Animal & Veterinary
NADA 140-867 BIO-COX® + AUREOMYCIN® + 3-Nitro® - original approval
Approval Date: June 12, 1989
I. GENERAL INFORMATION:
| NADA | 140-867 |
| Sponsor: | American Cyanamid Company P.O. Box 400 Princeton, N.J. 08540 |
| Generic Name: | salinomycin sodium + chlortetracycline + roxarsone |
| Trade Name: | BIO-COX® + AUREOMYCIN® + 3-Nitro® |
| Marketing Status: |
II. Indications for Use:
For the prevention of coccidiosis in broiler chickens cause by Eimeria acervulina, E. tenella, E. brunetti, E. maxima, E. necatrix and E. mivati including some field strains of E. tenella that are more susceptible to roxarsone combined with salinomycin than salinomycin alone as an aid in the reduction of mortality due to E. coli infections susceptible to such treatment.
III. DOSAGE
| A. | DOSAGE FORM |
Finished feeds manufactured from: BIO-COX - supplied as a medicated premix in 50 pound bags. Each pound of premix contains 30 g of salinomycin sodium activity. AUREOMYCIN 50, 70 and 100 Granular Type A Medicated Articles contain chlortetracycline calcium complex equivalent to 50, 70 and 100 grams of chlortetracycline HCl per pound of premix, respectively. 3-Nitro 10, 20 or 50 medicated premix containing 10, 20 or 50% activity, respectively. |
| B. | ROUTE OF ADMINISTRATION | Oral administration via the feed. |
| C. | RECOMMENDED DOSAGES: |
Salinomycin sodium (from BIO-COX) at concentrations ranging from 0.0044% to 0.0066% (40-60 g/ton) in finished feed. Chlortetracycline (from AUREOMYCIN) at a concentration of 0.055% (500 g/ton in finished feed. |
IV. EFFECTIVENESS
The approved original NADA 128-686 for BIO-COX (salinomycin sodium) (48 FR 30616; 7/5/83) contains adequate data to establish that salinomycin sodium at a range of 0.0044% to 0.0066% (40-60 g/ton) in finished feed is safe and effective for the prevention of coccidiosis caused by Eimeria acervulina, E. tenella, E. brunetti, E. maxima, E. necatrix and E. mivati.
The approved NADA for salinomycin plus roxarsone, NADA 132-447, contains adequate data to established that the addition of roxarsone (3-Nitro-4-hydroxyphenylarsonic acid) at a concentration of 0.005% (45.4 g/ton) + salinomycin sodium between the range of 0.0044 to 0.0083% (40-75 g/ton) is safe and effective for the prevention of coccidiosis in broiler chickens caused by Eimeria tenella, E. necatrix, E. acervulina, E. maxima, E. brunetti and E. mivati , including some field strains of E. tenella that are more susceptible to roxarsone combined with salinomycin than the salinomycin alone.
The approved original NADA 48-761, AUREOMYCIN chlortetracycline (32 FR 14155; 10/12/67) contained adequate data to establish that at concentrations of 500 g/ton in finished feed the drug is safe and effective as an aid in the reduction of mortality due to E. coli infections susceptible to such treatment.
Data supplied in this submission demonstrate that the combination of salinomycin sodium at concentrations of 0.0044% to 0.0083% (40-75 g/ton) + chlortetracycline at a concentration of 0.055% (500 g/ton) + roxarsone at a concentration of 0.0050% (45.4 g/ton) is effective for the prevention of coccidiosis in broiler chickens caused by E. necatrix, E. acervulina, E. tenella, E. maxima, E. brunetti and E. mivati , including some field strains of E. tenella that are more susceptible to roxarsone combined with salinomycin than salinomycin alone, and as an aid in the reduction of mortality due to E. coli infections susceptible to such treatment.
A. Noninterference in Battery Studies
Experiment A-86-21 E. coli Challenge Study
Four hundred Hubbard x Hubbard broiler chickens at nine days of age were used in an adequate, well-controlled battery study. The study was conducted in a uniform environment with a 12 hour light-dark cycle to test for noninterference of BIO-COX salinomycin and 3-Nitro roxarsone with the antibacterial efficacy of AUREOMYCIN chlortetracycline.
This study was conducted by using the recommended use level of chlortetracycline (500 g/ton). The experimental treatment groups were as follows: uninfected controls (nonmedicated and medicated), infected nonmedicated control, and infected groups fed chlortetracycline at 500 g/ton (550 ppm) with or without salinomycin at 75 g/ton (82.5 ppm) and/or roxarsone at 45.4 g/ton (50 ppm) in the diet.
There were four replicate pens per treatment and each pen contained five males and five females. At 3 hours post-treatment the nine-day-old birds were challenged by injection in the right thoracic air sac with a culture containing approximately 1.4 x 104 cfu per dose of chlortetracycline sensitive E. coli . Noninfected control birds were infected similarly with sterile phosphate buffered saline. The birds were medicated for 5 days and a 7 day withdrawal period ensued. Feed consumption and mortality were recorded daily for each pen. Birds were weighed at 5 and 12 days post challenge. Results are in Table 1. Survivors were sacrificed 12 and 13 days post challenge. At necropsy, all birds were assigned air sac lesion scores. Air sac swabs were cultured in attempts to recover the challenge E. coli.
The results presented in Table 2 show that chlortetracycline incorporated in the diet at 550 ppm (500 g/ton) prevented the development of E. coli induced air-sac infection in chickens challenged with E. coli . BIO-COX salinomycin and 3-Nitro roxarsone in combination with chlortetracycline did not interfere with the antibacterial efficacy of chlortetracycline in broilers.
The investigators who conducted the above study were:
J.D. Kobland, R.L. Lubas, J.L. Samuel
American Cyanamid Company
Agricultural Research Division
P.O. Box 400
Princeton, New Jersey 08540
(Eds. note: The following table consists of 6 columns.)
Table 1 Experiment A-86-21 Summary of Daily Gain/Chick
Drug Daily Gain/ Daily Gain/
Trt Level E. coli Chick (g) Chick (g)
Group Treatment (ppm) Challenge 0-5 days* 0-12 days
A Non-medicated 0 No 31.6 39.0
B Salinomycin + 82.5
Roxarsone + 50 No 30.0 38.4
Chlortetracycline 550
C Non-medicated 0 Yes 23.7 32.5
D Cholortetracycline 550 Yes 30.6 37.6
E Salinomycin 82.5 Yes 22.2 30.1
F Salinomycin + 82.5
Chlortetracycline 550 Yes 27.4 36.0
G Salinomycin + 82.5 Yes 25.3 34.2
Roxarsone 50
H Roxarsone 50 Yes 26.5 34.8
I Roxarsone + 50 Yes 28.8 37.0
Chlortetracycline 550
J Salinomycin + 82.5 Yes 28.8 37.5
Roxarsone + 50
Chlortetracycline 550
* Treatment period
(Eds. note: The following table consists of 8 columns.)
Table 2 Experiment A-86-21 Summary of Mortality, Average Air Sac Scores, E. coli Recovery Scores and Incidence of Chickens Positive for E. coli
Av. No.Chickens
Drug No. Av. E. coli Positive
Trt. Level E. coli Dead/ Lesion Recovery For E. coli
Group Treatment (ppm) Challenge Total* Score** Score Total***
A Non-Medicated 0 No 0/40 0.05 0.00 0/40
B Salinomycin 82.5
Roxarsone 50 No 0/40 0.00 0.00 0/40
Chlortetracycline 550
C Non-Medicated 0 Yes 9/40 1.80 1.19 17/40
D Chlortetracycline 550 Yes 1/38 0.60 0.07 2/38
E Salinomycin 82.5 Yes 6/39 1.70 0.67 8/39
F Salinomycin + 82.5 Yes 0/40 1.12 0.15 4/40
Chlortetracycline 550
G Salinomycin 82.5 Yes 10/40 1.65 1.02 11/40
Roxarsone 50
H Roxarsone 50 Yes 8/40 1.58 0.86 11/40
I Roxarsone 50 Yes 0/40 1.28 0.00 0/40
Chlortetracycline 550
J Salinomycin 82.5 Yes 0/40 0.98 0.02 1/40
Roxarsone 50
Chlortetracycline 550
* Number of chicks which died due to induced E. coli infection per total chicks (chicks which died at challenge or accidentally were excluded from these data).
** Mean scores were computed from pen averages.
*** Excluding birds which died at challenge.
B. Coccidial Challenge Studies: Two week old Hubbard x Hubbard broiler chickens were used in adequate, well-controlled battery studies. The studies were conducted in a uniform environment with continuous artificial illumination to test for noninterference of AUREOMYCIN chlortetracycline with the anticoccidial effectiveness of BIO-COX salinomycin in combination with 3-Nitro roxarsone. Recent field strain isolates collected from various broiler producing regions of the United States were used. For each study, combinations of E. acervulina/E. mivati, E.necatrix and E. brunetti or E. acervulina, E. maxima and E. tenella were used. Combinations of three challenge species in each study were used to differentiate areas of the intestines infected by the parasite.
These studies were conducted by using the lowest recommended use level of salinomycin (40 g/ton). The experimental treatment groups were as follows:
(Eds. note: The following table consists of 4 columns.)
Treatment -------Medication--------
Group Drug g/ton Infection
I None None None
II None None Yes
III Salinomycin 40 Yes
IV Chlortetracycline 500 Yes
V Salinomycin + 40 Yes
Chlortetracycline 500
VI Roxarsone + 45 Yes
Chlortetracycline 500
VII Salinomycin + 40 Yes
Roxarsone 45
Chlortetracycline 500
VIII Salinomycin + 40 Yes
Roxarsone 45
IX Roxarsone 45 Yes
In each of these studies all pens were preselected, birds were randomized by weight and assigned to cages with ten birds/cage, and four replicates were used per treatment group. Groups were medicated two days before infection was administered. Evaluations were by lesion scores, (analysis was performed on preselected birds from each pen), dropping scores, weight gains and feed conversions. Salinomycin at 40 g/ton in combination with (3-Nitro) roxarsone at 45 g/ton in combination with (AUREOMYCIN) chlortetracycline at 500 g/ton provided better anticoccidial activity than that obtained with either chlortetracycline alone or unmedicated controls.
The investigators who conducted the above studies were as follows:
S.E. Cheng, D.V.M., Ph.D.
Michael D. Sims, B.S.
Kent L. Puffenbarger, B.S., M.S.
Patricia C. Gerber, A.A.S.
A.H. Robins Co.
Research and Development Division
1211 Sherwood Ave.
Richmond, VA 23220
(Eds. note: The following table consists of 11 columns.)
Table 3 Anticoccidial Activity of Salinomycin in Combination with Chlortetracycline and Roxarsone Against a Mixed Eimeria Infection* in Two Week Old Chicks (Study 85-217)
Table 3 Anticoccidial Activity of Salinomycin in Combination with Chlortetracycline and Roxarsone Against a Mixed Eimeria Infection* in Two Week Old Chicks (Study 85-217) Average Coccidiosis Dropping Treatment Medication Induced Scores** ------Weight Gain (Average/Pen)----- Feed Conversion Total Group Infection Mortality D-4 - D-8 Day 5 Day 6 Day 7 Day 14 Day 7 Day 14 Lesion Scores*** I None 0/40 0.0 1823.2 2234.7 2558.7 6359.7 1.59 1.65 0.0 Uninfected II None 5/40 2.7 1053.4 1022.5 1318.3 4745.1 2.21 2.08 5.7 Mixed III Salinomycin 40 g/ton 0/40 1.4 1431.4 1623.1 1963.8 5369.7 1.82 1.88 4.1 Mixed IV Chlortetracycline 500 g/ton 0/40 2.1 1033.8 1481.1 1914.9 5221.4 1.79 1.83 3.2 Mixed V Salinomycin 40 g/ton + 0/40 0.4 1606.8 2019.3 2408.8 5854.4 1.63 1.72 2.1 Chlortetracycline 500 g/ton Mixed VI Roxarsone 45 g/ton + 0/40 1.8 1262.0 1545.7 1932.5 5216.1 1.80 1.85 3.2 Chlortetracycline 500 g/ton Mixed VII Salinomycin 40 g/ton + 0/40 0.2 1550.2 1904.5 2188.8 5499.2 1.72 1.79 2.5 Roxarsone 45 g/ton + Chlortetracycline 500 g/ton Mixed VIII Salinomycin 40 g/ton 0/40 0.5 1539.7 1844.2 2077.8 5277.9 1.82 1.88 2.7 Roxarsone 45 g/ton Mixed IX Roxarsone 45 g/ton 1/40 2.6 1159.2 1314.2 1671.3 5064.3 2.02 1.95 4.0 Mixed
Note: Treatment VII provided superior (P 0.05) lesion scores over bothTreatment IV and Treatment II.
* E. acervulina, Log No. 984B-1, Cullman, AL, 75,000 sporulated oocysts;
E. maxima, Log No. 938A-6, Ellijay, Ga, 20,000 sporulated oocysts;
E. tenella, Log No. 714B-11, Broadway, VA, 35,000 sporulated oocysts.
** Pen dropping scores assigned using a scale of 0 to 4 (Moorehouse and Baron, 1970).
*** Lesion scores of 0 to 4 were determined for three areas of the small intestine plus ceca with maximum possible total lesion score of 16. (Johnson and Reid, 1970).
(Eds. note: The following table consists of 11 columns.)
Table 4 Anticoccidial Activity of Salinomycin in Combination with Chlortetracycline and Roxarsone Against a Mixed Eimeria Infection* in Two Week Old Chicks (Study 85-218)
Average
Coccidiosis Dropping
Treatment Medication Induced Scores** ------Weight Gain (Average/Pen)----- Feed Conversion Total
Group Infection Mortality D-4 - D-8 Day 5 Day 6 Day 7 Day 14 Day 7 Day 14 Lesion Scores***
I None 0/40 0.0 1772.2 2086.0 2436.0 5618.6 1.48 1.56 0.0
Uninfected
II None 3/40 2.05 929.8 672.3 858.6 3183.3 2.46 2.63 4.7
Mixed
III Salinomycin 40 g/ton 0/40 0.0 1845.7 2270.0 2443.7 5266.4 1.57 1.66 0.7
Mixed
IV Chlortetracycline 500 g/ton 0/40 0.9 1246.5 1625.2 1958.7 4320.5 1.62 1.98 3.3
Mixed
V Salinomycin 40 g/ton + 0/40 0.5 1782.5 2202.7 2464.2 5297.9 1.54 1.65 1.2
Chlortetracycline 500 g/ton
Mixed
VI Roxarsone 45 g/ton + 0/40 0.7 1346.0 1642.7 1963.5 4717.9 1.60 1.90 2.7
Chlortetracycline 500 g/ton
Mixed
VII Salinomycin 40 g/ton + 0/40 0.0 1866.5 2292.5 2507.2 5319.6 1.53 1.65 1.1
Roxarsone 45 g/ton +
Chlortetracycline 500 g/ton
Mixed
VIII Salinomycin 40 g/ton 0/40 0.0 1715.5 2143.0 2366.0 4867.1 1.50 1.79 1.3
Roxarsone 45 g/ton
Mixed
IX Roxarsone 45 g/ton 0/40 1.5 1031.5 1138.7 1281.7 3827.5 1.91 2.44 4.7
Mixed
Note: Treatment VII provided superior (P 0.05) lesion scores over both Treatment IV and Treatment II.
* E. acervulina/mivati, Log No. 984B-1, Cullman, AL, 100,000 sporulated oocysts;
E. necatrix, Log No. 325A-73, Green Co., Ga, 20,000 sporulated oocysts;
E. brunetti, Log No. 237A-51, Madison, WI, 40,000 sporulated oocysts.
** Pen dropping scores assigned using a scale of 0 to 4 (Moorehouse and Baron, 1970).
*** Lesion scores of 0 to 4 were determined for three areas of the small intestine plus ceca with maximum possible total lesion score of 16. (Johnson and Reid, 1970).
V. ANIMAL SAFETY
The original approved NADA's for salinomycin (NADA 128-686), roxarsone (NADA 7-891) and for chlortetracycline (NADA 48-761) contained adequate data to establish the safety of each drug for broiler chickens.
The safety of the three drugs when used in combination was demonstrated in the noninterference battery and coccidial challenge studies described in this document. In each of the several studies there was no evidence that drug was the cause of any mortality or toxicity.
Based on the data in the parent NADA's and from the noninterference battery and coccidial challenge studies, the combination is safe to be fed to broiler chickens as indicated on the label.
The data show that the combination of BIO-COX salinomycin at 40-60 g/ton plus AUREOMYCIN chlortetracycline at 500 g/ton plus roxarsone at 45.4 g/ton in the feed of broiler chickens fulfills all the requirements for a combination drug for animals as follows:
A. Each drug component makes a contribution to the claimed effects;
B. The dosages of each drug component are such that the combination is safe and effective; and
C. The label claims are not antagonistic.
VI. HUMAN SAFETY
A. Data to Support Human Safety
The safety of the approved products, salinomycin sodium (BIO-COX) roxarsone (3-Nitro) and chlortetracycline (AUREOMYCIN) has been established by data submitted in their respective original NADA's, NADA 128-686, NADA 7-891, NADA 48-761 and Salsbury Master File 19.
The tolerance for residues of chlortetracycline in edible tissue of chicken is established at 4.0 ppm in kidney and 1.0 ppm in muscle, liver, fat and skin (21 CFR 556.150). The tolerance for residues of arsenic (due to roxarsone) in liver is established at 2 ppm (CFR 556.60). Safe concentrations of total residues of salinomycin in the edible tissues of chickens are 0.6 ppm for muscle, 1.8 ppm for liver and 1.2 ppm for skin/fat. (Safe concentrations refer to the concentrations of total residues considered safe in edible tissues). Total residues are highest in liver tissue. Parent drug salinomycin in skin/fat was established as the marker residue for total residues in liver. An upper limit of 200 ppb (0.2 ppm) for residues of parent drug salinomycin in skin/fat is established for uses of salinomycin with a zero day withdrawal.
B. Residue Depletion/Non-Interference Studies
Residue data supporting the approved individual uses of salinomycin, chlortetracycline and roxarsone and their withdrawal times of 0 day, 1 day and 5 days, respectively, were submitted in their respective original applications (see Part A above).
Drug residue elimination and assay noninterference studies were conducted to establish that each drug in the presence of the other does not exceed its established safe concentration or tolerance, as applicable, at the proposed withdrawal time and that none of the drugs interferes in any of the other's tissue residue assay.
Control chicken muscle, liver, skin and fat were fortified to contain 2.0 ppm salinomycin, 2.0 ppm roxarsone or 0.50 ppm chlortetracycline. The analyses of these samples by the chlortetracycline microbioassay procedure showed quantitative recovery of chlortetracycline either in the presence or absence of salinomycin plus roxarsone.
Control skin/fat tissue homogenates were spiked with combinations of salinomycin (0.7 ppm) roxarsone (2.0 ppm) and chlortetracycline (1.0 ppm). Neither chlortetracycline or roxarsone interfered with the tissue residue assay procedure (HPLC) for salinomycin.
A tissue depletion study was conducted by A.H. Robins Company under protocol 86-0595 at the A.H. Robins Farm in Ashland, VA. Tissues were obtained from broiler chickens fed 1 of the following: 1) unmedicated control diet for 42 days; 2) a diet containing the drug combination of 75 g/ton salinomycin plus 45 g/ton roxarsone plus 200 g/ton chlortetracycline for the first 37 days, then a diet containing the drug combination of 75 g/ton salinomycin plus 45 g/ton roxarsone plus 500 g/ton chlortetracycline from Day 37 to 42. The medicated group was switched on the morning of Day 42 to unmedicated control diet, and birds were sacrificed following drug withdrawal periods of 0 to 7 days. Tissues were collected, and tissue levels of residue drug determined for the respective drug as follows: salinomycin on Day 0 post withdrawal chlortetracycline at Days 0, 3, 5 and 7 post withdrawal and roxarsone at Days 0, 3 and 7 post withdrawal. Tissues from chickens receiving unmedicated feed during the entire study served as controls with tissues being collected on Day 42 of the study.
The target tissue for residue depletion studies for salinomycin is skin/fat. Salinomycin assayed at < .1 ppm in skin/fat, which is below the upper limit of 0.2 ppm for the marker residue.
At withdrawal days of 0, 3, 5 and 7 all samples assayed for chlortetracycline residues were well below 1 ppm, the established tolerance. Kidney tissue was not assayed, as chicken kidney is no longer considered to be edible tissue.
The depletion of arsenic in the tissues was well below 2.0 ppm, the established tolerance, at both 3 and 7 days withdrawal time.
The results of the residue depletion study (Table 5) support a five day withdrawal for this salinomycin chlortetracycline roxarsone combination.
(Eds. note: The following table consists of 6 columns.)
Table 5 Residue Depletion Assay Results (ppm)
Withdrawal ------------------Tissue-------------------
Day Skin w/
Parameter Assay Fat Muscle Liver Fat Pad
Salinomycin
Established
Safe Concentration
(as Determined by marker 0.2
residue
Salinomycin Assay 0 0.1
(highest residue
found in any broiler)
Chlortetracycline
Established Tolerance 1.0 1.0 1.0 1.0
Chlortetracycline
Assay 0 0.415 0.292 0.520 0.048
(highest residue
found in any broiler 3 0.039 0.029 0.047 0.025
5 0.054 0.070 0.053 0.025
7 0.033 0.028 0.033 0.025
Roxarsone
Established Tolerance 2.0
Total Arsenic Assay 0 4.60
(highest residue 3 0.80
found in any broiler) 7 0.38
Sensitivity of the Method: Salinomycin - 0.10 ppm;
Chlortetracycline - 0.025 ppm;
Arsenic - 0.10 ppm
A regulatory analytical method for salinomycin is not required. Practical analytical methods for the determination of tissue residues of chlortetracycline and roxarsone are available in the Food Additives Analytical Manual on display in FDA's Freedom of Information Room (Room 12A-30, 5600 Fishers Lane, Rockville, MD 20857).
VII. AGENCY CONCLUSIONS
The data submitted in support of this NADA satisfy the requirements of Section 512 of the Federal Food, Drug and Cosmetic Act and demonstrate that the drug combination of salinomycin sodium 40-60 g/ton (0.0044-0.0066%) + chlortetracycline 500 g/ton (0.055%) + roxarsone 45 g/ton (.0050%) in finished feed for broilers is safe and effective for the indications stated on the product labeling.
The tolerances for residues of chlortetracycline in edible tissue of chicken are established at 4.0 ppm in kidney and 1.0 ppm in uncooked muscle, liver, fat and skin (CFR 556.150). Safe concentrations of total residues of salinomycin in the edible tissues of chickens are 0.6 ppm for muscle, 1.8 ppm for liver and 1.2 ppm for skin/fat. The tolerance for residues of arsenic (due to roxarsone) is 0.5 ppm in uncooked chicken muscle tissue and 2 ppm in uncooked edible by-product. The data submitted in support of the NADA indicates that the residues of chlortetracycline and arsenic (due to roxarsone) were below the established tolerance in edible tissues and the residues of salinomycin were below the safe concentration in the edible tissues.
The five day withdrawal time before slaughter is satisfactory.
Adequate directions for use of the proposed combination in broiler chickens have been written for the requested claims in this NADA. Products containing salinomycin and chlortetracycline alone, as well as roxarsone are currently available over the counter.
The agency has concluded that approval of this new animal drug combination will not significantly increase human exposure to residue in edible tissues.
VIII. LABELING (Attached)
- Blue Bird Type C Medicated Feed Tag Label
Copies of this label may be obtained by writing to the:
Freedom of Information Office
Center for Veterinary Medicine, FDA
7500 Standish Place
Rockville, MD 20855