Prophylactical and Therapeutical Efficacy of Tiamuline in Mycoplasmosis of Chickens and Turkeys L. STIPKOVITS1, G. LABER2 and E. SCHULTZE2 1
Veterinary Medical Research Institute, Hungarian Academy of Sciences, Budapest, Hengary, and 2 Sandoz Research Institute, Vienna, Austria
(Received for publication November 15, 1976)
INTRODUCTION
MATERIALS AND METHODS
Respiratory diseases in poultry due to mycoplasma species are still of great importance in intensive poultry breeding. Mycoplasma (M.) gallisepticum, and in several cases M. synoviae, have been established as the causative agents of mycoplasmosis in chickens (Yamamoto and Adler, 1958; Edward and Kanarek, I 9 6 0 ; Vardaman and Yoder, 1969; Ghazikhanian et al, 1973). Infectious sinusitis of turkeys is also caused by M. gallisepticum but pathological lesions of the lower respiratory tract (airsacculitis) are linked with M. meleagridis infection (Bigland and Yamamoto, 1964). In addition, these diseases are frequently complicated by E. coli infection. Recently, a new antibiotic (Tiamuline, Sandoz 81.723 hfu) has been synthesized (Egger and Reinshagen, 1975) which is a derivative of the diterpene antibiotic Pleuromutilin (Kavanagh et al., 1951). This compound is extremely active against gram positive organisms such as streptococci and staphylococci and against mycoplasms (Drews et al., 1975). Studies in vivo of Tiamuline in comparison to Tylosin tartrate revealed the superior efficacy of the former in experimentally induced M. gallisepticum infections in chickens and turkey poults (Laber and Schutze, 1975). To prove the efficacy under practical conditions, field trials were run to protect birds from mycoplasmal infections and to treat diseased birds suffering from mycoplasmosis.
Antibiotics. Tiamuline (81.723 hfu) from Sandoz Ltd., Basel, was used as a 12.5% aqueous solution. Tylosin tartrate was used as the commercially available soluble powder from Elanco Products, Ltd., London. Experimental procedure. Therapeutical tests were run in flocks suffering from mycoplasmosis with administration of the drug via drinking water for 3 days. Trial 1. 750 Shaver laying hens, 5 months old, average body weight 1.05 kg., floor maintenance, were treated with Tiamuline in a concentration of 0.0125%. Trial 2. 1935 Tetra SL hybrid hens, 10 weeks old, average body weight 1.51 kg., floor maintenance, were treated with Tiamuline in a concentration of 0.025%. Trial 3. 1710 White Diamond hybrid turkeys, 7 weeks old, average body weight 1.23 kg., floor maintenance, were treated with Tiamuline in a concentration of 0.025%. Prophylactical tests were run in chickens and turkey poults from mycoplasma infected flocks. The drug was administered via drinking water for 3 days. Trial 4. 1930 Tetra SL hybrid chickens, 1 week old, floor maintenance, were supplied with Tiamuline in a concentration of 0.0125%. Trial 5. 1000 White Diamond hybrid turkey poults, 1 week old, floor maintenance, were supplied with Tiamuline in a concentration of
1209
Downloaded from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
ABSTRACT To evaluate efficacy of Tiamuline in mycoplasmosis of chickens and turkeys under controlled field conditions 2 prophylactical and 3 therapeutical trials were conducted. The compound was administered via drinking water for 3 days in comparison to Tylosin tartrate. In both prophylactical and therapeutical tests, Tiamuline showed a superior efficacy, as measured by clinical symptoms, pathological lesions and microbiological findings. Therefore, Tiamuline can be recommended for treatment of mycoplasmosis in chickens and turkeys in a dosage of 0.025% and in a dosage of 0.0125% for prophylaxis in flocks at risk. Poultry Science 56:1209-1215, 1977
Body weight in kg. (average)
Serolog. investigations HI test neg. titer 1:2 1:4 1:8 1:16 1:32 1:64
Bacteriol. investigations M. gallisepticum Bacteria (E. coli, Streptococci, Staphylococci)
Pathological lesions None Mild Modest Severe
Size of sample for examination
Clinical symptoms in the flock
Size of test group
Parameters measured
Control group 0.05% Tylosin tartrate
1.05
2 3 3 2
4
5
5 5
10
3 1 1 1 4 1.09
10
760 760 Tracheal rales, nasal discharge, gasping, coughing in about 10% of the birds
Test group 0.0125% Tiamuline
Trial 1
TABLE 1. —Extent of disease in chickens before treatment
1
194
Tes 0.0
(Therapeutica
from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
MYCOPLASMOSIS AND TIAMULINE
1211
TABLE 2. — Extent of disease in turkey poults before treatment [Therapeutical tests) Trial 3 Test group 0.025% Tiamulin
Parameters measured Size of test group Clinical symptoms in the
1740 1740 Average losses about 13%, nasal discharge, gasping, sinusitis in about 80% of animals 30 30 4 18 5 3
4 17 5 4
10
11
12
14
23 7
22 8
... 30 1.23
30 1.22
0.0125%. Control. In each experiment a group of birds of the same breed, age, weight and size, kept under identical conditions in the same pen were treated with Tylosin tartrate in a concentration of 0.05% via drinking water for 3 days. Diagnostic parameters. Prior to treatment a sample of birds of each test flock was arbitrarily selected and necropsied for microbiological, pathomorphological and serological examination. In addition, the body weight was determined. During treatment and up to 5 —7 weeks after treatment only clinical observations were made. At the end of the experiment a further sample of birds was necropsied and investigated as described above. Bacteriological and serological investigation. Heart, liver and air sacs of necropsied birds were cultured on blood agar for bacteria. For demonstration of mycoplasmas the contents of the air sacs were cultivated on "Viand Foie" medium (Barber and Fabricant, 1962), medium " B " (Ern«l and Stipkovits, 1973a) and medium " C " (Calquest, 1962). Mycoplasma strains were identified biochemically (Ernri and Stipkovits, 1973b) and serologically by growth inhibition
test (Clyde, 1964) using antisera prepared against all avian mycoplasma species and serogroups (Stipkovits and El-Ebeedy, 1976). Mycoplasma antibodies in chicken sera were determined by slide agglutination and haemagglutination inhibition (HI) test using M. gallisepticum and M. synoviae antigen (Meszaros, 1964). In turkey sera the growth inhibition (GI) test with M. meleagridis antisera was also carried out. Evaluation. The efficacy of the drugs in the therapeutical tests was judged by the course of the clinical symptoms and the occurrence of mycoplasms in the air sacs of necropsied birds. In addition, pathological lesions and the weight gain were considered. For statistical evaluation data from the samples of the different tests were analyzed by a four-fold contingency table. RESULTS Before treatment, mycoplasma infections were confirmed in all flocks assigned for therapeutical trials with Triamuline and Tylosin tartrate by serological, bacteriological and pathological studies (Tables 1 and 2). Mycoplasma
Downloaded from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
Size of sample for evaluation Pathological lesions None Mild Modest Severe Bacteriol. investigations M. meleagridis Bacteria (E. coli, Streptococci, Staphylococci) Serol. investigations M. meleagridis GI test positiv negativ M. gallisepticum HI test positiv negativ Body weight in kg. (average)
flock
Control group 0.05% Tylosin tartrate
Body weight in kg. average 2 weeks after treatment
Bacteriol. investig. M. gallisepticum Bacteria (E. colt)
Pathol, lesions (3 weeks after treatment) None Mild Modest Severe
2 weeks
Clinical symptoms after 1 week
1.16
13 2 2 3
Appetite increasing disappearance of discharge from nostrils and tracheal rales, coughing decreasing Further improvement of symptoms
20
750
Size of test group
Size of sample of birds for specific investigations after treatment
0.0125% Tiamuline
Parameters measured
1.14
1.56
4 1
Further improvement
Symptoms slowly decreasing
4 5 8 3
Gradually decrease of all symptoms of respiratory disease
20
1935
0.025% Tiamuline
No change in comparison to pretreatment symptoms
20
750
0.05% Tylosin tartrate
Trial 1
TABLE 3. — Efficacy of Tiamuline water medication in comparison to Tylosin Tartrate in chi
from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
MYCOPLASMOSIS AND TIAMULINE
DISCUSSION Examinations before treatment indicated the presence of mycoplasma infection in chickens and turkeys assigned for therapeutical tests. The distribution of morbidity rates was the same in the groups treated with Tiamuline and Tylosin tartrate.
The results from each test demonstrate high efficacy of Tiamuline in controlling mycoplasma infection in chickens and turkeys. In therapeutical tests in chickens using Tiamuline in a concentration of 0.0125% (trial 1) an efficacy comparable with Tylosin tartrate given in a dosage of 0.05% was observed. Administration of Tiamuline in a concentration of 0.025% (trial 2) proved superior to Tylosin as indicated by a significant decrease of clinical symptoms, pathological lesions and microbiological findings in birds after treatment. In therapeutical tests in turkeys treated with 0.025% Tiamuline in comparison with 0.05% Tylosin similar results could be obtained. Prophylactical treatment of chickens and turkey poults from mycoplasma infected flocks with Tiamuline prevented higher rate losses, development of pathological lesions as well as bacteriological and mycoplasmal infections than in the control group treated with Tylosin. In addition, higher weight gain could be observed. In conclusion, these data obtained under field conditions show good conformity with data obtained in experimentally induced airsac-
TABLE 4. — Effect of Tiamuline water medication in comparison to Tylosin Tartrate in turkeys (Therapeutical tests) Trial 3 Parameters measured
0.025% Tiamuline
0.05% Tylosin tartrate
Size of test group Size of sample of birds for specific investigation after treatment
1710
1710
39
40
Clinical symptoms after 2 weeks 3 weeks
Decreasing of all symptoms of respiratory disease Further decrease
Statistical evaluation
No change of symptoms Low decrease of symptoms
Pathological lesions (3 weeks after treatment) None Mild Modest Severe
8 1 1
9 9 13
Bacteriol. investigations Mycoplasms (M. meleagridis) Bacteria (E. coli)
7 1
14 18
Body weight in kg. average 2 weeks after treatment
2.22
2.39
Tiamuline superior to tylosin P
Veterinary Medical Research Institute, Hungarian Academy of Sciences, Budapest, Hengary, and 2 Sandoz Research Institute, Vienna, Austria
(Received for publication November 15, 1976)
INTRODUCTION
MATERIALS AND METHODS
Respiratory diseases in poultry due to mycoplasma species are still of great importance in intensive poultry breeding. Mycoplasma (M.) gallisepticum, and in several cases M. synoviae, have been established as the causative agents of mycoplasmosis in chickens (Yamamoto and Adler, 1958; Edward and Kanarek, I 9 6 0 ; Vardaman and Yoder, 1969; Ghazikhanian et al, 1973). Infectious sinusitis of turkeys is also caused by M. gallisepticum but pathological lesions of the lower respiratory tract (airsacculitis) are linked with M. meleagridis infection (Bigland and Yamamoto, 1964). In addition, these diseases are frequently complicated by E. coli infection. Recently, a new antibiotic (Tiamuline, Sandoz 81.723 hfu) has been synthesized (Egger and Reinshagen, 1975) which is a derivative of the diterpene antibiotic Pleuromutilin (Kavanagh et al., 1951). This compound is extremely active against gram positive organisms such as streptococci and staphylococci and against mycoplasms (Drews et al., 1975). Studies in vivo of Tiamuline in comparison to Tylosin tartrate revealed the superior efficacy of the former in experimentally induced M. gallisepticum infections in chickens and turkey poults (Laber and Schutze, 1975). To prove the efficacy under practical conditions, field trials were run to protect birds from mycoplasmal infections and to treat diseased birds suffering from mycoplasmosis.
Antibiotics. Tiamuline (81.723 hfu) from Sandoz Ltd., Basel, was used as a 12.5% aqueous solution. Tylosin tartrate was used as the commercially available soluble powder from Elanco Products, Ltd., London. Experimental procedure. Therapeutical tests were run in flocks suffering from mycoplasmosis with administration of the drug via drinking water for 3 days. Trial 1. 750 Shaver laying hens, 5 months old, average body weight 1.05 kg., floor maintenance, were treated with Tiamuline in a concentration of 0.0125%. Trial 2. 1935 Tetra SL hybrid hens, 10 weeks old, average body weight 1.51 kg., floor maintenance, were treated with Tiamuline in a concentration of 0.025%. Trial 3. 1710 White Diamond hybrid turkeys, 7 weeks old, average body weight 1.23 kg., floor maintenance, were treated with Tiamuline in a concentration of 0.025%. Prophylactical tests were run in chickens and turkey poults from mycoplasma infected flocks. The drug was administered via drinking water for 3 days. Trial 4. 1930 Tetra SL hybrid chickens, 1 week old, floor maintenance, were supplied with Tiamuline in a concentration of 0.0125%. Trial 5. 1000 White Diamond hybrid turkey poults, 1 week old, floor maintenance, were supplied with Tiamuline in a concentration of
1209
Downloaded from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
ABSTRACT To evaluate efficacy of Tiamuline in mycoplasmosis of chickens and turkeys under controlled field conditions 2 prophylactical and 3 therapeutical trials were conducted. The compound was administered via drinking water for 3 days in comparison to Tylosin tartrate. In both prophylactical and therapeutical tests, Tiamuline showed a superior efficacy, as measured by clinical symptoms, pathological lesions and microbiological findings. Therefore, Tiamuline can be recommended for treatment of mycoplasmosis in chickens and turkeys in a dosage of 0.025% and in a dosage of 0.0125% for prophylaxis in flocks at risk. Poultry Science 56:1209-1215, 1977
Body weight in kg. (average)
Serolog. investigations HI test neg. titer 1:2 1:4 1:8 1:16 1:32 1:64
Bacteriol. investigations M. gallisepticum Bacteria (E. coli, Streptococci, Staphylococci)
Pathological lesions None Mild Modest Severe
Size of sample for examination
Clinical symptoms in the flock
Size of test group
Parameters measured
Control group 0.05% Tylosin tartrate
1.05
2 3 3 2
4
5
5 5
10
3 1 1 1 4 1.09
10
760 760 Tracheal rales, nasal discharge, gasping, coughing in about 10% of the birds
Test group 0.0125% Tiamuline
Trial 1
TABLE 1. —Extent of disease in chickens before treatment
1
194
Tes 0.0
(Therapeutica
from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
MYCOPLASMOSIS AND TIAMULINE
1211
TABLE 2. — Extent of disease in turkey poults before treatment [Therapeutical tests) Trial 3 Test group 0.025% Tiamulin
Parameters measured Size of test group Clinical symptoms in the
1740 1740 Average losses about 13%, nasal discharge, gasping, sinusitis in about 80% of animals 30 30 4 18 5 3
4 17 5 4
10
11
12
14
23 7
22 8
... 30 1.23
30 1.22
0.0125%. Control. In each experiment a group of birds of the same breed, age, weight and size, kept under identical conditions in the same pen were treated with Tylosin tartrate in a concentration of 0.05% via drinking water for 3 days. Diagnostic parameters. Prior to treatment a sample of birds of each test flock was arbitrarily selected and necropsied for microbiological, pathomorphological and serological examination. In addition, the body weight was determined. During treatment and up to 5 —7 weeks after treatment only clinical observations were made. At the end of the experiment a further sample of birds was necropsied and investigated as described above. Bacteriological and serological investigation. Heart, liver and air sacs of necropsied birds were cultured on blood agar for bacteria. For demonstration of mycoplasmas the contents of the air sacs were cultivated on "Viand Foie" medium (Barber and Fabricant, 1962), medium " B " (Ern«l and Stipkovits, 1973a) and medium " C " (Calquest, 1962). Mycoplasma strains were identified biochemically (Ernri and Stipkovits, 1973b) and serologically by growth inhibition
test (Clyde, 1964) using antisera prepared against all avian mycoplasma species and serogroups (Stipkovits and El-Ebeedy, 1976). Mycoplasma antibodies in chicken sera were determined by slide agglutination and haemagglutination inhibition (HI) test using M. gallisepticum and M. synoviae antigen (Meszaros, 1964). In turkey sera the growth inhibition (GI) test with M. meleagridis antisera was also carried out. Evaluation. The efficacy of the drugs in the therapeutical tests was judged by the course of the clinical symptoms and the occurrence of mycoplasms in the air sacs of necropsied birds. In addition, pathological lesions and the weight gain were considered. For statistical evaluation data from the samples of the different tests were analyzed by a four-fold contingency table. RESULTS Before treatment, mycoplasma infections were confirmed in all flocks assigned for therapeutical trials with Triamuline and Tylosin tartrate by serological, bacteriological and pathological studies (Tables 1 and 2). Mycoplasma
Downloaded from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
Size of sample for evaluation Pathological lesions None Mild Modest Severe Bacteriol. investigations M. meleagridis Bacteria (E. coli, Streptococci, Staphylococci) Serol. investigations M. meleagridis GI test positiv negativ M. gallisepticum HI test positiv negativ Body weight in kg. (average)
flock
Control group 0.05% Tylosin tartrate
Body weight in kg. average 2 weeks after treatment
Bacteriol. investig. M. gallisepticum Bacteria (E. colt)
Pathol, lesions (3 weeks after treatment) None Mild Modest Severe
2 weeks
Clinical symptoms after 1 week
1.16
13 2 2 3
Appetite increasing disappearance of discharge from nostrils and tracheal rales, coughing decreasing Further improvement of symptoms
20
750
Size of test group
Size of sample of birds for specific investigations after treatment
0.0125% Tiamuline
Parameters measured
1.14
1.56
4 1
Further improvement
Symptoms slowly decreasing
4 5 8 3
Gradually decrease of all symptoms of respiratory disease
20
1935
0.025% Tiamuline
No change in comparison to pretreatment symptoms
20
750
0.05% Tylosin tartrate
Trial 1
TABLE 3. — Efficacy of Tiamuline water medication in comparison to Tylosin Tartrate in chi
from http://ps.oxfordjournals.org/ at North Dakota State University on June 3, 2015
MYCOPLASMOSIS AND TIAMULINE
DISCUSSION Examinations before treatment indicated the presence of mycoplasma infection in chickens and turkeys assigned for therapeutical tests. The distribution of morbidity rates was the same in the groups treated with Tiamuline and Tylosin tartrate.
The results from each test demonstrate high efficacy of Tiamuline in controlling mycoplasma infection in chickens and turkeys. In therapeutical tests in chickens using Tiamuline in a concentration of 0.0125% (trial 1) an efficacy comparable with Tylosin tartrate given in a dosage of 0.05% was observed. Administration of Tiamuline in a concentration of 0.025% (trial 2) proved superior to Tylosin as indicated by a significant decrease of clinical symptoms, pathological lesions and microbiological findings in birds after treatment. In therapeutical tests in turkeys treated with 0.025% Tiamuline in comparison with 0.05% Tylosin similar results could be obtained. Prophylactical treatment of chickens and turkey poults from mycoplasma infected flocks with Tiamuline prevented higher rate losses, development of pathological lesions as well as bacteriological and mycoplasmal infections than in the control group treated with Tylosin. In addition, higher weight gain could be observed. In conclusion, these data obtained under field conditions show good conformity with data obtained in experimentally induced airsac-
TABLE 4. — Effect of Tiamuline water medication in comparison to Tylosin Tartrate in turkeys (Therapeutical tests) Trial 3 Parameters measured
0.025% Tiamuline
0.05% Tylosin tartrate
Size of test group Size of sample of birds for specific investigation after treatment
1710
1710
39
40
Clinical symptoms after 2 weeks 3 weeks
Decreasing of all symptoms of respiratory disease Further decrease
Statistical evaluation
No change of symptoms Low decrease of symptoms
Pathological lesions (3 weeks after treatment) None Mild Modest Severe
8 1 1
9 9 13
Bacteriol. investigations Mycoplasms (M. meleagridis) Bacteria (E. coli)
7 1
14 18
Body weight in kg. average 2 weeks after treatment
2.22
2.39
Tiamuline superior to tylosin P
