- Email: [email protected]
An in Vitro Comparison of Some Antibacterial Agents on a Strain of Avian Pleuropneumonia-like Organisms*
COMPARISON OF ANTIBACTERIAL AGENTS ON
ORGANISMS
1395
REFERENCES Briggs, G. M., 1952. A review of recent developments in poultry nutrition: Vitamin Bi2, antibiotics, and new growth factors. Trans. Amer. Ass. Cereal Chem. 10:31-50. Combs, G. F., 1951. Unidentified factors required for chick growth. Ninth World's Poultry Congress, 2:35-39. Lillie, R. J., J. R. Sizemore and H. R. Bird, 1953. Unidentified factors in poultry nutrition. I. Development of chick assay. Poultry Sci. 32: 855862. Menge, H., G. F. Combs, P. T. Hsu and M. S. Shorb, 1952. Unidentified growth factors required by chicks and poults. I. Studies with chicks using purified diet. Poultry Sci. 31: 237— 247. Morimoto, H., S. Ariyoshi and H. Hoshii, 1954. Effect of arginine, glycine, methionine, and vitamin Bi2 on chick growth. Tenth World's Poultry Congress, 2:142-144.
An in Vitro Comparison of Some Antibacterial Agents on a Strain of Avian Pleuropneumonia-like Organisms* C . H . DOMERMUTH AND E . P . JOHNSON Section of Animal Pathology, Virginia Agricultural Experiment Station, Blacksburg, Virginia (Received for publication May 2, 1955)
N OVO determinations of the effect of / various antibacterial drugs against the agent of infectious sinusitis (IS) of turkeys have indicated that the agent is sensitive to streptomycin (Hitchner, 1949); streptomycin and aureomycin (Groupe and Winn, 1949); terramycin, streptomycin, aureomycin, nemomycin and Chloromycetin (Gross and Johnson, 1953) ;and magnamycin, terramycin, aureomycin and streptomycin (Wong and James, 1953). Similar in ovo determinations using the agent of chronic respiratory disease (CRD) of chickens have indicated that this agent is sensitive to streptomycin * Supported in part by funds provided by the Agricultural Research Service, U.S.D.A. in the Cooperative Project on Chronic Respiratory Disease.
(Delaplane, 1949); aureomycin, terramycin and Chloromycetin (Van Roekel et al., 1952); terramycin, streptomycin, aureomycin, neomycin and Chloromycetin (Gross and Johnson, 1953); and magnamycin, terramycin, aureomycin and streptomycin (Wong and James, 1953). To our knowledge in vitro drug sensitivity tests on pleuropneumonia-like organisms (PPLO) isolated from avian sources have not been conducted. We feel that valuable etiological information, and information concerning the possible treatment of IS of turkeys and of CRD of chickens might be obtained from such a study. The purpose of this paper is, therefore, to report the results of in vitro drug sensitivity tests on a PPLO strain isolated from an avian source.
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
parison with fish meal. Autolyzed cuttle fish, autolyzed sardines, and fish solubles, when fed to chicks, exhibited growth responses that indicated the presence of unidentified growth factors. However, none of these materials was as potent a source of these growth factors as fish meal. Fungi feed gave only a slight growth response. Six koji feeds were tested, but only two of them showed any effect on chick growth. Brewers' yeast, and koji feeds (a, and c) were nearly as effective as fish meal in promoting chick growth. Apparently the potency of unidentified chick growth factors in microbiological products is somewhat dependent upon the type of cultures used.
PPLO
1396
C. H . DOMEEMUTH AND E . P . JOHNSON MATERIALS AND METHODS
DRUG
IO"8
mycin, nf-180 (furazolidone), Chloromycetin, chlortetracycline (aureomycin), neomycin, sulfadiazine, sulfamerazine, sulfamethazine and sulfaquinoxaline were diluted in 10 fold increments ranging from 10 mg. through 0.000,000,01 mg. per ml. Penicillin was diluted in 10 fold increments ranging from 100,000 units through 0.001 unit per ml. All dilutions were made in the medium described above and the final amount of medium in each tube was 10 ml. After the drug dilutions were made, all tubes of media were inoculated with one loopfull of a six-day stock culture of the PPLO using a 4 mm. diameter loop made of 26 gage wire. All tubes were then incubated for six days at 37°C, inspected for presence or absence of turbidity, re-
LOG. MG. DRUG / M L . MEDIUM IO" 4 IO" 3 IO"8 IO -1 10°
10'
MAGNAMYCIN* OXYTETRACYCLINE* STREPTOMYCIN* FURAZOLIDONE* CHLORTET RACYCLINE CHLOROMYCETIN NEOMYCIN SULFADIAZINE
|.**
SULFAMERAZINE
I**
SULFAMETHAZINE
1**
SULFAQUINOXALINE
|**
PENICILLIN FIG. 1. Drug levels required to inhibit the growth of PPLO in vitro. * These drugs were tested and found to kill PPLO. ** Refractory at these concentrations.
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
The PPLO used in this study was the 155th medium passage of the Massachusetts CRD strain #A5967 (received through the courtesy of Dr. E. Jungherr). This culture was propagated in Difco PPLO Enrichment Broth without crystal violet. Solution of this medium was effected by heating it until it began to boil. It was then filtered through Whatman #1 filter paper and adjusted to pH 7.8 before autoclaving. After autoclaving and cooling, 1,000 units of penicillin were added per ml. of medium. Thallium acetate was then added in a concentration of 1 part in 2,000 and finally one percent Difco PPLO Serum Fraction was added. Eleven drugs; namely, magnamycin, oxy tetracycline (terramycin), strepto-
COMPARISON
or
ANTIBACTERIAL AGENTS ON
In order to find out if the four most active drugs killed or merely inhibited the growth of the PPLO, we first determined the quantity of inoculum which would be large enough to initiate the growth of and small enough to prevent carrying over sufficient drug to prevent the growth of the PPLO. We found that an initial inoculum of 0.1 ml. into 10 ml. of medium and a subsequent subculture of 0.5 ml. of the original into 10 ml. of medium was large enough to eliminate drug interference in the second tube. We then determined if the PPLO were killed or inhibited by transferring 0.5 ml. from each of the last tubes of drug dilutions showing no growth to tubes containing 10 ml. each of stock culture medium. The tubes containing the subculture were
ORGANISMS
1397
incubated for 12 days. At the end of this time the tubes showing no growth were inoculated with 0.1 ml. of a six-day-old stock culture of the PPLO, and observed visually and microscopically for presence or absence of growth. Identical determinations were made from duplicate preparations of all drugs used. RESULTS
The stained smears prepared from the tubes containing the least amount of drug and showing no turbidity revealed neither PPLO nor contaminating organisms. The stained smears prepared from the tubes containing the greatest amount of drug and showing turbidity revealed many PPLO organisms and no contaminating organisms. The duplicate sets of dilutions used in the experiment gave identical results. These results were as follows: The amount of drug per ml. which completely inhibited the growth of the PPLO culture was, in the case of magnamycin 0.000,01 mg., terramycin 0.01 mg., streptomycin 0.01 mg., furazolidone 0.01 mg., aureomycin 0.1 mg., Chloromycetin 0.1 mg., neomycin 1 mg., and, penicillin 100,000 units. Ten mg. amounts of the following drugs per ml. exerted no effect on the growth of the culture: sulfadiazine, sulfamerazine, sulfamethazine and sulfaquinoxaline. The PPLO grown in near lethal concentrations of magnamycin, terramycin, streptomycin, and furazolidone were resistant to the same amount of drug before and after the period of exposure. The PPLO-seeded-tubes which contained 0.000,01 mg. of magnamycin, 0.01 of terramycin, 0.01 mg. of streptomycin, and 0.01 mg. of furazolidone revealed no macroscopic growth of the PPLO. The subcultures taken from these tubes produced no growth, thus indicating that the PPLO
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
incubated for six additional days and reinspected for presence or absence of turbidity. At the end of the second period of incubation duplicate smears were prepared from the tubes containing the greastest amount of drug and showing turbidity, stained with Giemsa and Gram stains, and examined. At this time it was apparent that magnamycin, terramycin, streptomycin, and furazolidone were the most active of the drugs which were tested. In order to determine if the PPLO mutate to a higher resistance when exposed to the action of these drugs, dilutions of magnamycin, terramycin, streptomycin, and furazolidone were inoculated with material obtained from the first tubes which showed turbidity in the dilution series previously described. In the case of the magnamycin dilution, e.g., the first tube to show turbidity contained 0.000,001 mg. of the drug. One-tenth ml. of this material was used as inoculum for each of the tubes included in the new series of magnamycin dilutions.
PPLO
1398
C. H. DOMERMUTH AND E. P . JOHNSON
had been killed by the action of the drugs. After twelve days incubation the media into which the subculture had been inoculated was reinoculated with 0.1 ml. of stock PPLO culture. Visible growth occurred in these tubes in three days.
cal factor in the CRD-IS complex (Nelson, 1936; Markham and Wong, 1952), further work is indicated to determine which of these drugs is most active in vivo. Work of this type is currently in progress at this laboratory.
DISCUSSION
SUMMARY
Twelve drugs were tested in vitro for the ability to inhibit the growth of a strain of PPLO (the Massachusetts CRD strain, A 5967) isolated from an avian source. The drugs were found to be active in the following order of diminishing activity: (a) magnamycin, (b) terramycin, streptomycin, furazolidone, (c) aureomycin, Chloromycetin, (d) neomycin, and (e) penicillin. Magnamycin, terramycin, streptomycin and furazolidone were tested and found to kill PPLO. This strain of PPLO did not readily develop resistance to any of these four drugs. Drugs found to be inactive against the agent studied were sulfadiazine, sulfamerazine, sulfamethazine and sulfaauinoxaline. ACKNOWLEDGEMENTS
The authors wish to acknowledge the technical assistance of Mrs. A. P. Tichelaar, M.T./ASCP. The nf-180 used in this experiment was received through the courtesy of Dr. Paul Harwood, Dr. Hess and Clark, Inc., Ashland, Ohio; magnamycin and terramycin, Chas. Pfizer and Co., Inc., New York, N.Y.; streptomycin, penicillin and sulfaquinoxaline, Merck and Co., Inc., Railway, N.J.; neomycin, Upjohn Co., Kalamazoo, Mich.; aureomycin, sulfadiazine and sulfamethazine, Lederle Laboratories, Pearl River, New York; Chloromycetin, Parke, Davis and Co., Detroit, Mich.; and sulfamerazine, Sharp and Dohme, Philadelphia, Pa.
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
Gross and Johnson (1953) found the in oi3o propagated CRD and IS agent to be influenced by drugs in the following order of diminishing sensitivity; terramycin, streptomycin, aureomycin, neomycin, and Chloromycetin. Wong and James (1953) found the in ovo propagated agent of CRD and IS to be influenced by drugs in the following order of diminishing sensitivity; magnamycin, aureomycin, terramycin, streptomycin and Chloromycetin. We found the in vitro propagated strain of PPLO tested in this study to be influenced by drugs in the following order of diminishing sensitivity; (a) magnamycin, (b) terramycin, streptomycin, furazolidone, (c) aureomycin, Chloromycetin, and (d) neomycin. The in ovo sensitivity tests cited are largely in agreement except in respect to aureomycin. The in vitro tests reported herein follow a similar trend. In addition, our data indicate that this strain of PPLO was killed by magnamycin, terramycin, streptomycin and furazolidone, and that it did not readily develop resistance to these drugs. In ovo propagated CRD and IS strains may contain infectious material other than PPLO. Since this is possible, our in vitro studies indicate that field cases of CRD and IS which cannot be successfully treated by the use of antibiotics may be caused by mixed infections. In ovo and in vitro studies clearly indicate that certain drugs have a depressing effect on PPLO. Since it is currently believed that the PPLO is an active etiologi-
ADMINISTRATION OF VACCINES IN DRINKING WATER REFERENCES Delaplane, J. P., 1949. Cultivation of the chronic respiratory disease virus in chick embryos. Proc. 53rd. Annual Meeting of the United States Livestock Sanitary Association: 193-201. Gross, W. B., and E. P. Johnson, 1953. Effect of drugs on the agents causing infectious sinusitis of turkeys and chronic respiratory disease (ail sac infection of chickens) Poultry Sci. 32: 260263.
Hitchner, S. B., 1949. Streptomycin as a treatment
for infectious sinusitis of turkeys. Poultry Sci. 28: 627-629. Markham, F. S., and S. C. Wong, 1952. Pleuropneumonia-like-organisms in the etiology of turkey sinusitis and chronic respiratory disease of chickens. Poultry Sci. 31:902-904. Nelson, J. B., 1936. Coccobacillary bodies in birds infected with a coryza of slow onset. J. Exp. Med. 63:515-522. Van Roekel, H., O. M. Olesiuk and H. A. Peck, 1952. Chronic respiratory disease of chickens. Am. J. Vet. Res. 13: 252-259. Wong, S. C , and C. G. James, 1953. The susceptibility of the agents of chronic respiratory disease of chickens and infectious sinusitis of turkeys to various antibiotics. Poultry Sci. 32: 589-593.
Administration of Newcastle Disease and Infectious Bronchitis Vaccines through the Drinking Water* R. E. LUGINBUHL, E. L. JUNGHERR AND T. W. CHOMIAK Department of Animal Diseases, Storrs Agricultural Experiment Station, University of Connecticut, Storrs, Conn. (Received for publication May 6, 1955)
INTRODUCTION
I
MMUNIZATION of poultry against Newcastle disease (ND) and infectious bronchitis (IB) has become a common practice. There has been a definite trend toward mass immunization using techniques such as spraying and dusting (Crawley and Fahey, 1954; Hitchner and Reising, 1952; Johnson and Gross, 1951; Luginbuhl, 1952; Luginbuhl and Jungherr, 1952; Markham et al., 1954; Van Roekel et ah, 1951). In vaccinating chickens by the intranasal route it was noted that considerable vaccine was swallowed. This observation stimulated a study of mass immunization through the drinking water. The ability of ND virus and IB virus to infect through the digestive tract was * Supported in part by funds provided by Public Laws 733 (9b3) and a grant from Dr. Salsbury's Laboratories, Charles City, Iowa.
determined by placing gelatin capsules containing virus directly into the crop of chickens. The viability of these viruses at room temperature, in a practical dilution for water vaccination, was investigated as well as the effect of the viruses on chickens when administered in the drinking water. EXPERIMENTAL Experiment 1. This experiment was designed to determine whether birds could be infected with IB virus through the digestive tract. Levine (1954) indicated that it was possible to infect chickens with ND through the digestive tract and Texas Experiment Station workers (Delaplane, 1954) reported success in stimulating immunity to ND by adding the virus to the drinking water. Eight chickens, eight weeks old, having no history of respiratory disease were used. Five birds were given 0.2 ml. of IB
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
Groupe, V., and J. D. Winn, 1949. The characteristics of an agent morphologically resembling the chamydozoaceae and causing sinusitis in turkeys. J. Bact. 57: 515-528.
1399
ORGANISMS
1395
REFERENCES Briggs, G. M., 1952. A review of recent developments in poultry nutrition: Vitamin Bi2, antibiotics, and new growth factors. Trans. Amer. Ass. Cereal Chem. 10:31-50. Combs, G. F., 1951. Unidentified factors required for chick growth. Ninth World's Poultry Congress, 2:35-39. Lillie, R. J., J. R. Sizemore and H. R. Bird, 1953. Unidentified factors in poultry nutrition. I. Development of chick assay. Poultry Sci. 32: 855862. Menge, H., G. F. Combs, P. T. Hsu and M. S. Shorb, 1952. Unidentified growth factors required by chicks and poults. I. Studies with chicks using purified diet. Poultry Sci. 31: 237— 247. Morimoto, H., S. Ariyoshi and H. Hoshii, 1954. Effect of arginine, glycine, methionine, and vitamin Bi2 on chick growth. Tenth World's Poultry Congress, 2:142-144.
An in Vitro Comparison of Some Antibacterial Agents on a Strain of Avian Pleuropneumonia-like Organisms* C . H . DOMERMUTH AND E . P . JOHNSON Section of Animal Pathology, Virginia Agricultural Experiment Station, Blacksburg, Virginia (Received for publication May 2, 1955)
N OVO determinations of the effect of / various antibacterial drugs against the agent of infectious sinusitis (IS) of turkeys have indicated that the agent is sensitive to streptomycin (Hitchner, 1949); streptomycin and aureomycin (Groupe and Winn, 1949); terramycin, streptomycin, aureomycin, nemomycin and Chloromycetin (Gross and Johnson, 1953) ;and magnamycin, terramycin, aureomycin and streptomycin (Wong and James, 1953). Similar in ovo determinations using the agent of chronic respiratory disease (CRD) of chickens have indicated that this agent is sensitive to streptomycin * Supported in part by funds provided by the Agricultural Research Service, U.S.D.A. in the Cooperative Project on Chronic Respiratory Disease.
(Delaplane, 1949); aureomycin, terramycin and Chloromycetin (Van Roekel et al., 1952); terramycin, streptomycin, aureomycin, neomycin and Chloromycetin (Gross and Johnson, 1953); and magnamycin, terramycin, aureomycin and streptomycin (Wong and James, 1953). To our knowledge in vitro drug sensitivity tests on pleuropneumonia-like organisms (PPLO) isolated from avian sources have not been conducted. We feel that valuable etiological information, and information concerning the possible treatment of IS of turkeys and of CRD of chickens might be obtained from such a study. The purpose of this paper is, therefore, to report the results of in vitro drug sensitivity tests on a PPLO strain isolated from an avian source.
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
parison with fish meal. Autolyzed cuttle fish, autolyzed sardines, and fish solubles, when fed to chicks, exhibited growth responses that indicated the presence of unidentified growth factors. However, none of these materials was as potent a source of these growth factors as fish meal. Fungi feed gave only a slight growth response. Six koji feeds were tested, but only two of them showed any effect on chick growth. Brewers' yeast, and koji feeds (a, and c) were nearly as effective as fish meal in promoting chick growth. Apparently the potency of unidentified chick growth factors in microbiological products is somewhat dependent upon the type of cultures used.
PPLO
1396
C. H . DOMEEMUTH AND E . P . JOHNSON MATERIALS AND METHODS
DRUG
IO"8
mycin, nf-180 (furazolidone), Chloromycetin, chlortetracycline (aureomycin), neomycin, sulfadiazine, sulfamerazine, sulfamethazine and sulfaquinoxaline were diluted in 10 fold increments ranging from 10 mg. through 0.000,000,01 mg. per ml. Penicillin was diluted in 10 fold increments ranging from 100,000 units through 0.001 unit per ml. All dilutions were made in the medium described above and the final amount of medium in each tube was 10 ml. After the drug dilutions were made, all tubes of media were inoculated with one loopfull of a six-day stock culture of the PPLO using a 4 mm. diameter loop made of 26 gage wire. All tubes were then incubated for six days at 37°C, inspected for presence or absence of turbidity, re-
LOG. MG. DRUG / M L . MEDIUM IO" 4 IO" 3 IO"8 IO -1 10°
10'
MAGNAMYCIN* OXYTETRACYCLINE* STREPTOMYCIN* FURAZOLIDONE* CHLORTET RACYCLINE CHLOROMYCETIN NEOMYCIN SULFADIAZINE
|.**
SULFAMERAZINE
I**
SULFAMETHAZINE
1**
SULFAQUINOXALINE
|**
PENICILLIN FIG. 1. Drug levels required to inhibit the growth of PPLO in vitro. * These drugs were tested and found to kill PPLO. ** Refractory at these concentrations.
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
The PPLO used in this study was the 155th medium passage of the Massachusetts CRD strain #A5967 (received through the courtesy of Dr. E. Jungherr). This culture was propagated in Difco PPLO Enrichment Broth without crystal violet. Solution of this medium was effected by heating it until it began to boil. It was then filtered through Whatman #1 filter paper and adjusted to pH 7.8 before autoclaving. After autoclaving and cooling, 1,000 units of penicillin were added per ml. of medium. Thallium acetate was then added in a concentration of 1 part in 2,000 and finally one percent Difco PPLO Serum Fraction was added. Eleven drugs; namely, magnamycin, oxy tetracycline (terramycin), strepto-
COMPARISON
or
ANTIBACTERIAL AGENTS ON
In order to find out if the four most active drugs killed or merely inhibited the growth of the PPLO, we first determined the quantity of inoculum which would be large enough to initiate the growth of and small enough to prevent carrying over sufficient drug to prevent the growth of the PPLO. We found that an initial inoculum of 0.1 ml. into 10 ml. of medium and a subsequent subculture of 0.5 ml. of the original into 10 ml. of medium was large enough to eliminate drug interference in the second tube. We then determined if the PPLO were killed or inhibited by transferring 0.5 ml. from each of the last tubes of drug dilutions showing no growth to tubes containing 10 ml. each of stock culture medium. The tubes containing the subculture were
ORGANISMS
1397
incubated for 12 days. At the end of this time the tubes showing no growth were inoculated with 0.1 ml. of a six-day-old stock culture of the PPLO, and observed visually and microscopically for presence or absence of growth. Identical determinations were made from duplicate preparations of all drugs used. RESULTS
The stained smears prepared from the tubes containing the least amount of drug and showing no turbidity revealed neither PPLO nor contaminating organisms. The stained smears prepared from the tubes containing the greatest amount of drug and showing turbidity revealed many PPLO organisms and no contaminating organisms. The duplicate sets of dilutions used in the experiment gave identical results. These results were as follows: The amount of drug per ml. which completely inhibited the growth of the PPLO culture was, in the case of magnamycin 0.000,01 mg., terramycin 0.01 mg., streptomycin 0.01 mg., furazolidone 0.01 mg., aureomycin 0.1 mg., Chloromycetin 0.1 mg., neomycin 1 mg., and, penicillin 100,000 units. Ten mg. amounts of the following drugs per ml. exerted no effect on the growth of the culture: sulfadiazine, sulfamerazine, sulfamethazine and sulfaquinoxaline. The PPLO grown in near lethal concentrations of magnamycin, terramycin, streptomycin, and furazolidone were resistant to the same amount of drug before and after the period of exposure. The PPLO-seeded-tubes which contained 0.000,01 mg. of magnamycin, 0.01 of terramycin, 0.01 mg. of streptomycin, and 0.01 mg. of furazolidone revealed no macroscopic growth of the PPLO. The subcultures taken from these tubes produced no growth, thus indicating that the PPLO
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
incubated for six additional days and reinspected for presence or absence of turbidity. At the end of the second period of incubation duplicate smears were prepared from the tubes containing the greastest amount of drug and showing turbidity, stained with Giemsa and Gram stains, and examined. At this time it was apparent that magnamycin, terramycin, streptomycin, and furazolidone were the most active of the drugs which were tested. In order to determine if the PPLO mutate to a higher resistance when exposed to the action of these drugs, dilutions of magnamycin, terramycin, streptomycin, and furazolidone were inoculated with material obtained from the first tubes which showed turbidity in the dilution series previously described. In the case of the magnamycin dilution, e.g., the first tube to show turbidity contained 0.000,001 mg. of the drug. One-tenth ml. of this material was used as inoculum for each of the tubes included in the new series of magnamycin dilutions.
PPLO
1398
C. H. DOMERMUTH AND E. P . JOHNSON
had been killed by the action of the drugs. After twelve days incubation the media into which the subculture had been inoculated was reinoculated with 0.1 ml. of stock PPLO culture. Visible growth occurred in these tubes in three days.
cal factor in the CRD-IS complex (Nelson, 1936; Markham and Wong, 1952), further work is indicated to determine which of these drugs is most active in vivo. Work of this type is currently in progress at this laboratory.
DISCUSSION
SUMMARY
Twelve drugs were tested in vitro for the ability to inhibit the growth of a strain of PPLO (the Massachusetts CRD strain, A 5967) isolated from an avian source. The drugs were found to be active in the following order of diminishing activity: (a) magnamycin, (b) terramycin, streptomycin, furazolidone, (c) aureomycin, Chloromycetin, (d) neomycin, and (e) penicillin. Magnamycin, terramycin, streptomycin and furazolidone were tested and found to kill PPLO. This strain of PPLO did not readily develop resistance to any of these four drugs. Drugs found to be inactive against the agent studied were sulfadiazine, sulfamerazine, sulfamethazine and sulfaauinoxaline. ACKNOWLEDGEMENTS
The authors wish to acknowledge the technical assistance of Mrs. A. P. Tichelaar, M.T./ASCP. The nf-180 used in this experiment was received through the courtesy of Dr. Paul Harwood, Dr. Hess and Clark, Inc., Ashland, Ohio; magnamycin and terramycin, Chas. Pfizer and Co., Inc., New York, N.Y.; streptomycin, penicillin and sulfaquinoxaline, Merck and Co., Inc., Railway, N.J.; neomycin, Upjohn Co., Kalamazoo, Mich.; aureomycin, sulfadiazine and sulfamethazine, Lederle Laboratories, Pearl River, New York; Chloromycetin, Parke, Davis and Co., Detroit, Mich.; and sulfamerazine, Sharp and Dohme, Philadelphia, Pa.
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
Gross and Johnson (1953) found the in oi3o propagated CRD and IS agent to be influenced by drugs in the following order of diminishing sensitivity; terramycin, streptomycin, aureomycin, neomycin, and Chloromycetin. Wong and James (1953) found the in ovo propagated agent of CRD and IS to be influenced by drugs in the following order of diminishing sensitivity; magnamycin, aureomycin, terramycin, streptomycin and Chloromycetin. We found the in vitro propagated strain of PPLO tested in this study to be influenced by drugs in the following order of diminishing sensitivity; (a) magnamycin, (b) terramycin, streptomycin, furazolidone, (c) aureomycin, Chloromycetin, and (d) neomycin. The in ovo sensitivity tests cited are largely in agreement except in respect to aureomycin. The in vitro tests reported herein follow a similar trend. In addition, our data indicate that this strain of PPLO was killed by magnamycin, terramycin, streptomycin and furazolidone, and that it did not readily develop resistance to these drugs. In ovo propagated CRD and IS strains may contain infectious material other than PPLO. Since this is possible, our in vitro studies indicate that field cases of CRD and IS which cannot be successfully treated by the use of antibiotics may be caused by mixed infections. In ovo and in vitro studies clearly indicate that certain drugs have a depressing effect on PPLO. Since it is currently believed that the PPLO is an active etiologi-
ADMINISTRATION OF VACCINES IN DRINKING WATER REFERENCES Delaplane, J. P., 1949. Cultivation of the chronic respiratory disease virus in chick embryos. Proc. 53rd. Annual Meeting of the United States Livestock Sanitary Association: 193-201. Gross, W. B., and E. P. Johnson, 1953. Effect of drugs on the agents causing infectious sinusitis of turkeys and chronic respiratory disease (ail sac infection of chickens) Poultry Sci. 32: 260263.
Hitchner, S. B., 1949. Streptomycin as a treatment
for infectious sinusitis of turkeys. Poultry Sci. 28: 627-629. Markham, F. S., and S. C. Wong, 1952. Pleuropneumonia-like-organisms in the etiology of turkey sinusitis and chronic respiratory disease of chickens. Poultry Sci. 31:902-904. Nelson, J. B., 1936. Coccobacillary bodies in birds infected with a coryza of slow onset. J. Exp. Med. 63:515-522. Van Roekel, H., O. M. Olesiuk and H. A. Peck, 1952. Chronic respiratory disease of chickens. Am. J. Vet. Res. 13: 252-259. Wong, S. C , and C. G. James, 1953. The susceptibility of the agents of chronic respiratory disease of chickens and infectious sinusitis of turkeys to various antibiotics. Poultry Sci. 32: 589-593.
Administration of Newcastle Disease and Infectious Bronchitis Vaccines through the Drinking Water* R. E. LUGINBUHL, E. L. JUNGHERR AND T. W. CHOMIAK Department of Animal Diseases, Storrs Agricultural Experiment Station, University of Connecticut, Storrs, Conn. (Received for publication May 6, 1955)
INTRODUCTION
I
MMUNIZATION of poultry against Newcastle disease (ND) and infectious bronchitis (IB) has become a common practice. There has been a definite trend toward mass immunization using techniques such as spraying and dusting (Crawley and Fahey, 1954; Hitchner and Reising, 1952; Johnson and Gross, 1951; Luginbuhl, 1952; Luginbuhl and Jungherr, 1952; Markham et al., 1954; Van Roekel et ah, 1951). In vaccinating chickens by the intranasal route it was noted that considerable vaccine was swallowed. This observation stimulated a study of mass immunization through the drinking water. The ability of ND virus and IB virus to infect through the digestive tract was * Supported in part by funds provided by Public Laws 733 (9b3) and a grant from Dr. Salsbury's Laboratories, Charles City, Iowa.
determined by placing gelatin capsules containing virus directly into the crop of chickens. The viability of these viruses at room temperature, in a practical dilution for water vaccination, was investigated as well as the effect of the viruses on chickens when administered in the drinking water. EXPERIMENTAL Experiment 1. This experiment was designed to determine whether birds could be infected with IB virus through the digestive tract. Levine (1954) indicated that it was possible to infect chickens with ND through the digestive tract and Texas Experiment Station workers (Delaplane, 1954) reported success in stimulating immunity to ND by adding the virus to the drinking water. Eight chickens, eight weeks old, having no history of respiratory disease were used. Five birds were given 0.2 ml. of IB
Downloaded from http://ps.oxfordjournals.org/ at University of Arizona on June 7, 2016
Groupe, V., and J. D. Winn, 1949. The characteristics of an agent morphologically resembling the chamydozoaceae and causing sinusitis in turkeys. J. Bact. 57: 515-528.
1399