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Maternal Transfer of Some Anticoccidial Drugs in the Chicken
RESEARCH NOTES Maternal Transfer of Some Anticoccidial Drugs in the Chicken P. L. LONG, KATHLEEN SHERIDAN, and L. R. McDOUGALD Department of Poultry Science, University of Georgia, Athens, Georgia 30602 (Received for publication December 8, 1980) ABSTRACT Anticoccidial drugs were fed to laying hens for a period of one week. Transfer of drug to the eggs laid by these hens was demonstrated by challenge of the embryos with E. tenella. Maternal transfer of lasalocid, robenidine, and halofuginone occurred. Little or no transfer occurred with arprinocid or monensin. (Key words: anticoccidial drugs, maternal transfer, Eimeria infections of embryos, eoccidia, E. tenella, embryo)
INTRODUCTION The use of chicken embryos for the cultivation of eoccidia was first demonstrated by Long (1965) and has been used for studies on the maternal transfer of antibodies (Rose and Long, 1971) and to demonstrate the maternal transfer of the anticoccidial drugs clopidol, methyl benzoquate, and robenidine (Long, 1971). These drugs are not now as widely used and it seemed worthwhile to study the maternal transfer of several more recently introduced drugs. It is important to know whether anticoccidial drugs given to laying hens in error would be maternally transferred. For experimental studies with eoccidia using chicken embryos, it is vital that anticoccidials are not present. In the present study, hens were given the drugs at concentrations recommended for broilers for one week only and embryos produced by these hens challenged with E. tenella. MATERIALS AND METHODS Fertile eggs were obtained from Single Comb White Leghorn hens. They were placed singly in cages and divided into six groups of six hens each. All hens were housed in the same building. Laying records were kept daily for 10 days before beginning the experimental treatment. The hens were artificially inseminated once weekly with semen collected from roosters housed on the same farm and kept for that purpose. The semen was not diluted and each hen received approximately l/16cm 3 (1 minim). Feed for each group was weighed at the beginning and end of each trial. The differences in feed consumption between trials 1 and 2 were thought to be due to the differences in
ambient temperature between trials 1 and 2. Trial 1 was carried out during May and June, and trial 2 was carried out during July and August when temperatures were much higher. Medicated feed was given ad libitum for 7 days only. Regular and nonmedicated feed was provided ad libitum for the duration of the trial. Details of the treatments are given in Tables 1 and 2. Eggs were collected starting the first day medicated feed was given, providing one day's untreated egg production. Eggs were collected once daily, marked according to the group and date, and placed in cool storage before setting in the incubator at weekly intervals. In the first trial, the eggs were collected for a total of 4 weeks: 1 week on medicated feed and 3 weeks postmedication. In the second trial eggs were collected for only 3 weeks. After the eggs had been incubated at 37 C for 9 days, they were brought to the laboratory and placed in an incubator at 41 C. On day 10 they were candled and injected with 2000 IU penicillin or streptomycin into the allantoic cavity. Sporozoites of E. tenella (WI) strain were frozen and kept in liquid nitrogen until being thawed for use. Each week's group of eggs was inoculated with 100,000 sporozoites/ embryo on day 11 via the allantoic cavity. Effective numbers of embryos were those inoculated minus those dying from nonspecific causes. Embryos that died on day 5 with severe hemorrhage and survivors with severe hemorrhage or lesions were considered positive for coccidiosis.
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RESULTS AND DISCUSSION The details of the experimental groups and
Downloaded from http://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
1981 Poultry Science 60:2342-2345
3 0 11 18 11 36
5 40 11 36 NA
10 40 11 82
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Lasalocid (75)
Robenidine (33)
Arprinocid (60)
Halofuginone (3)
Nonmedicated controls
2
Drug medication ended on day 7.
1 NA = no eggs available. % Positive = those dying with severe hemorrhage and those surviving on 5th PI with se those dying from nonspecific causes before day 5.
10 60
9 11
12 92
7 43
Days 4-72
Infections in eggs collected at differe
Effective # % Positive
Days 0-3
Monensin (120)
Drug used and concentration (ppm)
TABLE 1. Eimeria tenella infections in embryos from hens treated with various (Trial l)1
tp://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
6 100 1 100 4 100 4 100
NA
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Lasalocid (125)
Lasalocid (75)
Arprinocid (60)
Halogufinone (3)
Nonmedicated controls 5 80
12 42
10 60
11 45
7 57
Days 2-6
1 6
4
1 5
1 8
D 7
Infections in eggs collected at differen
2
Drug medication ended on day 7.
1 NA = no eggs available. % Positive = those dying with severe hemorrhage and those surviving on 5th PI with sev those dying from nonspecific causes before day 5.
NA
Effective # % Positive
Days 0-1
Monensin (120)
Drugs used and concentration (PPm)
TABLE 2. Eimeria tenella infections in embryos from hens treated with various a (Trial 2)1
tp://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
RESEARCH NOTE
trations of lasalocid, robenidine, and halofuginone were not detected in embryos ex amined 7 to 12 days after the drug was withdrawn from the hens. REFERENCES Callender, M. E., and R. F. Shumard, 1969. The use of different infection systems for evaluating the efficacy of the anticoccidial compound monensin in chickens housed in floor pens. Poultry Sci. 48: 1792-1793. Long, P. L., 1965. Development of Eimeria tenella in avian embryos. Nature 208:509-510. Long, P. L., 1971. Maternal transfer of anticoccidial drugs in the chicken. J. Comp. Pathol. 81:373382. McManus, E. L., J. E. Brown, D. W. Graham, G. Olson, E. F. Rogers, and T. Tamas, 1976. Anticoccidial battery studies in chickens fed MK-302. Poultry Sci. 55:2065. Mitrivoc, M., and E. G. Schildknecht, 1974. Anticoccidial activity of lasalocid (X-537A) in chicks. Poultry Sci. 53:1448-1455. Rose, M. E., and P. L. Long, 1971. Immunity to coccidiosis: Protective effects of transferred serum and cells investigated in chick embryos infected with Eimeria tenella. Parasitology 6 3 : 299-313. Ruff, M. D., W. I. Anderson, and W. M. Reid, 1978. Effect of the anticoccidial arprinocid on production, sporulation, and infectivity of Eimeria oocysts. J. Parasitol. 64:306-311. Yvore, P., N. Foure, J. Aycardi and G. Bennejean, 1974. Efficacite du Stenerol (RU19110) dans la chimioprophylaxie des coccidioses aviares. Rec. Med. Vet. 150:495-503.
Downloaded from http://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
the results of the two trials which were conducted are given in Tables 1 and 2. All of the anticoccidial agents used in this study are known to be effective against E. tenella (Callender and Shumard, 1969;Mitrovic and Schildknecht, 1974;McManus et al, 1976; Ruff et al, 1978; Yvore et al, 1974). The question here was whether or not these drugs would be passed from hen to egg in a form that would protect the developing embryo from coccidial infection. The results given in Tables 1 and 2 show that apart from monensin, there appears to have been some transfer of each drug in an active form. Lasalocid had the greatest effect in protecting the embryo against challenge with E. tenella. Monesin and lasalocid are both ionophorous antibiotics and may have a similar mode of action on Eimeria. However, there must be differences in the binding of active drug in the hen's egg and embryonic tissues perhaps by the greater affinity for lasalocid to bind to lipids. The amount of drug transferred to the eggs is difficult to estimate, but using the feed intake data and assuming an even distribution of drug throughout the tissues, it appears that the amount of drug consumed in milligrams per kilo body weight was about 9.31, 5.2, 2.97, 6.69, and .29 for monensin (120), lasalocid (75), robenidine (33), arprinocid (60), and halofuginone (3), respectively. Active concen-
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INTRODUCTION The use of chicken embryos for the cultivation of eoccidia was first demonstrated by Long (1965) and has been used for studies on the maternal transfer of antibodies (Rose and Long, 1971) and to demonstrate the maternal transfer of the anticoccidial drugs clopidol, methyl benzoquate, and robenidine (Long, 1971). These drugs are not now as widely used and it seemed worthwhile to study the maternal transfer of several more recently introduced drugs. It is important to know whether anticoccidial drugs given to laying hens in error would be maternally transferred. For experimental studies with eoccidia using chicken embryos, it is vital that anticoccidials are not present. In the present study, hens were given the drugs at concentrations recommended for broilers for one week only and embryos produced by these hens challenged with E. tenella. MATERIALS AND METHODS Fertile eggs were obtained from Single Comb White Leghorn hens. They were placed singly in cages and divided into six groups of six hens each. All hens were housed in the same building. Laying records were kept daily for 10 days before beginning the experimental treatment. The hens were artificially inseminated once weekly with semen collected from roosters housed on the same farm and kept for that purpose. The semen was not diluted and each hen received approximately l/16cm 3 (1 minim). Feed for each group was weighed at the beginning and end of each trial. The differences in feed consumption between trials 1 and 2 were thought to be due to the differences in
ambient temperature between trials 1 and 2. Trial 1 was carried out during May and June, and trial 2 was carried out during July and August when temperatures were much higher. Medicated feed was given ad libitum for 7 days only. Regular and nonmedicated feed was provided ad libitum for the duration of the trial. Details of the treatments are given in Tables 1 and 2. Eggs were collected starting the first day medicated feed was given, providing one day's untreated egg production. Eggs were collected once daily, marked according to the group and date, and placed in cool storage before setting in the incubator at weekly intervals. In the first trial, the eggs were collected for a total of 4 weeks: 1 week on medicated feed and 3 weeks postmedication. In the second trial eggs were collected for only 3 weeks. After the eggs had been incubated at 37 C for 9 days, they were brought to the laboratory and placed in an incubator at 41 C. On day 10 they were candled and injected with 2000 IU penicillin or streptomycin into the allantoic cavity. Sporozoites of E. tenella (WI) strain were frozen and kept in liquid nitrogen until being thawed for use. Each week's group of eggs was inoculated with 100,000 sporozoites/ embryo on day 11 via the allantoic cavity. Effective numbers of embryos were those inoculated minus those dying from nonspecific causes. Embryos that died on day 5 with severe hemorrhage and survivors with severe hemorrhage or lesions were considered positive for coccidiosis.
2342
RESULTS AND DISCUSSION The details of the experimental groups and
Downloaded from http://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
1981 Poultry Science 60:2342-2345
3 0 11 18 11 36
5 40 11 36 NA
10 40 11 82
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Lasalocid (75)
Robenidine (33)
Arprinocid (60)
Halofuginone (3)
Nonmedicated controls
2
Drug medication ended on day 7.
1 NA = no eggs available. % Positive = those dying with severe hemorrhage and those surviving on 5th PI with se those dying from nonspecific causes before day 5.
10 60
9 11
12 92
7 43
Days 4-72
Infections in eggs collected at differe
Effective # % Positive
Days 0-3
Monensin (120)
Drug used and concentration (ppm)
TABLE 1. Eimeria tenella infections in embryos from hens treated with various (Trial l)1
tp://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
6 100 1 100 4 100 4 100
NA
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Effective # % Positive
Lasalocid (125)
Lasalocid (75)
Arprinocid (60)
Halogufinone (3)
Nonmedicated controls 5 80
12 42
10 60
11 45
7 57
Days 2-6
1 6
4
1 5
1 8
D 7
Infections in eggs collected at differen
2
Drug medication ended on day 7.
1 NA = no eggs available. % Positive = those dying with severe hemorrhage and those surviving on 5th PI with sev those dying from nonspecific causes before day 5.
NA
Effective # % Positive
Days 0-1
Monensin (120)
Drugs used and concentration (PPm)
TABLE 2. Eimeria tenella infections in embryos from hens treated with various a (Trial 2)1
tp://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
RESEARCH NOTE
trations of lasalocid, robenidine, and halofuginone were not detected in embryos ex amined 7 to 12 days after the drug was withdrawn from the hens. REFERENCES Callender, M. E., and R. F. Shumard, 1969. The use of different infection systems for evaluating the efficacy of the anticoccidial compound monensin in chickens housed in floor pens. Poultry Sci. 48: 1792-1793. Long, P. L., 1965. Development of Eimeria tenella in avian embryos. Nature 208:509-510. Long, P. L., 1971. Maternal transfer of anticoccidial drugs in the chicken. J. Comp. Pathol. 81:373382. McManus, E. L., J. E. Brown, D. W. Graham, G. Olson, E. F. Rogers, and T. Tamas, 1976. Anticoccidial battery studies in chickens fed MK-302. Poultry Sci. 55:2065. Mitrivoc, M., and E. G. Schildknecht, 1974. Anticoccidial activity of lasalocid (X-537A) in chicks. Poultry Sci. 53:1448-1455. Rose, M. E., and P. L. Long, 1971. Immunity to coccidiosis: Protective effects of transferred serum and cells investigated in chick embryos infected with Eimeria tenella. Parasitology 6 3 : 299-313. Ruff, M. D., W. I. Anderson, and W. M. Reid, 1978. Effect of the anticoccidial arprinocid on production, sporulation, and infectivity of Eimeria oocysts. J. Parasitol. 64:306-311. Yvore, P., N. Foure, J. Aycardi and G. Bennejean, 1974. Efficacite du Stenerol (RU19110) dans la chimioprophylaxie des coccidioses aviares. Rec. Med. Vet. 150:495-503.
Downloaded from http://ps.oxfordjournals.org/ at Karolinska Institutet University Library on June 1, 2015
the results of the two trials which were conducted are given in Tables 1 and 2. All of the anticoccidial agents used in this study are known to be effective against E. tenella (Callender and Shumard, 1969;Mitrovic and Schildknecht, 1974;McManus et al, 1976; Ruff et al, 1978; Yvore et al, 1974). The question here was whether or not these drugs would be passed from hen to egg in a form that would protect the developing embryo from coccidial infection. The results given in Tables 1 and 2 show that apart from monensin, there appears to have been some transfer of each drug in an active form. Lasalocid had the greatest effect in protecting the embryo against challenge with E. tenella. Monesin and lasalocid are both ionophorous antibiotics and may have a similar mode of action on Eimeria. However, there must be differences in the binding of active drug in the hen's egg and embryonic tissues perhaps by the greater affinity for lasalocid to bind to lipids. The amount of drug transferred to the eggs is difficult to estimate, but using the feed intake data and assuming an even distribution of drug throughout the tissues, it appears that the amount of drug consumed in milligrams per kilo body weight was about 9.31, 5.2, 2.97, 6.69, and .29 for monensin (120), lasalocid (75), robenidine (33), arprinocid (60), and halofuginone (3), respectively. Active concen-
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