereas the contact controls had average counts of 24, 9 and 1.1 percent respectively.
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CONTROL or INFECTIOUS SYNOVITIS TABLE 4.—The influence of delayed antibiotic feeding on hemoglobin level and differential white cell counts No. of birds
Hb, gms./lOO ml.
Control Contact Infected
6 4
Erythromycin Contact Infected
Treatment
Differential white cell count Het.
Mono.
Eos.
Baso.
Im.
9.0 7.3
59 31
27 45
12.3 13.5
.7 .8
0 0
1.8 6.8
6 6
9.6 8.0
68 39
22 40
8.8 16.0
.3 .2
0 .2
1.0 5.2
Aureomycin Contact Infected
6 6
9.5 9.4
77 75
17 16
6.2 7.8
.3 .4
0 0
0 0
Terramycin Contact Infected
6 6
9.4 8.2
69 59
22 26
9.2 15.0
.2 0
0 0
0
percentage of lymphocytes and an increase in the percentage of monocytes were observed and suggested that terramycin did not completely control the infection in these birds. The data presented in Tables 3 and 4 covers the development of the disease during the first eleven days. The number
30
21 23 -2
O
AGE, 4
7
34 37 DAYS II 14
DAYS AFTIR
18
INOCULATION
FIG. 2. Influence on the growth of infected chicks when the feeding of aureomycin and terramycin was delayed till signs of infectious synovitis were visible.
.7
of infected chicks, body weights and mortality for the 25-day period are presented in Table 5. Nearly every bird which had been experimentally infected developed a slight swelling in the inoculated footpad by the fourth day following inoculation. By the seventh day the swellings in the birds which had been receiving aureomycin or terramycin disappeared. However, the inoculated control and erythromycin-treated groups became severely infected as shown by the slow rate of gain and the high mortality. Examination of the internal organs at necropsy was made on all control and erythromycin-treated birds and on half of the aureomycin- and terramycin-treated birds. Internal Lesions. Classifications of the condition of those organs which appeared to be abnormal are given in Table 6. The spleen is frequently involved. In the early stages of the disease, enlargement is commonly seen. Later, the spleen may appear smaller than normal. In this study it would appear that splenomegaly was not a consistent lesion. The incidence of splenomegaly among the contact controls or contact erythromycin-treated
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Lymph
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SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
TABLE 5.—Incidence of lesions, body weights and •mortality 25 days after experimental infection
Treatment
MorNo. Weight, tality due to with gms. synolesions vitis
No. of birds
41 42
0 41
805 619
0 24
Erythromycin 1 Contact Inoculated
42 42
0 42
824 596
0 32
Aureomycin1 Contact Inoculated
41 41
0 0
850 815
0 0
Terramycin 1 Contact Inoculated
41 42
0 0
806 811
0 0
1 Continuous administration of antibiotics began 16 days prior to infection.
birds may indicate infection through contact. However, no joint involvement or other identifiable signs were present, and therefore, verification of infection by contact was not established. To examine the bone marrow, both the femur and tibia were cut at their centers. A normal appearing bone marrow was considered to be one which showed a pale center surrounded by a red periphery. This indicated that the marrow in the red periphery was more active than that in
TABLE 6.—Summary of observations made on internal organs Spleen
Bone marrow
Treatment
No. of birds
Small
Control Contact Inoculated
42 42
8 5
24 22
10 IS
Erythromycin Contact Inoculated
48 48
2 6
36 27
Aureomycin Contact Inoculated
24 23
5 6
Terramycin Contact Inoculated
23 22
4 3
1 2
Pale center, red periphery. Marrow red throughout.
Liver
Kidney Normal
Enlarged
Discolored
Reds
Normal
Enlarged
35 26
7 16
42 21
0 21
42 22
0 13
0 17
10 15
45 27
3 21
47 21
1 28
42 22
0 13
0 IS
17 14
2 3
20 18
4 5
24 23
0 0
24 22
0 1
0 0
19 19
0 0
18 22
5 0
22 22
1 0
23 22
0 0
0 0
EnNormal larged
Normal
1
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Control Contact Inoculated
the pale center and under normal conditions would supply sufficient quantities of erythrocytes. As indicated in Table 6, the marrow observed in some of the infected birds was red throughout. Since in affected birds the hemoglobin content is reduced as well as the number of red cells, the demand for erythrocytes is great. As a result, the area of erythropoiesis was greatly extended and appeared to involve the entire marrow. In approximately 50 percent of the infected birds the kidneys were enlarged and often pale. Enlarged and/or discolored livers among the infected birds were observed frequently. In advanced cases a greenish discoloration of the liver was more common than in less severe cases. In general observations on the internal organs as a means of diagnosis can be very misleading. The stage of the infection has much to do with the pathological changes of the organs. This is illustrated by the splenomegaly which was more common in the early stages, whereas kidney and liver enlargement were more frequent to the advanced stages. Duration of Antibiotic Treatment. Preliminary information concerning the length of time which antibiotics must be
CONTROL OF INFECTIOUS SYNOVITIS
119
TABLE 7.—Occurrence of infectious synovitis as influenced by antibiotic treatment Treatment Group No. 1 2 3 4 5 6
Controls—not previously exposed to infection Aureomycin—continuous Terramycin—continuous Aureomycin—discontinued at 7 weeks Terramycin—discontinued at 7 weeks Aureomycin—started after signs—discontinued at 7 weeks Terramycin—started after signs—discontinued at 7 weeks
fed to experimentally infected birds to prevent the infection from occurring was obtained by continued observations on certain birds used to obtain the data presented above. At this time all birds were seven weeks old and as shown in Table 7, only two birds had signs of infection. Those birds were in group 7 in which terramycin had been provided after signs of synovitis had appeared. Birds in group 1 had received a commercial ration and had not previously been exposed by contact or experimental infection. The previous and subsequent treatment of groups 2 through 7 are indicated in Table 7. For example, group 2 had received 200 grams of aureomycin per ton of feed since they were a week old. This treatment was continued for the additional period of three weeks at which time the birds were 10 weeks old. For these groups, birds were selected so as to obtain equal numbers of those which had been previously inoculated at 23 days of age and those which had not been inoculated. Reoccurrence of signs of infectious synovitis was apparent in groups 5 to 7. Continuous feeding of aureomycin and terramycin was effective in preventing development of the condition as shown in groups 2 and 3. The continuous feeding
No. birds
Age, weeks 7
8
9
10
6 12 12 12 12
0 0 0 0 0
0 0 0 0 1
0 0 0 0 1
0 0 0 0 2
6
0
2
2
2
6
2
3
3
4
of aureomycin for six weeks and discontinued at seven weeks, group 4, prevented the reoccurrence of any signs of the disease. Similar treatment with terramycin, group 5, did not afford the same protection. In this group two birds developed the disease. One was previously inoculated and the other was a contact control. In groups 6 and 7 where signs of synovitis were evident before the antibiotics were given, the removal of either drug resulted in a reoccurrence of the infection. Signs reappeared within a week in two of the three birds of group 6 which had been previously inoculated. In the terramycin-treated group all three previously inoculated birds and one of three contact controls became infected. The incubation period of infectious synovitis in contact control chickens has been approximately 30 days or longer (Olson et al., 1956). It appeared that the contact infection must have occurred prior to the time when the terramycin feeding was discontinued. From the results obtained it seems evident that neither aureomycin nor terramycin completely destroyed the infectious synovitis agent once a build-up of the disease had taken place. Aureomycin
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7
Description
Occurrence of lissions
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SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
treatment brings about a regression of signs if treatment is started immediately upon their appearance. Terramycin was less effective. SUMMARY
ACKNOWLEDGEMENT
The authors are indebted to the following concerns for material gratuitously supplied during the course of this investigation: Abbott Laboratories, North Chicago, Illinois for crystalline erythromycin; Chas. Pfizer & Co., Inc., Terre Haute, Indiana for crystalline terramycin hydrochloride; Lederle Laboratories Division, American Cyanamid Company,
REFERENCES Bletner, J. K., D. C. Shelton, N. O. Olson and C. E. Weakley, Jr., 1955. Synovitis control. 3. The efficacy of aureomycin with relation to time of inoculation. Poultry Sci. 34:1181. Cover, M. S., and J. N. Galeta, 1955. Studies on avian arthritis of chickens. Proceedings 27th Annual Pullorum Conference, University of New Hampshire, Durham, New Hampshire. Cover, M. S., J. N. Galeta and E. F. Waller, 1956. The etiology of an arthritic disease of chickens. Am. J. Vet. Res. 17: 12-15. Olson, N. O., J. K. Bletner, D. C. Shelton, D. A. Munro and G. C. Anderson, 1954a. Enlarged joint condition in poultry caused by an infectious agent. Proceedings of West Virginia Feed Dealer's Short Course, April 19,1954. Olson, N. O., J. K. Bletner, D. C. Shelton, D. A. Munro and G. C. Anderson, 1954b. Enlarged joint condition in poultry caused by an infectious -•• agent. Poultry Sci. 33:1075. Olson, N. O., J. K. Bletner, D. C. Shelton, D. A. Munro and G. C. Anderson, 1955. Infectious anemia-synovitis. Science Serves Your Farm, Annual Report published quarterly, Agric. Exp. Sta., West Virginia University. Olson, N. O., D. C. Shelton, J. K. Bletner, D. A. Munro and G. C. Anderson, 1956. Studies of infectious synovitis in chickens. Am. J. Vet. Res. 17:747. Olson, N. O., D. C. Shelton, J. K. Bletner and C. E. Weakley, Jr., 1955. Synovitis control. 2. A comparison of levels of antibiotics. Poultry Sci. 34: 1214. Shelton, D. C , J. K. Bletner, N. O. Olson, G. C. Anderson and C. E. Weakley, Jr., 1955. Synovitis control. 1. Continuous feeding of antibiotics and the influence of stilbestrol and coccidiostats. Poultry Sci. 34: 1221. Snoeyenbos, G. H., and O. M. Olesiuk, 1955. Studies of an agent producing arthritis in turkeys. Proceedings 27th Annual Pullorum Conference, University of New Hampshire, Durham, N. H. Wills, F. K., 1954. Preliminary report on transmission of an agent producing an arthritis in chick-
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The use of crystalline aureomycin, terramycin and erythromycin in the control of infectious synovitis has been investigated under laboratory conditions. Erythromycin was ineffective against the infectious agent used. However, 200 grams of aureomycin or terramycin per ton of feed was effective when fed continuously. Terramycin appeared less effective when treatment was delayed until after the birds showed signs of synovitis. Hemoglobin values were lowered substantially in infected birds. There were marked changes in the differential white cell counts. Infected birds showed a reduced percentage of lymphocytes, but an increased percentage of heterophils, monocytes and immature forms of leucocytes. Observations were made on internal organs. The pathological changes of these organs are influenced by the stage of the infection. The use of diethylstilbestrol, sulfaquinoxaline, nitrophenide, sulfamethazine or combinations of these drugs had no detrimental or therapeutic influence on the course of the disease.
Pearl River, New York for crystalline aureomycin hydrochloride; Merck and Co., Inc., Rahway, New Jersey for Bcomplex vitamins and menadione; and Nopco Chemical Company, Harrison, New Jersey for vitamin A (Nopcay 10) and vitamin D 3 (Super Nopdex 15).
CONTROL OF INFECTIOUS SYNOVITIS ens. Progress Report 1674. Texas Agric. Exp. Sta., College Station, Texas, April 26,1954. Wills, F. K., 1955. Studies of an unidentified agent producing arthritis in chickens. Southwestern Vet. 8: Part 2.
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Wills, F. K., and J. P. Delaplane, 1955. Transmission and therapy on an agent which produces arthritis in chickens. Proc. 92nd meeting, Am. Vet. Med. Assoc. 350-356.
The Efficacy of Chlortetracycline at Several Temperatures in Controlling Spoilage of Poultry Meat* W. G. SHANNON1 AND W. J. STADELMAN (Received for publication July 16, 1956) INTRODUCTION
T
HE volume of poultry meat moving through market channels is increasing each year. A high percentage of this meat is fryer chickens most of which are delivered to the final consumer without freezing. One of the major problems in handling the fresh fryers is the short keeping time. The spoilage of fresh poultry meat held under refrigeration without freezing is predominantly bacterial. This information has been available for years. Lockhead and Landerkin (1935) reported that spoilage of chicken held at refrigerator temperatures above freezing was due primarily to surface bacteria. Further confirmation comes from the work of Ayres, Ogilvy and Stewart (1950). The appearance of slime nodules and off-odor has been commonly used to determine spoilage, but Ziegler, Spencer and Stadelman (1954) report a rapid microscopic method for this determination. This was the method used in this study.
On November 30, 1955, the Federal Food and Drug Administration cleared chlortetracycline for use "in and on uncooked poultry." Chlortetracycline is * Journal Paper No. 1001 of the Purdue University Agricultural Experiment Station. 1 Present Address: Loughry Agricultural College, Cookstown County, Tyrone, North Ireland.
recognized as one of the most effective of the broad spectrum antibiotics and within recent years a vast amount of work has been done in both the fields of human medicine and agriculture on the use of chlortetracycline. Ziegler and Stadelman (1955) report on the effectiveness of chlortetracycline in the retardation of spoilage of poultry meat while Kohler, Miller and Broquist (1955) report chlortetracycline being the most effective of several antibiotics which they used. The purpose of this paper is to report on the effect of chlortetracycline treatment of eviscerated poultry held at different levels of temperature. EXPERIMENTAL PROCEDURE
The study consisted of two treatments at five temperature levels of storage. It was conducted in two equal parts. In the first the three storage temperatures used were 32°, 37°, and 42°F. and in the'second 42°, 47°, and 68°F. There were two replicates at each temperature and six birds in each replicate. The birds used were fryers which were dressed using a batch scalder at 140°F. for 40 seconds. After picking on a mechanical picker they were cooled in ice water, eviscerated and then further cooled in ice. Each bird was divided into halves, one half being used in treatment 1, the other in treatment 2.
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Purdue University, Agricultural Experiment Station, Lafayette, Indiana
ACKNOWLEDGMENT
The author gratefully acknowledges the cooperation and guidance received from Drs. S. G. Kenzy and R. W. H. Gillespie. REFERENCES Bernkopf, H., 1948: Experimental leptospira infec-
tions in chickens. Proc. Soc. Expt. Biol. Med. 67: 148. Byrne, R. J., W. Jahnes, and C. A. Gloiser, 1955: Studies on experimental avian leptospirosis. Cornell Vet. 45:290-296. Gillespie, R. W. H., S. G. Kenzy, L. Ringen and F. K. Bracken, 1953. Studies on leptospirosis in cattle—outline, of progress. Stockman's Handbook, December, 1953, p. 76. Hoag, W. G., W. S. Gochenour, Jr. and R. H. Yager, 1953: Use of baby chicks for isolation of leptospires. Proc. Soc. Expt. Biol. Med. 83: 712-713. Kenzy, S. G., 1954. Unpublished data. State College of Washington, Pullman, Wash. Michin, and Azinow, 1937: Spirochaetal jaundice of cattle of North Caucasus. Abstract in Vet. Bull., 7: 419, original not seen. Ringen, L. M., and W. Okazaki, 1956. The susceptibility of several different laboratory animals to infection with L. pomona. J. Infect. Diseases. Rocky Mountain Laboratory, Leptospira Unit, 1952: Leptospirological techniques. Circ. Letter, 1952. Symposium on the Leptospiroses, 1952. Army Medical Service Graduate School, Walter Reed Army Medical Center.
Control of Infectious Synovitis 1. CONTINUOUS FEEDING OF ANTIBIOTICS AND THE INFLUENCE OF DIETHYLSTILBESTROL AND COCCIDIOSTATS 1 D. C. SHELTON, J. K. BLETNER, 2 N. 0. OLSON, G. C. ANDERSON AND C. E. WEAKLEY, JR. Departments of Agricultural Biochemistry and Animal Husbandry, West Virginia Agricultural Experiment Station, Morgantown (Received for publication July 16, 1956)
I
NFECTIOUS synovitis, caused by a filterable agent, was first described by Olson et al. (1954 a,b) and Wills (1954). Further descriptions of the disease have been made by Olson et al. (1955, 1956), Wills (1955), Wills and Delaplane (1955), Cover and Galeta (1955) and Cover et al. (1956). Snoeyenbos and Olesiuk (1955) reported a similar disease in turkeys. The occurrence of this disease in broiler and replacement flocks has been important
primarily in the eastern and southern states. Unlike many other diseases economic losses from mortality are secondary 1 Published with the approval of the Director of the West Virginia Agricultural Experiment Station as Scientific Paper No. 537. Support provided in part by funds allocated to regional project NE-5 C9b3) and by funds from Abbott Laboratories, North Chicago, Illinois. 2 Present address, Department of Poultry Science, Ohio State University, Columbus.
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1. The feeding of diluted urine from a heifer with leptospiruria produced a positive agglutination-lysis test for Leptospira pomona in four of six White Leghorn cockerels; two of three Hungarian partridges; one of eight Pekin ducks; and two of two Ring-necked pheasants. 2. An attempt to demonstrate the transmission of leptospirae in the excreta of birds positive to the agglutinationlysis test for Leptospira pomona by inoculating diluted fecal samples intraperitoneally into two-day-old chicks was unsuccessful.
113
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SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
EXPERIMENTAL
TABLE 1.—Basal ration
Ground yellow corn Soybean oil meal (50% protein) Mineral mixture 1 Vitamin mixture 2
Number 1 2 3 4 5
6 7
Day old New Hampshire chicks obtained from the University flock were placed in electrically heated batteries equipped with raised wire floors, Twelve unsexed chicks were selected at random, wingbanded and placed in each battery. During the first week all chicks were fed the basal ration described in Table 1. At the end of this time the chicks were divided into four major groups. One group was continued on the basal ration whereas the others received the basal ration containing either crystalline aureomycin (chlortetracycline hydrochloride), terramycin (oxytetracycline hydrochloride) or erythromycin at a level of 200 grams per ton of feed. Each major group was further divided into seven sub-groups
Ingredient
TABLE 2.—Experimental treatment within each major group
Percent 64.0 31.0 5.0
1 Mineral mixture was composed of: 50% dicalcium phosphate, 30% limestone and 20% iodized salt. 2 Vitamin mixture per 100 lbs. of diet: 22.7 gm. dry vitamin A (10,000 IU/gm., 2.27 gm. dry vitamin D 3 (15,000 IU/gm. ( 150 mg. riboflavin, 750 meg. vitamin B12 and 36 mg. vitamin K (menadione).
Additional Treatment (12 birds per treatment) None (Basal Ration only). Diethylstilbestrol pellet (12 mg.) implanted at the base of skull following inoculation Sulfaquinoxaline 0.0175 percent in the feed—fed continuously from one week of age Treatments 2 plus 3 Sulfaquinoxaline 0.04 percent in drinking water—began on fifth day following inoculation and given on an intermittent schedule as commonly recommended for treatment of coccidiosis Treatment 5 plus nitrophenide 0.025 percent in the feed—fed continuously from one week of age Sulfamethazine 1:1,000 in the drinking water—began on fifth day following inoculation and given on an intermittent schedule as commonly recommended for treatment of coccidiosis
each of which was assigned to one of the treatments described in Table 2. Blood studies were initiated when the birds became three weeks of age. A suitable sample of blood was obtained from a wing vein puncture. Hemoglobin determinations were made by the acid hematin method. Blood smears for differential white cell counts were made and stained with a combination of MayGruenwald and Giemsa stains. After blood samples were obtained, six birds in each battery were randomly selected and inoculated with a virulent culture of the infectious synovitis agent. The material used was a joint exudate suspension of three chicken grown isolates which have been described by Olson et al. (1956) and designated as 36H, 29H and 29M. Equal portions of these cultures were carefully mixed and stored at — 20°C. Immediately prior to usage they were thawed and 0.25 ml. portions were injected into the synovial membranes of the left footpad. To follow the progress of the infection
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to those from morbidity. Affected birds become unmarketable because of the infected joints and emaciation which occurs in the later stages of the infection. Preliminary reports on the value of some antibiotics used as preventive and control measures have been presented by Bletner et al. (1955), Olson et al. (1955) and Shelton et al. (1955). This report describes in greater detail the therapeutic value of aureomycin, terramycin and erythromycin and the influence of stilbestrol and certain coccidiostats on the control of infectious synovitis.
115
CONTROL OF INFECTIOUS SYNOVITIS
RESULTS AND DISCUSSION Influence of diethylstilbestrol, sulfaquinoxaline, nitrophenide and sulfametha-
zine. The use of diethylstilbestrol, sulfaquinoxaline, nitrophenide, sulfamethazine or combinations of these drugs as given in Table 2 had no detrimental or therapeutic influence on the course of the disease. However, it is not known what effect these materials might have exerted if the disease had been established by contact infection. A few birds receiving sulfaquinoxaline in the drinking water, treatments 5 and 6, exhibited areas of subcutaneous hemorrhages. Whole blood clotting times made from birds on these two treatments were markedly elevated. Changes in hemoglobin values and differential white cell counts. Olson et al. (1956) have observed that infectious synovitis caused a reduced erythocyte count and hemoglobin level. However, the leucocyte count was increased. The results of a controlled study to determine the changes in hemoglobin level and differential leucocycte count is found in Table 3. In eleven days the synovitis infection in the control inoculated birds had reduced the hemoglobin level to an average of 7.3 gm. per 100 ml. blood. On the other hand, contact controls appeared normal and maintained a normal hemo-
T A B I E 3.—Influence of infectious synovitis on hemoglobin level and differential leucocyte count of chicks with and without therapeutic levels of antibiotics No. of birds
Hb, gms./lOO ml.
Control Contact Inoculated
41 42
Erythromycin Contact Inoculated
Treatment
Differential white cell count Lymph
Het.
Mono.
Eos.
Baso.
Im.
9.0 7.3
66 38
24 39
9.0 16.0
.6 .4
.2 .1
1.1 6.1
41 42
9.3 7.0
66 34
23 41
10.3 18.7
.4 .1
.3 .1
.3 6.7
Aureomycin Contact Inoculated
41 41
9.0 9.0
69 67
23 25
8.1 7.7
.3 .4
.2 .1
.1 .1
Terramycin Contact Inoculated
41 42
9.1 9.0
67 67
24 24
8.7 8.9
.2 .2
.1 .1
.1 .1
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the birds were observed carefully each day. In addition all birds were individually examined and weighed twice weekly. Swellings characteristic of infectious synovitis were evident in the inoculated birds by the fourth day. On the next day, the fifth following inoculation, three batteries of birds which had previously received the basal ration were given rations containing 200 grams per ton of aureomycin, terramycin or erythromycin. Blood studies conducted earlier were repeated eleven days after inoculation with the infectious synovitis agent. At this time the infected birds were showing severe external lesions and considerable morbidity although very few deaths had occurred. The experiment was continued until 25 days had elapsed following inoculation at which time the birds were seven weeks old. Some birds from selected treatments were retained for further study. All other birds were killed and examined for internal pathology.
116
SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
Body Weight Gains. Even though unsexed chicks were used in this experiment it is evident from Figure 1 that the infection greatly reduced weight gains. The top growth curves represent the growth response for a 25-day period of the uninoculated and the inoculated birds fed the
21 23
30
34
37
AGE [PAYS -2
0
4
7
II
14
18
DAYS AFTER INOCULATION
FIG. 1. Influence of infectious synovitis on the growth of chicks. Note the normal growth of infected chicks when aureomycin or terramycin were fed continuously.
basal ration. At first the infected birds continued to grow at a fairly normal rate. Then the growth rate slowed until the birds just maintained themselves. After the infection became severe and impaired the locomotion of the birds, dehydration occurred and weight losses became evident. Effect of the Antibiotics. Results presented in Table 3 and Figure 1 showed that under conditions of this experiment crystalline erythromycin was ineffective against this synovitis agent. However, when crystalline aureomycin or terramycin was included in the feed at 200 grams per ton, both were effective in preventing signs of synovitis. In these birds medication was started 16 days before inoculation and given continuously thereafter. The weight gains, hemoglobin values and differential white cell counts appeared to be the same for the contact controls and the experimentally infected birds. If the administration of aureomycin or terramycin was delayed until swellings occurred in the inoculated footpad, remission of the symptoms differed. When aureomycin therapy (200 grams per ton) was provided, all visible swellings disappeared within 10 days. The growth rate (Figure 2), hemoglobin level and differential white cell count (Table 4) appeared to be the same in the aureomycin-treated inoculated and the uninoculated birds. Similar delayed treatment with terramycin did not seem to cause regression of the swellings in the infected birds. However, the swellings seemed to be localized in the inoculated footpad. Although the growth rate (Figure 2) of these terramycin-treated birds was essentially the same as for the non-inoculated birds, there were slight differences observed in hematology studies (Table 4). Lower hemoglobin values, a reduction in the
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globin level of 9.0 gm. per 100 ml. blood. Likewise, there were marked changes in the differential white cell count. The percentage of lymphocytes was reduced substantially. The inoculated controls had an average of 38 percent as compared to 66 percent lymphocytes for the contact controls. As the lymphocytes decreased, an increase in the percentage of heterophils, monocytes and immature forms occurred concomitantly. In the inoculated controls the average counts for the heterophils, monocytes and immature forms were 39, 16 and 6.1 percent respectively whereas the contact controls had average counts of 24, 9 and 1.1 percent respectively.
117
CONTROL or INFECTIOUS SYNOVITIS TABLE 4.—The influence of delayed antibiotic feeding on hemoglobin level and differential white cell counts No. of birds
Hb, gms./lOO ml.
Control Contact Infected
6 4
Erythromycin Contact Infected
Treatment
Differential white cell count Het.
Mono.
Eos.
Baso.
Im.
9.0 7.3
59 31
27 45
12.3 13.5
.7 .8
0 0
1.8 6.8
6 6
9.6 8.0
68 39
22 40
8.8 16.0
.3 .2
0 .2
1.0 5.2
Aureomycin Contact Infected
6 6
9.5 9.4
77 75
17 16
6.2 7.8
.3 .4
0 0
0 0
Terramycin Contact Infected
6 6
9.4 8.2
69 59
22 26
9.2 15.0
.2 0
0 0
0
percentage of lymphocytes and an increase in the percentage of monocytes were observed and suggested that terramycin did not completely control the infection in these birds. The data presented in Tables 3 and 4 covers the development of the disease during the first eleven days. The number
30
21 23 -2
O
AGE, 4
7
34 37 DAYS II 14
DAYS AFTIR
18
INOCULATION
FIG. 2. Influence on the growth of infected chicks when the feeding of aureomycin and terramycin was delayed till signs of infectious synovitis were visible.
.7
of infected chicks, body weights and mortality for the 25-day period are presented in Table 5. Nearly every bird which had been experimentally infected developed a slight swelling in the inoculated footpad by the fourth day following inoculation. By the seventh day the swellings in the birds which had been receiving aureomycin or terramycin disappeared. However, the inoculated control and erythromycin-treated groups became severely infected as shown by the slow rate of gain and the high mortality. Examination of the internal organs at necropsy was made on all control and erythromycin-treated birds and on half of the aureomycin- and terramycin-treated birds. Internal Lesions. Classifications of the condition of those organs which appeared to be abnormal are given in Table 6. The spleen is frequently involved. In the early stages of the disease, enlargement is commonly seen. Later, the spleen may appear smaller than normal. In this study it would appear that splenomegaly was not a consistent lesion. The incidence of splenomegaly among the contact controls or contact erythromycin-treated
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Lymph
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SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
TABLE 5.—Incidence of lesions, body weights and •mortality 25 days after experimental infection
Treatment
MorNo. Weight, tality due to with gms. synolesions vitis
No. of birds
41 42
0 41
805 619
0 24
Erythromycin 1 Contact Inoculated
42 42
0 42
824 596
0 32
Aureomycin1 Contact Inoculated
41 41
0 0
850 815
0 0
Terramycin 1 Contact Inoculated
41 42
0 0
806 811
0 0
1 Continuous administration of antibiotics began 16 days prior to infection.
birds may indicate infection through contact. However, no joint involvement or other identifiable signs were present, and therefore, verification of infection by contact was not established. To examine the bone marrow, both the femur and tibia were cut at their centers. A normal appearing bone marrow was considered to be one which showed a pale center surrounded by a red periphery. This indicated that the marrow in the red periphery was more active than that in
TABLE 6.—Summary of observations made on internal organs Spleen
Bone marrow
Treatment
No. of birds
Small
Control Contact Inoculated
42 42
8 5
24 22
10 IS
Erythromycin Contact Inoculated
48 48
2 6
36 27
Aureomycin Contact Inoculated
24 23
5 6
Terramycin Contact Inoculated
23 22
4 3
1 2
Pale center, red periphery. Marrow red throughout.
Liver
Kidney Normal
Enlarged
Discolored
Reds
Normal
Enlarged
35 26
7 16
42 21
0 21
42 22
0 13
0 17
10 15
45 27
3 21
47 21
1 28
42 22
0 13
0 IS
17 14
2 3
20 18
4 5
24 23
0 0
24 22
0 1
0 0
19 19
0 0
18 22
5 0
22 22
1 0
23 22
0 0
0 0
EnNormal larged
Normal
1
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Control Contact Inoculated
the pale center and under normal conditions would supply sufficient quantities of erythrocytes. As indicated in Table 6, the marrow observed in some of the infected birds was red throughout. Since in affected birds the hemoglobin content is reduced as well as the number of red cells, the demand for erythrocytes is great. As a result, the area of erythropoiesis was greatly extended and appeared to involve the entire marrow. In approximately 50 percent of the infected birds the kidneys were enlarged and often pale. Enlarged and/or discolored livers among the infected birds were observed frequently. In advanced cases a greenish discoloration of the liver was more common than in less severe cases. In general observations on the internal organs as a means of diagnosis can be very misleading. The stage of the infection has much to do with the pathological changes of the organs. This is illustrated by the splenomegaly which was more common in the early stages, whereas kidney and liver enlargement were more frequent to the advanced stages. Duration of Antibiotic Treatment. Preliminary information concerning the length of time which antibiotics must be
CONTROL OF INFECTIOUS SYNOVITIS
119
TABLE 7.—Occurrence of infectious synovitis as influenced by antibiotic treatment Treatment Group No. 1 2 3 4 5 6
Controls—not previously exposed to infection Aureomycin—continuous Terramycin—continuous Aureomycin—discontinued at 7 weeks Terramycin—discontinued at 7 weeks Aureomycin—started after signs—discontinued at 7 weeks Terramycin—started after signs—discontinued at 7 weeks
fed to experimentally infected birds to prevent the infection from occurring was obtained by continued observations on certain birds used to obtain the data presented above. At this time all birds were seven weeks old and as shown in Table 7, only two birds had signs of infection. Those birds were in group 7 in which terramycin had been provided after signs of synovitis had appeared. Birds in group 1 had received a commercial ration and had not previously been exposed by contact or experimental infection. The previous and subsequent treatment of groups 2 through 7 are indicated in Table 7. For example, group 2 had received 200 grams of aureomycin per ton of feed since they were a week old. This treatment was continued for the additional period of three weeks at which time the birds were 10 weeks old. For these groups, birds were selected so as to obtain equal numbers of those which had been previously inoculated at 23 days of age and those which had not been inoculated. Reoccurrence of signs of infectious synovitis was apparent in groups 5 to 7. Continuous feeding of aureomycin and terramycin was effective in preventing development of the condition as shown in groups 2 and 3. The continuous feeding
No. birds
Age, weeks 7
8
9
10
6 12 12 12 12
0 0 0 0 0
0 0 0 0 1
0 0 0 0 1
0 0 0 0 2
6
0
2
2
2
6
2
3
3
4
of aureomycin for six weeks and discontinued at seven weeks, group 4, prevented the reoccurrence of any signs of the disease. Similar treatment with terramycin, group 5, did not afford the same protection. In this group two birds developed the disease. One was previously inoculated and the other was a contact control. In groups 6 and 7 where signs of synovitis were evident before the antibiotics were given, the removal of either drug resulted in a reoccurrence of the infection. Signs reappeared within a week in two of the three birds of group 6 which had been previously inoculated. In the terramycin-treated group all three previously inoculated birds and one of three contact controls became infected. The incubation period of infectious synovitis in contact control chickens has been approximately 30 days or longer (Olson et al., 1956). It appeared that the contact infection must have occurred prior to the time when the terramycin feeding was discontinued. From the results obtained it seems evident that neither aureomycin nor terramycin completely destroyed the infectious synovitis agent once a build-up of the disease had taken place. Aureomycin
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7
Description
Occurrence of lissions
120
SHELTON, BLETNER, OLSON, ANDERSON AND WEAKLEY, JR.
treatment brings about a regression of signs if treatment is started immediately upon their appearance. Terramycin was less effective. SUMMARY
ACKNOWLEDGEMENT
The authors are indebted to the following concerns for material gratuitously supplied during the course of this investigation: Abbott Laboratories, North Chicago, Illinois for crystalline erythromycin; Chas. Pfizer & Co., Inc., Terre Haute, Indiana for crystalline terramycin hydrochloride; Lederle Laboratories Division, American Cyanamid Company,
REFERENCES Bletner, J. K., D. C. Shelton, N. O. Olson and C. E. Weakley, Jr., 1955. Synovitis control. 3. The efficacy of aureomycin with relation to time of inoculation. Poultry Sci. 34:1181. Cover, M. S., and J. N. Galeta, 1955. Studies on avian arthritis of chickens. Proceedings 27th Annual Pullorum Conference, University of New Hampshire, Durham, New Hampshire. Cover, M. S., J. N. Galeta and E. F. Waller, 1956. The etiology of an arthritic disease of chickens. Am. J. Vet. Res. 17: 12-15. Olson, N. O., J. K. Bletner, D. C. Shelton, D. A. Munro and G. C. Anderson, 1954a. Enlarged joint condition in poultry caused by an infectious agent. Proceedings of West Virginia Feed Dealer's Short Course, April 19,1954. Olson, N. O., J. K. Bletner, D. C. Shelton, D. A. Munro and G. C. Anderson, 1954b. Enlarged joint condition in poultry caused by an infectious -•• agent. Poultry Sci. 33:1075. Olson, N. O., J. K. Bletner, D. C. Shelton, D. A. Munro and G. C. Anderson, 1955. Infectious anemia-synovitis. Science Serves Your Farm, Annual Report published quarterly, Agric. Exp. Sta., West Virginia University. Olson, N. O., D. C. Shelton, J. K. Bletner, D. A. Munro and G. C. Anderson, 1956. Studies of infectious synovitis in chickens. Am. J. Vet. Res. 17:747. Olson, N. O., D. C. Shelton, J. K. Bletner and C. E. Weakley, Jr., 1955. Synovitis control. 2. A comparison of levels of antibiotics. Poultry Sci. 34: 1214. Shelton, D. C , J. K. Bletner, N. O. Olson, G. C. Anderson and C. E. Weakley, Jr., 1955. Synovitis control. 1. Continuous feeding of antibiotics and the influence of stilbestrol and coccidiostats. Poultry Sci. 34: 1221. Snoeyenbos, G. H., and O. M. Olesiuk, 1955. Studies of an agent producing arthritis in turkeys. Proceedings 27th Annual Pullorum Conference, University of New Hampshire, Durham, N. H. Wills, F. K., 1954. Preliminary report on transmission of an agent producing an arthritis in chick-
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The use of crystalline aureomycin, terramycin and erythromycin in the control of infectious synovitis has been investigated under laboratory conditions. Erythromycin was ineffective against the infectious agent used. However, 200 grams of aureomycin or terramycin per ton of feed was effective when fed continuously. Terramycin appeared less effective when treatment was delayed until after the birds showed signs of synovitis. Hemoglobin values were lowered substantially in infected birds. There were marked changes in the differential white cell counts. Infected birds showed a reduced percentage of lymphocytes, but an increased percentage of heterophils, monocytes and immature forms of leucocytes. Observations were made on internal organs. The pathological changes of these organs are influenced by the stage of the infection. The use of diethylstilbestrol, sulfaquinoxaline, nitrophenide, sulfamethazine or combinations of these drugs had no detrimental or therapeutic influence on the course of the disease.
Pearl River, New York for crystalline aureomycin hydrochloride; Merck and Co., Inc., Rahway, New Jersey for Bcomplex vitamins and menadione; and Nopco Chemical Company, Harrison, New Jersey for vitamin A (Nopcay 10) and vitamin D 3 (Super Nopdex 15).
CONTROL OF INFECTIOUS SYNOVITIS ens. Progress Report 1674. Texas Agric. Exp. Sta., College Station, Texas, April 26,1954. Wills, F. K., 1955. Studies of an unidentified agent producing arthritis in chickens. Southwestern Vet. 8: Part 2.
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Wills, F. K., and J. P. Delaplane, 1955. Transmission and therapy on an agent which produces arthritis in chickens. Proc. 92nd meeting, Am. Vet. Med. Assoc. 350-356.
The Efficacy of Chlortetracycline at Several Temperatures in Controlling Spoilage of Poultry Meat* W. G. SHANNON1 AND W. J. STADELMAN (Received for publication July 16, 1956) INTRODUCTION
T
HE volume of poultry meat moving through market channels is increasing each year. A high percentage of this meat is fryer chickens most of which are delivered to the final consumer without freezing. One of the major problems in handling the fresh fryers is the short keeping time. The spoilage of fresh poultry meat held under refrigeration without freezing is predominantly bacterial. This information has been available for years. Lockhead and Landerkin (1935) reported that spoilage of chicken held at refrigerator temperatures above freezing was due primarily to surface bacteria. Further confirmation comes from the work of Ayres, Ogilvy and Stewart (1950). The appearance of slime nodules and off-odor has been commonly used to determine spoilage, but Ziegler, Spencer and Stadelman (1954) report a rapid microscopic method for this determination. This was the method used in this study.
On November 30, 1955, the Federal Food and Drug Administration cleared chlortetracycline for use "in and on uncooked poultry." Chlortetracycline is * Journal Paper No. 1001 of the Purdue University Agricultural Experiment Station. 1 Present Address: Loughry Agricultural College, Cookstown County, Tyrone, North Ireland.
recognized as one of the most effective of the broad spectrum antibiotics and within recent years a vast amount of work has been done in both the fields of human medicine and agriculture on the use of chlortetracycline. Ziegler and Stadelman (1955) report on the effectiveness of chlortetracycline in the retardation of spoilage of poultry meat while Kohler, Miller and Broquist (1955) report chlortetracycline being the most effective of several antibiotics which they used. The purpose of this paper is to report on the effect of chlortetracycline treatment of eviscerated poultry held at different levels of temperature. EXPERIMENTAL PROCEDURE
The study consisted of two treatments at five temperature levels of storage. It was conducted in two equal parts. In the first the three storage temperatures used were 32°, 37°, and 42°F. and in the'second 42°, 47°, and 68°F. There were two replicates at each temperature and six birds in each replicate. The birds used were fryers which were dressed using a batch scalder at 140°F. for 40 seconds. After picking on a mechanical picker they were cooled in ice water, eviscerated and then further cooled in ice. Each bird was divided into halves, one half being used in treatment 1, the other in treatment 2.
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Purdue University, Agricultural Experiment Station, Lafayette, Indiana