• Decrease font size
  • Return font size to normal
  • Increase font size
U.S. Department of Health and Human Services

Animal & Veterinary

  • Print
  • Share
  • E-mail

NADA 140-581 Bio-Cox®, 3-Nitro®, and Lincomix® (combined use) - original approval

Approval Date: May 31, 1989

I. GENERAL INFORMATION:

NADA140-581
Sponsor:A. H. Robbins Company, Inc.
1405 Cummings Drive
PO Box 26609
Richmond, VA 23261-6609
Generic Name:salinomycin, roxarsone, and lincomycin
Trade Name:Bio-Cox®, 3-Nitro®, and Lincomix® (combined use)
Marketing Status: 

 

II. Indications for Use:

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 to salinomycin alone; and for improved feed efficiency.

 

III. Dosage Form, Route of Administration and Recommended Dosage:

Type C medicated feeds manufactured from:

  • Bio-Cox - supplied as a Type A medicated article in 50 pound bags. Each pound of Type A medicated article contains 30 g of salinomycin sodium activity.
  • Lincomix - 4, 20 and 50 g of lincomycin activity per pound of Type A medicated article.
  • 3-Nitro - 10, 20 and 50 g of roxarsone per pound of Type A medicated article.

Route of Administration: Oral administration via the feed.

Recommended Dosage: Salinomycin at concentrations ranging from 40 to 60 g/ton, (0.0044 to 0.0066%), plus roxarsone at 45.4% g/ton, plus lincomycin at 2 g/ton, in Type C medicated feed for broiler chickens.

 

IV. Effectiveness:

The parent NADA for salinomycin is 128-686 approved July 5, 1983 (48 FR 30616) at a range of 40 to 60 g/ton for the prevention of coccidiosis caused by Eimeria acervulina, E. brunetti, E. maxima, E. mivati, E. necatrix and E. tenella in broiler chickens. NADA 132-447 is the subject of salinomycin (40-60 g/ton) plus roxarsone (45.4 g/ton) and was approved October 11, 1983 (48 FR 46024) for the prevention of coccidiosis caused by Eimeria acervulina, E. brunetti, E. maxima, E. mivati, E. necatrix and E. tenella, including some field strains of E. tenella that are more susceptible to roxarsone combined with salinomycin than to salinomycin alone. Lincomycin is the subject of NADA 34-085, approved May 9, 1979 (44 FR 7132) for use in broiler chickens at a range of 2 to 4 g/ton for increased rate of weight gain and improved feed efficiency. Data submitted to NADA 140-581 demonstrate that salinomycin plus roxarsone are effective for the prevention of coccidiosis (as described above) and lincomycin at 2 g/ton is effective for improved feed efficiency in broiler chickens.

A. Noninterference Battery Studies

Two week old, Hubbard X Hubbard, broiler chickens were used in adequate, well controlled, two week, battery studies with approved protocols and were conducted in a uniform environment with continuous artificial illumination to test for noninterference of lincomycin and roxarsone with the effectiveness of salinomycin. Recent field strain isolates collected from various broiler producing regions of the United States were used. For each study, combinations of E. mivati, E. brunetti, and E. necatrix, or E. acervulina, E. maxima and E. tenella were used. This arrangement facilitated identification of lesions.

Tables 1 and 2 show the treatments used in these battery studies along with the results for body weight, mortality, lesion scores, and dropping scores. The broilers were randomized by weight and assigned to cages with ten broilers per cage. There were 4 replicates of each treatment group.

These battery studies adequately demonstrate that there is noninterference of lincomycin and roxarsone on the anticoccidial efficacy of salinomycin. Therefore, the combination of salinomycin-roxarsone-lincomycin is compatible.

Ed. note: The following table has 12 columns.

Table 1 Anticoccidial Activity of Salinomycin and Salinomycin Plus Roxarsone in Combination with Lincomycin Against a Mixed Eimeria Infection* in 2 Week Old Chicks (Study 81-57)

Average Average Coccidiosis- Dropping Live-Bird Total Treatment Medication Induced Score** Weight Weight Gain Feed Conversion Lesion Group Infection Mortality D4 - D-8 D14 Day 5 Day 6 Day 7 Day 14 Day 7 Day 14 Score***

I None 0/40 0.0 620 1156 1389 1626 3926 1.55 1.57 0.0 Uninfected II None 6/40 1.9 592 1077 1116 1130 3571 2.04 1.85 6.0 Mixed III 40 g/ton 0/40 0.8 609 1172 1343 1446 3843 1.61 1.63 2.9 salinomycin; mixed IV 4 g/ton 13/40 1.9 583 984 968 1014 3487 2.10 1.89 6.0 linomycin; mixed V 40g/ton 6/40 0.9 618 1133 1239 1323 3894 1.91 1.83 3.1 salinomycin + 4 g/ton lincomycin; mixed VI 4 g/ton 10/40 1.7 567 1074 989 942 3350 2.11 1.85 5.8 linomycin + 45 g/ton roxarson +; mixed VII 40 g/ton 0/40 0.2 643 1234 1418 1597 4106 1.65 1.59 2.2 salinomycin + 45 g/ton roxarsone + 4 g/ton lincomycin; mixed

* E. acervulina, Log No. 3378-1, Washington Co., AR, 25,000 sporulated oocysts; E. maxima, Lot No. 336B-2, Washington Co., AR, 35,000 sporulated oocysts; E. tenella, 249A-5, Marshall Co., AL, 15,000 sporulated oocysts. ** Pen dropping scores assigned using a scale of 0 to 4. *** Lesion scores assigned using a score of 0 to 4 for each area of the small intestine and ceca. Ed. note: The following table has 12 columns. Table II Anticoccidial Activiity of Salinomycin and Salinomycin Plus Roxarsone in Combination with Linomycin Against a Mixed Eimeria Enfection* in 2-Week-Old Chicks (Study 81-58)

Average Average Coccidiosis- Dropping Live-Bird Total Treatment Medication Induced Score** Weight Weight Gain Feed Conversion Lesion Group Infection Mortality D4 - D-8 D14 Day 5 Day 6 Day 7 Day 14 Day 7 Day 14 Score***

I None 0/40 0.0 636 1209 1536 1828 4065 1.49 1.59 0.0 Uninfected II None 1/40 2.2 570 788 831 933 3405 2.20 1.90 5.3 Mixed III 40 g/ton 1/40 0.1 654 1134 1432 1608 4275 1.68 1.66 1.9 salinomycin; mixed IV 4 g/ton 5/40 1.9 575 738 716 856 3439 2.23 1.91 5.7 linomycin; mixed V 40g/ton 0/40 0.1 655 1127 1440 1678 4226 1.63 1.65 2.0 salinomycin + 4 g/ton lincomycin; mixed VI 4 g/ton 1/40 2.2 577 871 880 909 3446 2.08 1.90 6.2 linomycin + 45 g/ton roxarson +; mixed VII 40 g/ton 1/40 2.2 655 1127 1440 1678 4226 1.63 1.65 2.0 salinomycin + 45 g/ton roxarsone + 4 g/ton lincomycin; mixed

* E. mivati, Log No. 328A-2, Talbot, MD, 50,000 sporulated oocysts; E. necatrix, Lot No. 237A-10. Eli Lilly, 100,000 sporulated oocysts; E. brunetti, 325A-9, Greensboro, GA, 100,000 sporulated oocysts. ** Pen dropping scores assigned using a scale of 0 to 4. *** Lesion scores assigned using a score of 0 to 4 for each area of the small intestine and ceca.

The investigators who conducted the above studies were as follows:

Shi E. Cheng, DVM, PhD
A.H. Robins Co.
PO Box 26609
Richmond, VA 23261

Michael D. Sims
A.H. Robins Co.
PO Box 26609
Richmond, VA 23261

Patricia C. Gerber, A.A.S.
A.H. Robins Co.
PO Box 26609
Richmond, VA 23261

B. Floor Pen Studies

The titration data from four trials using four treatment groups were evaluated to determine the effectiveness of this 3 drug combination: treatment 1 contained salinomycin (60 g/ton) plus roxarsone (45.4 g/ton); treatment 2 contained salinomycin at 60 g/ton plus roxarsone at 45.4 g/ton plus lincomycin 2 g/ton; treatment 3 contained salinomycin at 60 g/ton plus roxarsone at 45.4 g/ton plus lincomycin at 3 g/ton; and treatment 4 contained salinomycin at 60 g/ton plus roxarsone at 45.4 g/ton plus lincomycin at 4 g/ton.

The variance at each location for feed efficiency was highly significant, (p<0.0001) indicating that a weighted analysis should be used to evaluate the data. The weighted analysis showed a significant location by treatment interaction (p=0.09) which, when used to test the treatment effect, gave a probability level of 0.106 (0.053 one tailed).

Linear plateau models were fit on the least square means generated in the weighted analysis of the titration data. Model 3-1 had a superior fit, showing that there is a significant response between the control and the 2 g/ton treatment with the responses of all non-zero levels of lincomycin on a plateau. Thus the data are adequate to support lincomycin at 2 g/ton for improved feed efficiency with no significant increase in efficiency with increasing levels of lincomycin.

The data are adequate to support lincomycin at 2 g/ton for improved feed efficiency and for salinomycin at a range of 40-60 g/ton with roxarsone at 45.4 g/ton for the prevention of coccidiosis caused by Eimeria acervulina, E. brunetti, E. maxima, E. mivati, E. necatrix and E. tenella, including some field strains of E. tenella that are more susceptible to roxarsone combined with salinomycin than to salinomycin alone, in broiler chickens.

A summary of the feed efficiency data is presented in Table 3.

Table 3 Summary of Floor Pen Trials

Feed to Gain Data

Trial All treatments contain 60 g/ton Location salinomycin and 45.4 g/ton roxarsone

Treatments 1 2 3 4 Lincomycin: 0 g/ton 2 g/ton 3 g/ton 4 g/ton

Auburn, AL 1.882 1.898 1.878 1.885 Ft. Collins, CO 1.986 1.990 1.972 1.962 Athens, GA 2.029 1.954 1.980 1.985 Richmond, VA 1.947 1.880 1.896 1.878

Location Average: 1.961 1.931 1.931 1.928

Investigators for the floor pen trials:

Dr. Shi E. Cheng
A.H. Robins Co.
Dept. of Vet. Med.
Ashland, VA 23005

Dr. David Rowland
Dept. of Poultry Science
Univ. of Auburn
Auburn, AL 30830

Dr. Larry McDougald
Dept. Poultry Science
Univ. of Georgia
Athens, GA 30602

Dr. Carey Quarles
Colorado Quality Research
1401 Duff Drive
Ft. Collins, CO 80524

The data from the noninterference battery and floor pen studies provide evidence to satisfy the requirements for a combination drug for animals used for human food as follows:

A. Each drug component makes a contribution to the claimed effect.

B. The dosage of each drug is such that the combination is safe.

C. The label claims are not antagonistic or misleading.

 

V. Animal Safety:

The target animal safety data for salinomycin are located in NADA 128-686, approved July 5, 1983 (48 FR 30616). The target animal safety data for roxarsone are located in NADA 7-891, approved March 23, 1951. The target animal safety data for lincomycin are located in NADA 34-085, approved May 9, 1970 (35 FR 7300).

The safety of this drug combination is demonstrated in the four floor pen trials and the noninterference battery trials. The treated birds appeared to be in good health throughout the test period and their growth rate and feed conversion was equal to, or better than, the control birds. Birds that were sacrificed to obtain tissue samples for residue determination showed no signs of lesions or toxicity. Mortality in each study was within an acceptable range for each facility and there was no evidence that drug was the cause of any of the deaths that occurred during any of the studies.

 

VI. Human Food Safety:

A. Data to Support Human Safety

The basic food safety data on which the individual drugs were determined to be safe when used according to their labeling are located as follows: For salinomycin, in NADA 128-686, approved July 5, 1983, (48 FR 30616); for roxarsone, in NADA 7-891, approved March 23, 1951; for lincomycin, in NADA 34-085, approved May 9, 1970 (35 FR 7300). Summaries of these data are on file with Dockets Management Branch (HFA-305) FDA, Room 4-62, 5600 Fishers Lane, Rockville, MD 20857.

B. Residue Depletion & Noninterference Studies

The residue data supporting the approved individual uses of these 3 drugs and their withdrawal periods (salinomycin = 0 days; roxarsone = 5 days; lincomycin = 0 days), have been submitted in their respective parent applications (see Section 6 A. above).

The following studies establish that each drug can be assayed for tissue residue in the presence of the other two drugs without interference from the other two drugs.

  1. Control skin/fat was fortified with 2.0 ppm roxarsone, 0.1 ppm lincomycin and 0.1 ppm salinomycin or with 2.0 ppm roxarsone and 0.1 ppm lincomycin. Examination of the HPLC chromatograms showed that neither roxarsone nor lincomycin interferes with the determination of salinomycin.

    To determine whether roxarsone or salinomycin interfered with the lincomycin assay, stock solutions of each were prepared in absolute MeOH, and dilutions prepared in pH 8.0 phosphate buffer. Roxarsone was tested at 10, 25 and 50 mcg/ml, and salinomycin at 1, 2.5, 5.0, 10 and 25 mcg/ml. Combinations of lincomycin at 1.0 mcg/ml plus roxarsone at 50 mcg/ml and salinomycin at the same concentrations as above were also tested. Recovery of lincomycin was not interfered with as judged be zone size, and lincomycin recovery was excellent.

    To determine the recovery of arsenic due to roxarsone in the presence of salinomycin and lincomycin, each antibiotic was added to liver tissue at the 100 ppm level. Roxarsone was added to liver tissue at the 3.5 ppm level, which is the equivalent to 1.00 ppm arsenic (roxarsone is 28.48% arsenic). The antibiotics did not produce high blank nor did their presence affect the arsenic recovery.

  2. The sponsor conducted two residue studies in support of this application. The first study was conducted under Protocol 84-132 at the A.H. Robins Farm in Ashland, VA. A total of 52 birds served as controls (no drug), and 52 broilers received the combination of 75.3 g/ton (0.0083%) salinomycin plus 45.4 g/ton (0.005%) roxarsone plus 4.0 g/ton (0.00044%) lincomycin from day 0 to day 41. On day 41 all birds were placed on nonmedicated feed. An equal number of male and female birds were used. Edible tissues including liver, kidney, skin/fat, and muscle were analyzed for lincomycin at 0 day withdrawal. Liver tissue was analyzed for arsenic (roxarsone) at 0, 3 and 5 day withdrawal. Skin/fat was assayed for salinomycin at 0 day withdrawal. Six birds (three male and three female) were sacrificed and assayed for salinomycin, and 12 birds (six male and six female) were sacrificed for lincomycin and roxarsone assay.

The data from this study are summarized in the following table:

Residue Depletion - Assay Results

Withdrawal Day Tissue Parameter Assay Kidney Liver* Skin/fat** Muscle

SalinomycinPermitted Upper ppb Limit - - 200 - Salinomycin Assay 0 - - 179 - Roxarsone ppm Tolerance - 2.0 - - Roxarsone Assay 0 - 1.21-5.02 - - (range of assay for 3 - 0.51-1.28 - - all broilers assayed) 5 - 0.35-0.56 - - Lincomycin ppm Tolerance 0.1 0.1 0.1 0.1 Lincomycin Assay 0 BDL BDL BDL BDL

Sensitivity of the Method: lincomycin, 0.1 ppm. BDL = Below detectable limits; no zone found. ** = The target tissue for salinomycin is skin/fat. * = The target tissue for roxarsone (arsenic) is the liver.

In a second study (86-059) conducted at A.H. Robins, a group of 10 chickens received the combination of 75.3 g/ton (0.00083%) salinomycin plus 45.4 g/on (0.005%) roxarsone plus 4.0 g/ton (0.00044%) lincomycin from day 12 to day 32. From day 32 to day 36 the chickens received roxarsone and lincomycin at the above levels in the feed and were dosed with 14-C-salinomycin equivalent to 0.0083%. The birds were sacrificed at 6 hours of withdrawal. Salinomycin assayed at 190 ppb in skin/fat by HPLC, which is below the permitted upper limit of 200 ppb.

These data support a five day withdrawal time for the salinomycin-roxarsone-lincomycin combination.

A regulatory analytical methodology for salinomycin is not required. Practical analytical methods for the determination of tissue residues of roxarsone and lincomycin 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 Act and demonstrate that salinomycin (40-60 g/ton) plus roxarsone (45.4 g/ton) plus lincomycin (2 g/ton) are safe and effective for the claims indicated in Section 2 of this FOI Summary.

This original NADA is regarded as a Category 2 application under CVM's supplemental policy, (42 FR 64367; December 23, 1977), and which did not require reevaluation of safety and efficacy data in the parent NADAs. The drugs are to be fed in Type C Medicated feeds, in accordance with Section 2 and 3 of the FOI Summary and the Blue Bird labeling attached to this document.

Residue depletion studies in this application demonstrate that residues of parent salinomycin are below 200 ppb, the permitted upper limit of parent salinomycin in skin/fat. When the level of 200 ppb in skin/fat is not exceeded, total residues of salinomycin in all edible tissues will be below their respective safe concentrations. Residues of lincomycin in edible tissues are below the tolerance of 0.1 ppm, established in 21 CFR 556.360. Residues of arsenic (from roxarsone) in edible tissues are below the tolerances of 2 ppm in uncooked edible by products (by products include liver) and 0.5 ppm in uncooked muscle tissue (21 CFR 556.360). Adequate information was submitted to demonstrate noninterference between the assays for each drug. The approval of this application will not significantly increase human exposure to drug residues.

Noninterference studies demonstrate that salinomycin plus roxarsone in the presence of lincomycin prevented an outbreak of coccidiosis when the birds were exposed to the six major species of Eimeria. The data from six well controlled floor pen studies demonstrate the effectiveness of lincomycin for improved feed efficiency in broiler chickens in the presence of salinomycin and roxarsone. CVM's guideline for drug combinations for use in animals provides for the granting of range approval for salinomycin (40-60 g/ton) plus roxarsone (45.4 g/ton) for the prevention of coccidiosis caused by Eimeria acervulina, E. brunetti, E. maxima, E. mivati, E. necatrix and E. tenella including some field strains of E. tenella that are more susceptible to roxarsone combined with salinomycin than to salinomycin alone, and for lincomycin (2 g/ton) for improved feed efficiency in broiler chickens.

Copies of applicable labels may be obtained by writing to the:

Freedom of Information Office
Center for Veterinary Medicine, FDA
7500 Standish Place
Rockville, MD 20855