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U.S. Department of Health and Human Services

Animal & Veterinary

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NADA 141-017 Saraflox® WSP - original approval

Date of Approval: August 18, 1995

I. GENERAL INFORMATION

NADA Number: 141-017

Sponsor:

Abbott Laboratories
1401 Sheridan Road
North Chicago, IL 60064

Accepted Name: sarafloxacin hydrochloride

Trade Name: SARAFLOX® WSP

Marketing Status: A prescription (Rx) product which carries the following caution statement "Federal (USA) law restricts this drug to use by or on the order of a licensed veterinarian."

 

II. INDICATIONS FOR USE

SARAFLOX® WSP is indicated in growing broiler chickens and growing turkeys for the control of mortality associated with E. coli organisms sensitive to sarafloxacin hydrochloride.

 

III. DOSAGE FORM, ROUTE OF ADMINISTRATION, AND RECOMMENDED DOSAGE

A. Dosage Form

SARAFLOX® WSP is a water soluble powder available in 145 gram packets containing 14.5 grams of sarafloxacin.

B. Route of Administration

Chickens: Administer by mixing in drinking water of growing broiler chickens at a level of 20 to 40 ppm sarafloxacin according to the following table. When using a proportioner, mix according to the following table and meter at 1 oz/gallon.

Turkeys: Administer by mixing in drinking water of growing turkeys at a level of 30 to 50 ppm sarafloxacin according to the following table. When using a proportioner, mix according to the following table and meter at 1 oz/gallon.

                                Mix Tank                 Proportioner           
                            Gal Drinking Water         Gal Stock Solution 
       Dose (ppm)           Treated per Pouch          Treated per Pouch
          20                       192                        1.5                         
          30                       128                        1.0                         
          40                        96                        0.75                        
          50                        77                        0.60                

Recommended Dosage

Water containing SARAFLOX® WSP at a level of 20 to 40 ppm for growing broiler chickens or 30 to 50 ppm for growing turkeys should be offered for ad libitum consumption as the only source of drinking water for 5 consecutive days.

 

IV. EFFECTIVENESS

In the following studies, three different powder formulations were used. Each contained the identical amount of sarafloxacin as the hydrochloride salt. The "non-formulated" powder consisted of the salt without any additional inactive ingredients. The two "formulated" powders differed with respect to the nature and amount of the inactive ingredients (stability of all three formulations was confirmed by in vitro stability test procedures). Since all three powders were water soluble and were administered as dissolved drug, there was no need to test for potential differences in the rate and extent of in vivo dissolution. However, compositional differences could alter sarafloxacin bioavailability if a component of the powder altered the flux of the drug across the intestinal wall. Such differences could occur if active transport processes were involved. Therefore, an in situ study was conducted utilizing the rat jejunum as a model for in vivo absorption. By demonstrating the linearity in the absorption process over a wide range of sarafloxacin concentrations, this study confirmed that sarafloxacin is absorbed solely by passive processes. Therefore, differences in the powder compositions were determined to be acceptable and the formulations were found to be equivalent.

A. Pivotal Broiler Studies

  1. Dose Titration Study

    1. Type of Study: This was a dose titration study in broiler chickens with an artificially-induced E. coli infection.
    2. Investigator:
      Jeffrey Davidson
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to determine the efficacy of graded doses and select an effective low end dose of sarafloxacin for the control of colibacillosis in broilers with artificially-induced disease.
      2. Animals: In this study, commercial Arbor Acres x Peterson broilers, 21 days of age; 360 males and 360 females (2 pens of 30 males and 2 pens of 30 females per treatment) were used.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture for E. coli from air sac.
      5. Dosage Form: Dosage was a hydrochloride salt containing 826 mcg of active sarafloxacin free-base per mg of hydrochloride salt.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: The doses tested were placebo, 10, 20, 30, and 40 ppm administered in drinking water for five consecutive days.
      8. Test Duration: The study spanned ten days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was mortality. Air sac lesion scores and culture results were measured. Lesions were scored according to the following system: 0 = no lesions, 1 = minimal air sac lesions, 2 = moderate air sac and pericardial lesions, and 3 = extensive air sac and pericardial lesions.
    4. Results:

      Significant decreases in mortality were seen in all treated groups as compared to unmedicated controls. Mortality was lowest at a dose of 30 ppm, but this was not significantly better than the 20 ppm dose, so the 20 ppm dose was selected for the low end of the dose range.

      Table 4.1. Mortality, Mean Air Sac Scores, and Culture Results of Broilers That Died After Challenge with E. coli

      Culture Mortality Mean Air Sac [#Positive/ Treatment Group [(Died/Total (%)] Scores Total]

      Unmedicated/Uninfected 2/120 (0.8%) 0.00 2/2 Unmedicated/Infected 61/120 (50.8%) 2.69 61/61 Treated 10 ppm 6/120 (5.0%) 2.06 6/6 Treated 20 ppm 3/120 (2.5%) 2.25 3/3 Treated 30 ppm 1/120 (0.8%) 2.00 1/1 Treated 40 ppm 2/120 (1.7%) 2.50 2/2

    5. Statistical Analysis: A randomized complete block design was used with treatment randomly assigned within each of four blocks. Since there was little or no mortality among the sarafloxacin-treated groups, none of the data were analyzed statistically.
    6. Conclusion:

      Sarafloxacin was effective in controlling mortality associated with an artificially-induced E. coli infection. Based on a reduction in mortality, sarafloxacin at a dose of 20 ppm was determined to be the lower end of the dose range.

    7. Adverse Reactions: No adverse reactions were observed.
  2. Dose Confirmation Study

    1. Type of Study: A dose confirmation study was conducted using broiler chickens with an artificially-induced E. coli infection.
    2. Investigator:
      Diane Fagerberg, Ph.D.
      Colorado Animal Research Enterprises, Inc.
      Fort Collins, CO 80524
    3. General Design:

      1. Purpose: The purpose of this study was to confirm the dose of 20 ppm sarafloxacin for the control of colibacillosis in broilers with artificially-induced disease.
      2. Animals: In this study, commercial Peterson x Hubbard broilers, 9 days of age; 180 males and 180 females (2 pens of 30 males and 2 pens of 30 females) per treatment were used.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture of E. coli from liver and air sac.
      5. Dosage Form: Dosage was a hydrochloride salt containing 800 mcg of active sarafloxacin free-base per mg of hydrochloride salt.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo and 20 ppm administered in the drinking water for five consecutive days.
      8. Test Duration: The study spanned ten days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was mortality attributed to E. coli. Culture of air sac, pericardial, and perihepatic tissue and lesion scores were measured.
    4. Results:

      In this study, mortality was significantly reduced by treatment with sarafloxacin as opposed to the untreated control group. 98% of the untreated mortalities had positive cultures for E. coli. The treated mortalities had a much lower percentage of positive cultures.

      Table 4.2. Mortality

      Group Males Deaths Females Deaths All Deaths /Total (%) /Total (%) /Total (%)

      Non-challenged 1/60 (2%) 1/60 (2%) 2/120 (2%) Non-medicated 23/60 (38%) 23/60 (38%) 46/120 (38%) Medicated (Sarafloxacin) 3/60 (5%) 0/60 (0%) 3/120 (2%)

      *Non-medicated group vs. medicated group (p<0.05)

      Table 4.3. E. coli Culture Results 

      Total Positive Positive Culture for E. coli Culture (%)

      Group Dead (%) Survivors (%) Results Non-challenged 1/2 (50%) 8/11 (73%) 9/13 (69%) Non-medicated 45/46 (98%) 18/74 (24%) 63/120 (53%) Medicated (Sarafloxacin) 1/3 (33%) 9/117 (8%) 10/120 (8%)

      *Non-medicated vs. medicated group (p<0.05)

    5. Statistical Analysis: A randomized complete block design was used with random assignment of treatment within each block. Main effect differences were statistically evaluated by analysis of variance procedures appropriate to a randomized block design. The least-significant-difference (LSD) method was used for pair-wise comparisons of treatment means. Tests for significant difference were p< or =0.05.
    6. Conclusion: Sarafloxacin, at a dose of 20 ppm was confirmed to be effective in controlling mortality associated with an artificially-induced E. coli infection.
    7. Adverse Reactions: No adverse reactions were observed.
  3. Dose Confirmation Study

    1. Type of Study: A dose confirmation study was conducted in broiler chickens with an artificially-induced E. coli infection.
    2. Investigator:
      Diane Fagerberg, Ph.D.
      Colorado Animal Research Enterprises, Inc.
      Fort Collins, CO 80524
    3. General Design:

      1. Purpose: The purpose of this study was to confirm the dose of 20 ppm sarafloxacin for the control of colibacillosis in broilers with artificially-induced disease. Also, two formulations of sarafloxacin were compared.
      2. Animals: Commercial Arbor Acre x Arbor Acre broilers, 20 days of age; 240 males and 240 females (2 pens of 30 males and 2 pens of 30 females per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture of E. coli from liver and air sac.
      5. Dosage Form: Dosages were a hydrochloride salt containing 800 mcg of active sarafloxacin free-base per mg of hydrochloride salt and a formulated water soluble powder containing 94.82 mg of active sarafloxacin free-base per gram of powder.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo and 20 ppm administered in the drinking water for five consecutive days.
      8. Test Duration: The study spanned ten days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was mortality attributed to E. coli. Culture of air sac, pericardial, and perihepatic tissue and lesion scores were measured.
    4. Results:

      There was a significant decrease in mortality in the medicated as opposed to the non-medicated groups. The non-formulated and the formulated sarafloxacin produced similar decreases in mortality. The lowest mortality (32%) was achieved with the formulated sarafloxacin.

      Table 4.4. Mortality

      Group Females Deaths Males Deaths All Deaths /Total (%) /Total (%) /Total (%)

      Non-challenged 2/60 (3%) 2/60 (3%) 4/120 (3%) Non-medicated 34/60 (53%) 32/60 (57%) 66/120 (55%) Sarafloxacin 18/60 (30%) 28/60 (47%) 46/120 (38%) non-formulated Sarafloxacin 17/60 (28%) 22/60 (37%) 39/120 (32%) formulated

      *Non-medicated group vs. sarafloxacin soluble powder medicated group (p<0.05)

      Table 4.5. E. coli Culture Results 

      Positive Culture for E. coli Total Positive Culture

      Group Dead (%) Survivors (%) Results (%) Non-challenged 2/4 (50%) 34/116 (29%) 36/120 (30%) Non-medicated 66/66 (100%) 31/54 (57%) 97/120 (81%) Sarafloxacin 45/46 (98%) 24/74 (32%) 69/120 (57%) non-formulated Sarafloxacin 38/39 (97%) 27/81 (8%) 65/120 (54%) formulated

      *Non-medicated vs. medicated group (p<0.01)

    5. Statistical Analysis: A randomized complete block design was used with treatment randomly assigned within each block. Main effect differences were statistically evaluated by analysis of variance procedures appropriate to a randomized block design. The least-significant-difference (LSD) method was used for pair-wise comparisons of treatment means. Tests for significant difference were p < or =0.05.
    6. Conclusion: Sarafloxacin, at a dose of 20 ppm was confirmed to be effective in controlling mortality associated with an artificially-induced E. coli infection. In addition, formulated sarafloxacin was shown to be as effective as unformulated sarafloxacin.
    7. Adverse Reactions: No adverse reactions were observed.
  4. Dose Confirmation Study

    1. Type of Study: A dose confirmation study in broiler chickens with an artificially-induced E. coli infection.
    2. Investigator:
      Jeffrey Davidson
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to confirm the dose of 20 ppm sarafloxacin for the control of colibacillosis in broilers with artificially-induced disease. Also, two formulations of sarafloxacin were compared.
      2. Animals: Commercial Peterson x Arbor Acre broilers, 21 days of age; 240 males and 240 females (2 pens of 30 males and 2 pens of 30 females per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture of E. coli from pericardium and air sac.
      5. Dosage Form: Dosages were a non-formulated hydrochloride salt containing 826 mcg of active sarafloxacin free-base per mg of hydrochloride salt and a formulated water soluble powder containing 94.82 mg of active sarafloxacin free-base per gram of powder.
      6. Route of Administration: The drug was given orally in drinking water.
      7. Doses: Doses were placebo and 20 ppm administered in the drinking water for five consecutive days.
      8. Test Duration: The study spanned ten days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was mortality. Culture of air sac, pericardial, and perihepatic tissue and lesion scores were measured.
    4. Results:

      There was a significant decrease in mortality in the medicated as opposed to the non-medicated groups. The non-formulated and the formulated sarafloxacin produced similar decreases in mortality. The lowest mortality (1%) was achieved with the formulated sarafloxacin. The results are summarized in Table 4.6 and 4.7.

      Table 4.6. Mortality Day 0 to Day 10

      Group Females Deaths Males Deaths All Deaths /Total (%) /Total (%) /Total (%)

      Non-challenged 0/60 (0%) 0/60 (0%) 0/120 (0%) Non-medicated 15/60 (25%) 17/60 (28%) 32/120 (27%) Sarafloxacin (20 2/60 (3%) 2/60 (3%) 4/120 (3%) ppm) non-formulated Sarafloxacin 0/60 (0%) 1/60 (2%) 1/120 (1%) formulated

      *Non-medicated group vs. other groups (p<0.05)

      Table 4.7. E. coli Culture Results

      Group Positive Culture for E. coli Mortalities

      Non-challenged N/A* Non-medicated 32/32 Sarafloxacin (non-formulated) 4/4 Sarafloxacin (formulated) 1/1

      *Not applicable due to no mortalities in this group

    5. Statistical Analysis: A randomized complete block design was used with treatment randomly assigned within each of four blocks. Main effect differences were statistically evaluated by analysis of variance procedures. Pair-wise comparisons of treatment means performed based on the least-significant-difference (LSD) method for those variables with overall significance of p< or = 0.05. Mortality was statistically analyzed as the observed percent in order to test for interaction and differences between the sexes.
    6. Conclusion: Sarafloxacin at 20 ppm was confirmed to be effective in controlling mortality associated with an artificially-induced E. coli infection. In addition, formulated sarafloxacin was shown to be as effective as unformulated sarafloxacin.
    7. Adverse Reactions: No adverse reactions were observed.
  5. Field Study

    1. Type of Study: A field study was done in broiler chickens with colibacillosis occurring spontaneously.
    2. Investigator:
      Michael Sims
      Virginia Scientific Research
      Harrisonburg, VA 22801
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality associated with spontaneously occurring colibacillosis in broiler chickens.
      2. Animals: In this study, commercial Ross x Arbor Acre broilers, 14 days of age, 1728 males, 1728 females (24 pens with 36 males and 36 females per pen for each treatment) were used.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and isolation of E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was a water soluble powder formulation containing 98 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses used were placebo and 20 ppm administered in the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug

        effectiveness in this study was controlling mortality due to E. coli.

    4. Results

      Table 4.8. Mortality From the Initiation of Medication to Conclusion

      Treatment Mortality Associated Total Deaths With Colibacillosis (All Causes) Deaths/Total (%) Deaths/Total (%)

      Non-medicated 14/1684 (2.40%) 52/1684 (3.10%) Medicated 9/1693 (0.50%) 19/1693 (1.10%)

      * Non-medicated group vs. medicated group (p<0.01) ** Non-medicated group vs. medicated group (p<0.05)

    5. Statistical Analysis: A randomized complete block design was used with

      12 blocks, each consisting of four pens. Data for this study were evaluated by (ANOVA) using the GLM procedures of SAS. The model included block and treatment for each variable analyzed. A survival model and weighted least squares analysis were used to examine total andE. coli mortality. Chi-square tests were employed to statistically compare the overall survival experience across levels of each main effect. Tests of significance were at p < or =0.05.

    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 20 ppm was effective in controlling E. coli-associated mortality.
    7. Adverse Reactions: No adverse reactions were observed.
  6. Field Study

    1. Type of Study: A field study in broiler chickens with spontaneously occurring colibacillosis.
    2. Investigator:
      Michael Sims
      Virginia Scientific Research
      Harrisonburg, VA 22801
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality associated with spontaneously occurring colibacillosis.
      2. Animals: Commercial Peterson x Arbor Acre broilers, 24 days of age; 2304 males (16 pens per treatment with 72 males per pen) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and isolation of E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was a water soluble powder formulation containing 98 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo and 20 ppm administered via the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality due to E. coli.
    4. Results:

      Use of sarafloxacin at 20 ppm administered in the drinking water for 5 days reduced colibacillosis associated mortality from 3.8% to .62%.

      Table 4.9. Mortality From the Initiation of Medication to Conclusion

      Treatment Mortality Associated Total Deaths With Colibacillosis (All Causes) Deaths/Total (%) Deaths/Total (%)

      Non-medicated 43/1143 (3.80%) 44/1143 (3.80%) Medicated 7/1136 (0.62%) 17/1136 (1.50%)

      * Non-medicated group vs. medicated group (p<0.01) ** Non-medicated group vs. medicated group (p<0.05)

    5. Statistical Analysis: A randomized complete block design was used with

      16 blocks, each consisting of three pens. Data for this study were evaluated by (ANOVA) using the GLM procedures of SAS. The model included block and treatment for each variable analyzed. A survival model and weighted least squares analysis were used to examine total and E. coli mortality. Chi-square tests were employed to statistically compare the overall survival experience across levels of each main effect. Tests of significance were at p < or =0.05.

    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 20 ppm was effective in controlling mortality associated with colibacillosis.
    7. Adverse Reactions: No adverse reactions were observed.
  7. Field Study

    1. Type of Study: A field study was conducted in broiler chickens with spontaneously occurring colibacillosis.
    2. Investigator: Michael Sims
      Virginia Scientific Research
      Harrisonburg, VA 22801
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality associated with spontaneously occurring colibacillosis.
      2. Animals: Commercial Peterson x Arbor Acre broilers, 36 days of age; 1728 females and 1728 males (24 pens per treatment with an equal number of males and females per pen) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and isolation of E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was a water soluble powder formulation to be marketed containing 107 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox ® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo and 20 ppm administered in drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was controlling mortality due to E. coli.
    4. Results:

      Use of sarafloxacin at 20 ppm administered in the drinking water for 5 days reduced colibacillosis associated mortality from 15.68% to 6.94%.

      Table 4.10. Mortality From the Initiation of Medication to Conclusion

      Treatment Mortality Associated Total Deaths With Colibacillosis (All Causes) Deaths/Total (%) Deaths/Total (%)

      Non-medicated 281/1792 (15.68%) 343/1792 (19.14%) Medicated 114/1643 (6.94%) 243/1643 (14.79%)

      * Non-medicated group vs. medicated group (p<0.01) ** Non-medicated group vs. medicated group (p<0.05)

      e. Statistical Analysis: A randomized complete block design was used with 12 blocks, each consisting of four pens. Data for this study were evaluated by (ANOVA) using the GLM procedures of SAS. The model included block and treatment for each variable analyzed. A survival model and weighted least squares analysis were used to examine total and E. coli mortality. Chi-square tests were employed to statistically compare the overall survival experience across levels of each main effect. Tests of significance were at p < or =0.05.

    5. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 20 ppm was effective in controlling mortality associated with colibacillosis.
    6. Adverse Reactions: No adverse reactions were observed.
  8. Field Study

    1. Type of Study: A field study was conducted in broiler chickens with spontaneously occurring colibacillosis.
    2. Investigator:
      Ed Odor
      University of Delaware Research Farm
      & Education Center
      Georgetown, DE 19947
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin in chickens for the control of a natural outbreak of colibacillosis.
      2. Animals: In this study, Peterson x Arbor Acres straight-run broilers, 36 days of age were used. Treatment groups were a non-medicated group of 1584 chickens and a sarafloxacin-medicated group of 1590 chickens (48 pens arranged in twelve blocks of four pens each). Each pen contained at least 64 birds but no more than 72 birds.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Mortality associated with colibacillosis was not considered conclusive until either samples taken from air sac, heart or liver tissues tested positive for E. coli upon culture, or lesions associated with colibacillosis were present.
      5. Dosage Form: Dosage was a water soluble powder formulation containing 98 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo and 20 ppm administered in the water supply for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality due to E. coli.
    4. Results:

      Use of sarafloxacin at 20 ppm administered in the drinking water for 5 days reduced colibacillosis associated mortality from 2.75% to 1.37%.

      Table 4.11. Mortality Results

      Group Mortality Associated Total Deaths With Colibacillosis (All Causes) Deaths/Total (%) Deaths/Total (%)

      Non-medicated 46/1584 (2.75%) 70/1584 (4.42%) Medicated 22/1590 (1.37%) 50/1590 (3.14%)

      * Non-medicated group vs. medicated group (p<0.05)

    5. Statistical Analysis: A randomized complete block design was used with

      12 blocks, each consisting of four pens, two of each treatment. Data for this study were evaluated by (ANOVA) using the GLM procedures of SAS. The model included block and treatment for each variable analyzed. A survival model and weighted least squares analysis were used to examine total and E. coli mortality. Chi-square tests were employed to statistically compare the overall survival experience across levels of each main effect. Tests of significance were at p < or =0.05.

    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at a concentration of 20 ppm was shown to be effective in controlling mortality associated with colibacillosis.
    7. Adverse Reactions: No adverse reactions were observed.
  9. Field Study

    1. Type of Study: A field study was conducted in broiler chickens for the control of a natural outbreak of colibacillosis.
    2. Investigator:
      Ed Odor
      University of Delaware Research Farm
      & Education Center
      Georgetown, DE 19947
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin in broiler chickens undergoing a natural outbreak of colibacillosis.
      2. Animals: Commercial Peterson x Arbor Acres broilers, 30 days of age; approximately 3168 straight-run broilers (24 pens per treatment with 64 to 72 birds per pen) were used for this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and isolation of E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was the water soluble powder formulation containing 98 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo and 20 ppm administered via the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality due to E. coli.
    4. Results:

      Use of sarafloxacin at 20 ppm administered in the drinking water for 5 days reduced colibacillosis associated mortality from 2.59% to .63%.

      Table 4.12. Mortality Results

      Group Mortality Associated Total Deaths With Colibacillosis (All Causes) Deaths/Total (%) Deaths/Total (%)

      Non-medicated 41/1580 (2.59%) 197/1580 (12.47%) Medicated 10/1581 (0.63%) 170/1581 (10.75%)

      *Non-medicated group vs. medicated group (p<0.01)

    5. Statistical Analysis: A randomized complete block design was used with 12 blocks, each consisting of four pens, two of each treatment. Data for this study were evaluated by (ANOVA) using the GLM procedures of SAS. The model included block and treatment for each variable analyzed. A survival model and weighted least squares analysis were used to examine total and E. coli mortality. Chi-square tests were employed to statistically compare the overall survival experience across levels of each main effect. Tests of significance were at p < or = 0.05.
    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 20 ppm was effective in controlling mortality associated with colibacillosis.
    7. Adverse Reactions: No adverse reactions were observed.
  10. Field Study

    1. Type of Study: A field study was conducted using broiler chickens with spontaneously occurring colibacillosis.
    2. Investigator:
      Jeffrey Davidson, DVM
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality associated with spontaneously occurring colibacillosis.
      2. Animals: Commercial Arbor Acre x Arbor Acre broilers, 22 days of age; 2700 straight-run broilers (27 pens per treatment with 50 birds per pen) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and isolation of E. coli from the pericardium and air sac.
      5. Dosage Form: Dosage was the water soluble powder formulation containing 98 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses were placebo and 20 ppm administered via the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality attributed to E. coli.
    4. Results:

      Use of sarafloxacin at 20 ppm administered in the drinking water for 5 days reduced colibacillosis associated mortality from 3.7% to 1.2%.

      Table 4.13. Mortality From the Initiation of Medication to Conclusion

      Treatment Mortality Associated Total Deaths With Colibacillosis (All Causes) Deaths/Total (%) Deaths/Total (%)

      Non-medicated 49/1322 (3.7%)* 52/1324 (3.9%) Medicated 16/1318 (1.2%)* 19/1320 (1.4%)

      * Results not recorded for two birds ** Non-medicated group vs. medicated group (p <0.01)

    5. Statistical Analysis: A randomized complete block design was used with 16 blocks, each consisting of three pens. Data for this study were evaluated by (ANOVA) using the GLM procedures of SAS. The model included block and treatment for each variable analyzed. A survival model and weighted least squares analysis were used to examine total and E. coli mortality. Chi-square tests were employed to statistically compare the overall survival experience across levels of each main effect. Tests of significance were at p < or =0.05.
    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 20 ppm was effective in controlling mortality associated with colibacillosis.
    7. Adverse Reactions: No adverse reactions were observed.

B. Corroborative Broiler Studies

  1. Range-Finding Study

    1. Type of Study: A range-finding study was conducted in broiler chickens with an artificially-induced E. coli infection.
    2. Investigator: Jeffrey Davidson
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to determine the efficacy of graded doses and select an appropriate low end dose of sarafloxacin for the control of colibacillosis in broilers with artificially-induced disease.
      2. Animals: Commercial Arbor Acres x Peterson broilers, 21-days of age; 360 males and 360 females (2 pens of 30 males and 2 pens of 30 females per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture of E. coli from air sac.
      5. Dosage Form: Dosage was a hydrochloride salt containing 826 mcg of active sarafloxacin free-base per mg of hydrochloride salt.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo, 12.5, 25.0, 50.0, and 75.0 ppm administered in the drinking water for five consecutive days.
      8. Test Duration: The study spanned ten days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was mortality. Air sac lesion scores and culture results were also measured.
    4. Results:

      Significant decreases in mortality were seen in all treated groups as compared to unmedicated controls. Mortality was lowest at a dose of 25 ppm and 75 ppm.

      Table 4.14. Mortality, Mean Air Sac Scores, and Culture Results of Broilers That Died After Challenge with E. coli

      Treatment Group Mortality Mean Air Culture (Died/Total %) Sac Scores (#Positive /Total)

      Unmedicated/Uninfected 1/120 (0.8%) 0 0/1 Unmedicated/Infected 36/120 (30.0%) 2.1 36/36 Treated 12.5 ppm 2/120 (1.7%) 2.0 2/2 Treated 25 ppm 0/120 (0%) 0 0/0 Treated 50 ppm 2/120 (1.7%) 1.5 2/2 Treated 75 ppm 0/120 (0%) 0 0/0

    5. Statistical Analysis: A randomized complete block design was used with treatment randomly assigned within each of four blocks. Since there was little or no mortality or morbidity in the groups that received sarafloxacin, none of the data were analyzed statistically.
    6. Conclusion: Sarafloxacin at a level of 12.5 to 75 ppm was effective in controlling mortality associated with an artificially-induced E. coli infection.
    7. Adverse Reactions: No adverse reactions were observed.
  2. Serum and Lung Levels of Sarafloxacin Following Administration in Drinking Water to Broilers

    1. Type of Study: This was a study to determine the serum and lung homogenate concentration profile for sarafloxacin.
    2. Investigator:
      Terry TerHune
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to determine the average concentration of sarafloxacin in serum and in homogenated lung samples on Day 3 of ad libitum access to medicated water as representative of concentrations expected during field use. This study was intended to support the efficacy of the doses within the proposed dose range.
      2. Animals: Healthy growing broilers from a local commercial grower, were used in this study. 300 broilers, 150 of each sex, were assigned to six pens, three pens of each sex, with 50 birds per pen on the basis of weight.
      3. Route of Administration: The drug was administered orally in drinking water.
      4. Doses: Doses tested were 20, 30, or 40 ppm administered in the drinking water for three consecutive days. Blood samples were taken before treatment and on the third day at 7, 9, and 11 a.m., and 1, 4, 7, and 10 p.m., plus at 7 a.m. the following day. Lung samples were taken from birds bled at 48, 52, 57, and 63 hours.
      5. Test Duration: The study spanned four days.
      6. Pertinent Parameters Measured: Sarafloxacin levels were measured in blood and lung homogenate samples. Water was analyzed to determine sarafloxacin levels.
    4. Results:

      Average serum and lung concentrations are presented on a sarafloxacin free base basis to be comparable to the units used for bacterial susceptibility determinations, and are adjusted to the actual dose of sarafloxacin administered as reflected by the percent sarafloxacin analyzed in the drinking water.

      Table 4.15 Drug Concentration in Serum and Lung Homogenate

      Serum Concentration Lung Homogenate Mean ± SD Concentration Dose (ppm) (µg/ml) Mean ± SD (µg/g)

      20 .052 ± .016 .086 ± .029 30 .066 ± .020 .127 ± .042 40 .083 ± .025 .175 ± .069

    5. Statistical Analysis: The adjusted serum and lung concentrations were calculated by linear regression analysis.
    6. Conclusion: The average serum concentrations exceeded the MIC50 at all doses. The serum concentrations may be more reliable than the lung concentration, as the actual availability of the drug at the target tissue is not known. If the concentration of drug should be no less than the MIC concentration, then the 20 ppm dose should be restricted to use when the pathogen causing disease has an MIC at or below the MIC50. The 30 and 40 ppm doses are expected to achieve serum and tissue average concentrations above the MIC90 and should be effective in a larger percentage of disease outbreaks.

C. Pivotal Turkey Studies

  1. Dose Determination 

    1. Type of Study: A dose determination study was conducted in young growing turkeys in which colibacillosis was artificially induced by intramuscular injection of E. coli.
    2. Investigator:
      Jeffrey Davidson
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to determine the efficacy of graded doses of sarafloxacin and select an effective low end dose for the control of mortality due to pathogenic E. coli in growing turkeys.
      2. Animals: Commercial Broad Breasted White turkeys, 11 weeks of age; 288 males and 384 females (48 pens: 6 pens of 12 males and 6 pens of 16 females per treatment) were used.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and isolation of E. coli from pericardium and air sac.
      5. Dosage Form: Dosage was the water soluble powder formulation containing 92 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses tested were placebo, 15, 30, and 45 ppm administered in the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality attributed to E. coli.
    4. Results:

      Of the 77 turkeys which died during the study, deaths due to colibacillosis were 27 in the infected, non-medicated group, compared to 18, 9, and 8 in the medicated groups receiving 15, 30, and 45 ppm sarafloxacin, respectively.

      Table 4.16. Mortality From the Initiation of Medication to Study End

      Treatment Group Death Due to E. coli* Total Mortality Deaths/Total (%) Deaths/Total (%)

      Non-medicated 27/168 (16.1%) 34/168 (20.2%) 15 ppm sarafloxacin 18/168 (10.7%) 23/168 (13.7%) 30 ppm sarafloxacin 9/168 (5.4%) 10/168 (6.0%) 45 ppm sarafloxacin 8/168 (4.8%) 10/168 (6.0%)

      * Non-medicated group vs. medicated group (p<0.05)

      A poult death was counted as having been due to colibacillosis only if:

      a) air sac or pericardial tissue cultured positive for E. coli,
      b) the colibacillosis lesion score was greater than 1, and
      c) death occurred after treatment was initiated.

    5. Statistical Analysis: Stepwise single-degree-of-freedom non-orthogonal contrasts were examined for the main effect of the treatment. Each of these analyses included univariate models of block, treatment, sex, and the treatment x sex interaction. A Weighted Least Squares (WLS) was used to examine the percentage of dead birds at study completion. Tests of significance were made at the p < or =0.05 level.

      Table 4.17. Analysis of Dose Level Effects on Mortality Due to Colibacillosis.

      Treatment Comparisons P-Value

      All treatments 0.007 Control vs. Treated 0.003 Control vs. 15 ppm 0.152 Control vs. 30 ppm 0.004 Control vs. 45 ppm 0.002 15 ppm vs. 30 ppm 0.082 15 ppm vs. 45 ppm 0.042 30 ppm vs. 45 ppm 0.722

    6. Conclusion: Sarafloxacin administered in drinking water at 30 ppm for five consecutive days was determined to be the low end of the dose range for reducing mortality due to an artificially-induced E. coli infection in growing turkeys.
    7. Adverse Reactions: No adverse reactions were observed.
  2. Field Study

    1. Type of Study: A field trial was conducted using growing turkeys to determine the effectiveness of sarafloxacin for the control of mortality associated with E. coli in a spontaneously occurring outbreak of colibacillosis.
    2. Investigator:
      Jeffrey Davidson
      Health Management Services
      Tulare, CA 93275
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality in an outbreak of respiratory disease due to E. coli (colibacillosis) in growing turkeys.
      2. Animals: Commercial Broad Breasted White turkeys 8 weeks of age; 288 males and 384 females (48 pens - 12 pens of 12 males and 12 pens of 16 females per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed based on examination of lesions on necropsy and culturing pericardium and air sac for E. coli.
      5. Dosage Form: Dosage was a water soluble powder formulation containing 102 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses were placebo and 30 ppm administered in drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality attributed to E. coli.
    4. Results:

      During the five-day medication and 10-day post-medication period deaths due to E. coli were 21/324 (6.5%) of the non-medicated poults (p <0.05) compared to 2/326 (0.1%) of those poults medicated with 30 ppm sarafloxacin.

      Table 4.18. Mortality From the Initiation of Medication to Study End

      Group Mortality Associated Total Deaths* with Colibacillosis* Deaths/Total (%) Deaths/Total (%)

      Non-medicated 21/324 (6.5%) 24/324 (7.41%) Medicated 2/326 (0.06%) 5/326 (1.53%)

      *Non-medicated group vs. medicated group (p<0.05)

      A poult death was counted as having been due to colibacillosis only if:

      a) air sac or pericardial tissue cultured positive for E. coli,
      b) the colibacillosis lesion score was greater than 1, and
      c) death occurred after treatment was initiated.

    5. Statistical Analysis: A randomized complete block design was used with 12 blocks, each consisting of four pens, two of each treatment. The model included block, sex, treatment, and the sex x treatment interaction for each of the variables analyzed. A Weighted Least Squares (WLS) analysis was used to examine the percentage of dead birds at study completion.
    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 30 ppm was effective in controlling mortality associated with a natural outbreak of colibacillosis in growing turkeys.
    7. Adverse Reactions: No adverse reactions were observed.
  3. Field Study

    1. Type of Study: A field trial was conducted using growing turkeys with spontaneously occurring disease to determine if sarafloxacin was effective for the control of mortality associated with E. coli in an outbreak of colibacillosis.
    2. Investigator:
      James McNaughton
      PARC Institute
      Easton, Maryland 21601
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality in an outbreak of respiratory disease due to E. coli (colibacillosis)in growing turkeys.
      2. Animals: British United Turkeys, 33 days of age (1440 females: 32 pens-16 pens of 45 females per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture for E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was a water soluble powder formulation containing 102 mg of active sarafloxacin free-base per gram of powder.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses were placebo and 30 ppm administered via the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was controlling mortality associated with E. coli.
    4. Results:

      During the five-day medication and 10-day post-medication period deaths associated with colibacillosis were 119/705 (16.9%) for the non-medicated poults compared to 62/710 (8.7%) for the sarafloxacin treated group.

      Table 4.19 Mortality From the Initiation of Medication to Study End

      Group Mortality Associated Total Mortality* with Colibacillosis* Deaths/Total (%) Deaths/Total (%)

      Non-medicated 119/705 (16.9%) 133/705 (18.9%) Medicated 62/710 (8.7%) 67/710 (9.4%)

      * Non-medicated group vs. medicated group (p<0.05)

      A poult death was counted as having been due to colibacillosis only if:

      a) air sac or pericardial tissue cultured positive for E. coli,
      b) the colibacillosis lesion score was greater than 1, and
      c) death occurred after treatment was initiated.

    5. Statistical Analysis: A randomized complete block design was used with 8 blocks each consisting of four pens. The model included block and treatment for each of the variables analyzed. A Weighted Least Squares (WLS) analysis was used to examine the percentage of dead birds at study completion.
    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 30 ppm was effective in controlling mortality associated with a natural outbreak of colibacillosis in growing turkeys.
    7. Adverse Reactions: No adverse reactions were observed.
  4. Field Study

    1. Type of Study: A field trial was conducted using growing turkeys with spontaneously occurring disease to determine if sarafloxacin was effective for the control of mortality associated with E. coli in an outbreak of colibacillosis.
    2. Investigator:
      Carey Quarles
      Colorado Quality Research
      Fort Collins, CO 80524
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality in an outbreak of respiratory disease due to E. coli (colibacillosis) in growing turkeys.
      2. Animals: Commercial Nicholas Cross Market turkeys, 18 days of age, 1200 males (40 pens-20 pens of 30 males per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culture of E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was the water soluble powder formulation containing 102 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses were placebo and 30 ppm administered via the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality attributed to E. coli.
    4. Results:

      During the five-day medication and 10-day post-medication period a total of 40/565 (7.1%) of the non-medicated poults (p<0.05) compared to 10/568 (1.8%) of those medicated with 30 ppm sarafloxacin died due to colibacillosis.

      Table 4.20 Mortality From the Initiation of Medication to Study End

      Group Mortality Associated Total Mortality* with Colibacillosis* Deaths/Total (%) Deaths/Total (%)

      Non-medicated 40/565 (7.1%) 56/565 (9.9%) Medicated 10/568 (1.8%) 17/568 (3.0%)

      * Non-medicated group vs. medicated group (p<0.05)

      A poult death was counted as having been due to colibacillosis only if:

      a) air sac or pericardial tissue cultured positive for E. coli,
      b) the colibacillosis lesion score was greater than 1, and
      c) death occurred after treatment was initiated.

    5. Statistical Analysis: A randomized complete block design was used with 10 blocks each consisting of four pens, two of each treatment. The model included block and treatment for each of the variables analyzed. A Weighted Least Squares (WLS) analysis was used to examine the percentage of dead birds at study completion.
    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 30 ppm was effective in controlling mortality associated with a natural outbreak of colibacillosis in growing turkeys.
    7. Adverse Reactions: No adverse reactions were observed.

D. Corroborative Turkey Studies

  1. Field Study

    1. Type of Study: A field trial was conducted using growing turkeys, obtained from a commercial production facility and undergoing a field outbreak of colibacillosis.
    2. Investigator:
      Michael Sims
      Virginia Scientific Research
      Harrisonburg, VA 22801
    3. General Design:

      1. Purpose: The purpose of this study was to evaluate the efficacy of sarafloxacin for the control of mortality in an outbreak of respiratory disease due to E. coli (colibacillosis) in growing turkeys.
      2. Animals: British United Turkeys #6; 56 days of age; 1056 females (48 pens - 24 pens of 22 females per treatment) were used in this study.
      3. Control: Non-medicated water was available ad libitum from waterers for the duration of the study.
      4. Diagnosis: Colibacillosis was diagnosed by necropsy and culturing for E. coli from liver, pericardium, and air sac.
      5. Dosage Form: Dosage was a water soluble powder formulation containing 99 mg of active sarafloxacin free-base per gram of powder. This dosage is the marketed form of SaraFlox® WSP.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Doses: Doses were 30 ppm administered via the drinking water for five consecutive days.
      8. Test Duration: The study spanned fifteen days.
      9. Pertinent Parameters Measured: The primary parameter for determining drug effectiveness in this study was reduction in mortality attributed to E. coli.
    4. Results:

      During the five-day medication and 10-day post-medication period a total of 19/517 (3.7%) of the non-medicated poults died compared to 11/517 (2.1%) of those medicated with 30 ppm sarafloxacin.

      Table 4.21 Mortality From the Initiation of Medication to Study End

      Group Mortality Associated Total Mortality* with Colibacillosis* Deaths/Total (%) Deaths/Total (%)

      Non-medicated 19/517 (3.7%) 31/517 (6.0%) Medicated 11/517 (2.1%) 24/517 (4.6%)

      * Non-medicated group vs. medicated group (p = 0.23)

      A poult death was counted as having been due to colibacillosis only if:

      a) air sac or pericardial tissue cultured positive for E. coli,
      b) the colibacillosis lesion score was greater than 1, and
      c) death occurred after treatment was initiated.

    5. Statistical Analysis: A randomized complete block design was used with 12 blocks each consisting of four pens, two of each treatment. The model included block and treatment for each of the variables analyzed. A Weighted Least Squares (WLS) analysis was used to examine the percentage of dead birds at study completion.
    6. Conclusion: Sarafloxacin administered in drinking water for five consecutive days at 30 ppm was effective in controlling mortality associated with a natural outbreak of colibacillosis in growing turkeys but this effect was not statistically significant.
    7. Adverse Reactions: No adverse reactions were observed.
  2. Serum and Lung Levels of Sarafloxacin Following Administration in Drinking Water to Growing Turkeys

    1. Type of Study: This was a study to determine the serum and lung homogenate concentrations profile for sarafloxacin.
    2. Investigator:
      Jonathan Schaeffer
      JLS Research
      Mount Olive, NC 28365
    3. General Design:

      1. Purpose: The purpose of this study was to determine the average concentration of sarafloxacin in serum and in homogenated lung samples on Day 3 of ad libitum access to medicated water as representative of concentrations expected during field use. This study was intended to support the efficacy of the doses within the proposed dose range.
      2. Animals: Healthy growing turkeys from a local commercial grower, were used in this study. 396 turkeys, 180 toms and 216 hens, were assigned to twelve pens, six pens of each sex, with 30 toms or 36 hens per pen on the basis of weight.
      3. Route of Administration: The drug was administered orally in drinking water.
      4. Doses: Doses tested were 30, 40, or 50 ppm administered in the drinking water for three consecutive days prior to taking a series of blood samples. Blood samples were taken before treatment and on the third day at 8, 10 a.m., and 12 p.m., and 2, 5 ,8 ,11 p.m., plus at 8 a.m. the following day. Lung samples were taken from birds bled at 48, 52, 57, and 63 hours.
      5. Test Duration: The study spanned four days.
      6. Pertinent Parameters Measured: Sarafloxacin levels were measured in blood and lung homogenate samples. Water was analyzed to determine sarafloxacin levels.
    4. Results:

      Average serum and lung concentrations are presented on a sarafloxacin free base basis to be comparable to the units used for bacterial susceptibility determinations, and are adjusted to the actual dose of sarafloxacin administered as reflected by the percent sarafloxacin analyzed in the drinking water.

      Table 4.22 Drug Concentration in Serum and Lung Homogenate

      Dose (ppm) Serum Lung Homogenate Concentration Concentration Mean ± SD (µg/ml) Mean ± SD (µg/g) 30 .021 ± .012 .069 ± .024 40 .025 ± .010 .081 ± .036 50 .041 ± .020 .126 ± .052

    5. Statistical Analysis: The adjusted serum and lung concentrations were calculated by linear regression analysis.
    6. Conclusion: The average lung concentrations exceeded the MIC90 at all doses. The serum concentrations may be more reliable than the lung concentration, as the actual availability of the drug at the target tissue is not known. If the concentration of drug should be no less than the MIC concentration, then the 30 ppm dose should be restricted to use when the pathogen causing disease has an MIC at or below the MIC50. The 40 and 50 ppm doses are expected to achieve tissue average concentrations above the MIC90 and should be effective in a larger percentage of disease outbreaks.

 

V. ANIMAL SAFETY SUMMARY

A. Growing Broilers

  1. Safety Study

    A specifically designed Target Animal Safety Study in broiler chickens was conducted to address the tolerance to and safety of sarafloxacin. Based on the results of this study, sarafloxacin is safe when administered in the drinking water up to 153 ppm for 15 days.

    1. Type of Study: This was a 22-day study in broilers in which sarafloxacin was administered in the drinking water at 0, 15, 46 and 153 ppm for 15 consecutive days followed by a seven-day observation period.
    2. Study Director:
      Jeffrey Davidson, DVM, MPVM
      Health Management Services
      Tulare, CA 93274
    3. General Design:

      1. Purpose: To determine the toxicologic effects of sarafloxacin when administered in the drinking water. Potential target organs and tissues were to be identified through clinical observations, gross necropsy and histologic examination. Red blood cell count, hemoglobin, packed cell volume, clotting time, body weight, feed and water consumption were other variables of interest.
      2. Animals: 320 day-old commercial strain broilers (160 males, 160 females), 40 males and 40 females/treatment: 2 pens of each gender; were used in this study. Their average initial weight was 42.4 grams.
      3. Control: Non-medicated drinking water was available ad libitum.
      4. Dosage form: A water soluble powder containing 94 mg of active sarafloxacin free-base per gram of powder was used. This dosage is the marketed form of SaraFlox® WSP.
      5. Dosage used: The doses were 0, 15, 46 and 153 ppm sarafloxacin.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Study Duration: This study included 15 consecutive days of medication followed by 7 days of observation.
      8. Pertinent measurements/observations: Clinical observations, hematologic measurements, body weight changes, feed and water consumption, gross necropsy and histopathology were evaluated to assess the potential toxicity in broilers.
    4. Results:

      1. Clinical Observations:

        No clinical signs of toxicity or illness were observed during this study. Droppings, feathering and behavior were normal throughout the 22-day study period. No deaths occurred during the study.

      2. Water Consumption:

        Sarafloxacin had no effect on water consumption. Minor variances in intake resulted in day x sex interactions which were of no clinical concern.

      3. Body Weight:

        At the end of the 15-day period of medication, birds at 15 ppm sarafloxacin were heavier (P<0.01) than the non-medicated controls. At Day 22, birds in the 15 ppm group were heavier (p<0.05) than those in the 46 ppm group. Males were heavier (p<0.01) than females at day 15 and 22.

      4. Feed Consumption:

        Feed consumption was influenced by gender with males consuming more than females (p<0.01). Birds at 15 ppm sarafloxacin consumed more feed (p<0.05) than each of the other groups for the study period.

      5. Hematology:

        Slight, but significant reductions in packed cell volume occurred at Day 4 in the 15 ppm treatment group compared with the control group (p<0.05) and the higher doses of sarafloxacin (p<0.01). Clotting time at Day 4 was less (P<0.01) among chicks in the 15 ppm group than in the 46 ppm group. However, it should be noted that all mean hematological values were within reported ranges for chickens.

      6. Gross and Histological Observations:

        Gross lesions were found among all treatment groups and generally described as mild and not dose related. Typically, airsacculitis, pericarditis, enteritis and atrophy of the bursa were observed.

        Histopathological changes in both the 0 ppm and 153 ppm treatment groups were generally confined to the liver, kidney or heart except for an occasional occurrence in the brain and gonads of one or two birds in each group. These lesions were multifocal, minimal lymphoid reactions and (or) extramedullary hematopoiesis, primarily in the heart and considered to be common in young birds.

        Body weight and feed consumption effects were generally positive, but not dose related. Mean hematology values were within reported ranges for chickens. Gross and microscopic tissue changes were mild and not dose-related.

      Table 5.1 Frequencies of Various Findings

      Tissue/Microscopic Observations Treatment

      0 ppm 153 ppmLiver No lesion recognized 7/20 1/19 (1NE)(a) Lymphoid nodules 12/20 15/19 Extramedullary hematopoiesis 1/20 3/19 Kidney No lesion recognized 14/20 15/20 Lymphoid nodules 6/20 5/20 Heart No lesion recognized 15/20 7/20 Lymphoid nodules 0/20 8/20 Extramedullary hematopoiesis 5/20 5/20 Epicarditis 0/20 1/20 Gonads No lesion recognized 16/18 (2NE)* 19/20 Lymphoid nodules 2/18 1/20 Extramedullary hematopoiesis 0/18 1/20

      * a NE = not examined

    5. Statistical Analysis:

      A randomized complete block design was used with four treatments in four replicate blocks. The model contained the main effects of location (i.e., row), sex, treatment and the sex x treatment interaction. Day was included in the model for water consumption, a variable with multiple measurements over time. The experimental unit for weight, feed consumption and water intake was the pen of animals; whereas, individual birds were considered as independent experimental units for each hematological variable.

      Single-degree-of freedom contrasts were used to measure treatment differences. These a priori contrasts were 0 ppm vs. 15 ppm; 15 ppm vs. 46 ppm and 15 ppm vs. 153 ppm sarafloxacin.

    6. Conclusion:

      Treatment related effects were biologically insignificant when sarafloxacin was administered in the drinking water for 15 days in broiler chicks at levels of 15 to 153 ppm. There were no clinical signs of toxicity or illness among any of the groups. Therefore the drug was demonstrated to be safe at 3.8X the maximum recommended dose and 7.7X the minimum recommended dose for 3X the recommended duration of treatment.

B. Growing Turkeys

A specifically designed target animal safety study with turkey poults was conducted to address the tolerance to and safety of sarafloxacin. Based on the results of this study, sarafloxacin is safe when administered in the drinking water up to at least 300 ppm for 15 days.

  1. Safety Study

    1. Type of Study: This was a 22-day study in turkeys in which sarafloxacin was administered in the drinking water at 0, 30, 90 and 300 ppm for 15 consecutive days followed by a seven-day observation period.
    2. Study Director:
      Jeffrey Davidson, DVM, MPVM
      Health Management Services, Inc.
      Tulare, CA 93274
    3. General Design:

      1. Purpose: This study was conducted to determine the toxicologic effects of sarafloxacin in turkey poults when administered in the drinking water. Potential target organs and tissues were to be identified through clinical observations, gross necropsy, and histologic examination. Red blood cell count, hemoglobin, packed cell volume, clotting time, body weight, feed and water consumption were other variables of interest.
      2. Animals: 320 eight-week old Nicholas Broad-Breasted White Turkeys, 160 males, 160 females, 40 males and 40 females per treatment (2 pens of each gender) were used in this study. The average initial weight for females was 4.77 kg, and for males was 5.66 kg.
      3. Control: Non-medicated drinking water was available ad libitum.
      4. Dosage Form: A water soluble powder containing 92 mg of active sarafloxacin free-base per gram of powder was used. This dosage is the marketed form of SaraFlox® WSP.
      5. Dosage used: The doses used were 0, 30, 90 and 300 ppm of sarafloxacin.
      6. Route of Administration: The drug was administered orally in drinking water.
      7. Study Duration: The study included fifteen consecutive days of medication beginning at nine weeks of age followed by

        7 days of observation.

      8. Pertinent measurements/observations: Clinical observations, hematologic measurements, body weight changes, feed and water consumption, gross necropsy and histopathology were evaluated to assess the potential toxicity in broilers.
    4. Results:

      1. Clinical Observations:

        No signs of clinical toxicity or illness were observed during this study. Droppings, feathering and behavior were normal throughout the 22-day study period. None of the poults died during the study.

      2. Water Consumption:

        Water intake was significantly reduced (P<0.01) at 300 ppm sarafloxacin throughout the medication period; however, intake was similar for all groups during the post-medication period. Males consumed more than females.

      3. Body Weight:

        The only effect of sarafloxacin on body weight occurred at day 7 of medication when poults at 30 ppm were heavier (p<0.01) than the 300 ppm group. Males were heavier than females (P<0.01) throughout the study.

      4. Feed Consumption:

        Feed consumption was influenced by gender with the heavier males consuming more than females (P<0.01). Birds at 300 ppm sarafloxacin tended to consume less feed than the other groups during the first four days of medication.

      5. Hematology:

        No significant statistical differences in hematology values were observed among the study groups and all values were within the normal clinical range for turkeys.

      6. Gross and Histological Observations:

        Gross lesions were found among all treatment groups and generally described as mild or small. Minimal lesions of fowl pox, typical in poults this age, were seen in all groups. Airsacculitis and enteritis were also found in each group.

        Histopathological changes at both the 0 ppm and 300 ppm treatment levels were incidental and minimal in both groups. Microscopic review of liver and kidney tissues showed multifocal, minimal lymphoid nodules, in the majority of poults examined in both groups. Cardiac tissue from half the poults in both groups showed evidence of extramedullary hematopoiesis, congestion and (or) hemorrhage. Lymphoid nodules were seen in gonadal tissue in 2/20 poults in the control group and in 8/20 in the 300 ppm sarafloxacin group.

      Table 5.2 Frequencies of Various Findings

      Tissue/Microscopic Observations Treatment

      0 ppm 300 ppmLiver No lesion recognized 7/20 7/20 Lymphoid nodules 13/20 13/20 Hepatocyte vacuolation 0/20 1/20 Kidney No lesion recognized 6/19 (1NE*) 7/20 Lymphoid nodules 13/19 13/20 Heart No lesion recognized 10/20 10/20 Lymphoid nodules 3/20 2/20 Extramedullary hematopoiesis 1/20 0/20 Congestion 5/20 6/20 Hemorrhage 1/20 0/20 Congestion & Hemorrhage 1/20 1/20 Gonads No lesion recognized 17/20 12/20 Lymphoid nodules 3/20 3/20

      * NE = not examined

    5. Statistical Analysis:

      A randomized complete block design was used with four treatments in four replicate blocks. The model contained the main effects of location (i.e. row), sex, treatment and the sex x treatment interaction. Day was included in the model for water consumption, a variable with multiple measurements over time.

      The experimental unit for weight, feed consumption and water intake was the pen of animals; whereas, individual birds were considered as independent experimental units for each hematological variable.

      Single-degree-of-freedom contrasts were used to measure treatment differences. These a priori contrasts were 0 ppm vs. 30 ppm; 30 ppm vs. 90 ppm and 30 ppm vs. 300 ppm sarafloxacin.

    6. Conclusion:

      Treatment related effects were biologically insignificant when sarafloxacin was administered in the drinking water for 15 days to turkeys at average levels of 33 to 342 ppm. There were no clinical signs of toxicity or illness among any of the groups. Body weight and feed consumption effects were generally positive, but not dose related.

      Water consumption effects were generally positive, but not dose-related. Water consumption was reduced during the period of medication at the highest level of sarafloxacin, but not during the post-medication period.

      Mean hematology values were within reported ranges for turkeys. Gross and microscopic tissue changes were mild and not dose-related.

      Therefore, the drug was demonstrated to be safe at 6X the maximum recommended dose and 10X the minimum recommended dose for 3X the recommended duration of treatment.

VI. HUMAN SAFETY

A. Toxicity Studies

  1. A Subchronic (3 month) Oral Toxicity Study in the Mouse with A-56620 via Dietary Admixture

    1. Report Number: 90-3575
    2. Study Completion: August 23, 1991 (Start: September 6, 1990)
    3. Investigators:
      J. E. Atkinson and Ira W. Daly
      Bio/dynamics Inc.
      East Milstone, NJ 08575
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) was used in this study.
    5. Species and Strain of Animal: CD-1 albino mice were used.
    6. Number of Animals per Group: Animals were allocated 50/sex/group.
    7. Levels and Duration of Dosing: CD-1 mice, approximately 6 weeks of age at initiation of the study, were given doses of 0, 20,000 or 50,000 ppm of sarafloxacin HCl. Animals were randomly assigned to treatment groups and scheduled for necropsy following treatment for 3 months.
    8. Route of Administration: The drug was given orally in the diet.
    9. Parameters Studied: Clinical observations, body weight, food consumption, and macroscopic pathology were determined.
    10. Toxicities Observed: Mortality occurred at 20,000 ppm and above.
    11. No Observable Effect Level (NOEL): This was not determined.
    12. Conclusions: Doses exceeded maximum tolerated dose and drug-related mortality was observed at all dose levels. Although this study was not pivotal in the usual sense in setting the NOEL, it was critical to the regulatory decision making process.
  2. Three-Month Toxicity (with One-Month Interim Kill) Study of Abbott-56620 Administered Orally to Rats

    1. Report Number: TA84-254
    2. Study Completion: July 15, 1985 (Start: November 6, 1984)
    3. Investigators:
      J. M. Creighton and M. C. Pratt
      Abbott Laboratories
      Abbott Park, Illinois 60064
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) in liquid suspension was used in this study.
    5. Species and Strain of Animal: Sprague-Dawley rats were used.
    6. Number of Animals per Group: Animals were allocated 20/sex/group and 5/sex/group.
    7. Levels and Duration of Dosing: Sprague-Dawley rats, approximately 6 weeks of age at initiation of the study, were given doses of 0, 20, 75, 275, or 1,000 mg/kg/day of sarafloxacin suspended in 0.2 percent hydroxypropylmethyl-cellulose (HPMC). Animals were randomly assigned to treatment groups and scheduled for necropsy following treatment for 1 month (10/sex/group) and 3months (10/sex/group). Animals assigned to a 1-month recovery phase (5/sex/group) were also scheduled for necropsy.
    8. Route of Administration: Drug was given by oral gavage.
    9. Parameters Studied: Clinical observations, body weight, food consumption, ophthalmoscopic examinations, urinalysis, hematology, clinical chemistry, organ weight, and gross and microscopic pathology were studied.
    10. Toxicities Observed: Mortality was observed at 1,000 mg/kg/day.
    11. No Observable Effect Level (NOEL): 275 mg/kg/day.
    12. Conclusions: Grossly enlarged ceca in males given 75 mg/kg/day and females given 275 mg/kg/day are believed to have resulted from the pharmacologic effects of large doses of the test substance. No treatment-related microscopic alterations were detected. This study was not used for setting the NOEL due to the small number of animals, but was still used in the regulatory decision making process.
  3. Thirteen-Week Capsule Toxicity Study with Abbott-56620 in Dogs

    1. Report Number: TB90-180
    2. Study Completion: April 26, 1991 (Start: July 20, 1990)
    3. Investigators:
      A. L. Kiorpes and R. H. Weltman
      Hazleton Wisconsin
      Madison, Wisconsin
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) gelatin capsules were used in this study.
    5. Species and Strain of Animal: Juvenile Beagle dogs were used in this study.
    6. Number of Animals per Group: 6/sex/group were used in the control and high-dosage groups; 4/sex/group were used in the low-and mid-dosage groups.
    7. Levels and Duration of Dosing: 0, 10, 50, or 200 mg/kg/day of sarafloxacin in gelatin capsules were given once daily for at least 13 weeks. During the first two weeks of the study, sarafloxacin, as the hydrochloride salt, was administered on an actual weight basis without regard to its free base concentration. This resulted in actual dosages which were approximately 80 percent of the intended free base dosages. For the remainder of the study, a correction factor was used in dose calculations to adjust for the free base concentration.
    8. Route of Administration: The drug was given orally as gelatin capsules.
    9. Parameters Studied: Clinical observations, body weight, food consumption, electroretinograms, ophthalmologic examinations, electrocardiograms, urinalysis, hematology, clinical chemistry, and gross and microscopic pathology were studied.
    10. Toxicities Observed: Transient localized dermal erythema and swelling, and ocular discharge were observed.
    11. No Observable Effect Level (NOEL): 8 mg/kg/day.
    12. Conclusions: Localized erythema and/or swelling was observed in dogs given 40 or 160 mg/kg/day, which is thought to occur in response to profound vasodilation caused by high doses of sarafloxacin or other quinolones. The phenomenon was transient and species specific. Increased incidence of ocular discharge was observed in females given 160 mg/kg/day. Lowered serum globulin values observed in treated dogs were not associated with morphologic changes representative of a systemic toxicologic response.
  4. Dietary Chronic Toxicity and Carcinogenicity Study of Abbott-56620 in the Albino Rat: Chronic Toxicity Phase

    1. Report Number: 83828 (TA88-435)
    2. Study Completion: October 16, 1990 (Start: December 16, 1988)
    3. Investigator: S. Y. Smith
      Bio Research Laboratories, Ltd.
      Montreal, Quebec, Canada
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620).
    5. Species and Strain of Animal: Sprague-Dawley derived CD rats were used.
    6. Number of Animals per Group: Animals were allocated 20/sex/dose.
    7. Levels and Duration of Dosing: Rats, approximately 6 weeks of age at initiation, were given 0, 1,000, 10,000, or 25,000 ppm sarafloxacin hydrochloride in the diet. The approximate mean average achieved was 54, 586, and 1,541 mg/kg/day for rats given diets containing 1,000, 10,000, or 25,000 ppm sarafloxacin hydrochloride, respectively. This was a 24-month study.
    8. Route of Administration: The drug was given orally, in the diet.
    9. Parameters Studied: Clinical observations, body weight, food consumption, ophthalmoscopic examinations, urinalysis, hematology, clinical chemistry, organ weight, and gross and microscopic pathology were studied.
    10. Toxicities Observed: Nephropathy was observed.
    11. No Observable Effect Level (NOEL): 1,000 ppm (approximately 54 mg/kg/day).
    12. Conclusions: Mild renal changes were observed in the 10,000 and 25,000 ppm dose groups, believed to be a result of drug insolubility. Lowered serum total protein and globulin values for treated rats were not associated with morphologic changes representative of a systemic toxicologic response. Cecal enlargement, a common result of antibiotic reduction of enteric microbes, was observed.
  5. Dietary Carcinogenicity Study of Abbott-56620 in the Albino Mouse

    1. Report Number: 84097
    2. Study Completion: November 22, 1991 (Start: July 27, 1989)
    3. Investigators:
      B. G. Procter, R. B. Salame, and J. W. Noveroske
      Bio Research Laboratories, Ltd.
      Montreal, Quebec, Canada
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) in the diet.
    5. Species and Strain of Animal: CD-1 albino mice were used.
    6. Number of Animals per Group: Animals were allocated 70/sex/group.
    7. Levels and Duration of Dosing: Mice were treated for approximately 78 weeks with diets containing 0, 1,000, 5,000, or 20,000 ppm sarafloxacin hydrochloride. Approximate mean average achieved intake of drug was 150, 830, and 3,300 mg/kg/day for mice given diets containing 1,000, 5,000, and 20,000 ppm, respectively.
    8. Route of Administration: Drug was administered orally in the diet.
    9. Parameters Studied: Clinical signs, body weights, food consumption, hematology, gross and microscopic pathology were studied.
    10. Toxicities Observed: Increased mortality, nephropathy were observed.
    11. No Observable Effect Level: 1,000 ppm (approximately 150 mg/kg/day).
    12. Conclusions: Administration of sarafloxacin hydrochloride to CD-1 mice for approximately 78 weeks had no carcinogenic effect. Nephropathic changes were observed in mice given 5,000 or 20,000 ppm. An increased incidence in mortality was observed in the 5,000 and 20,000 ppm groups. Dilated cecum, a pharmacologic response to the bactericidal action of large doses of this antibacterial, accompanied by mechanical rotation of the intestine may have contributed to the observed increase in mortality.
  6. A Dietary Chronic Toxicity and Carcinogenicity Study of Abbott-56620 In the Albino Rat: Carcinogenicity Phase

    1. Report Number: 83828 (TA88-435)
    2. Study Completion: December 26, 1991 (Start: December 16, 1988)
    3. Investigator:
      S. Y. Smith
      Bio Research Laboratories, Ltd.
      Montreal, Quebec, Canada
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) was used.
    5. Species and Strain of Animal: Sprague-Dawley CD rats were used.
    6. Number of Animals per Group: Animals were allocated 65/sex/group.
    7. Levels and Duration of Dosing: Rats were treated for 104 weeks with 0, 1,000, 10,000, or 25,000 ppm sarafloxacin hydrochloride. (Approximate mean average achieved intake of drug was 54, 586, and 1,541 mg/kg/day.)
    8. Route of Administration: Drug was given orally in the diet.
    9. Parameters Studied: Clinical signs, body weights, food consumption, ophthalmologic examinations, hematology, clinical chemistry, urinalysis, gross and microscopic pathology, and plasma drug level were studied.
    10. Toxicities Observed: Nephropathy was observed.
    11. No Observable Effect Level (NOEL): 1,000 ppm (approximately 54 mg/kg/day)
    12. Conclusions: Based on the results of this study, no evidence of carcinogenic activity was associated with the administration of sarafloxacin hydrochloride by dietary admixture to rats for a minimum of 104 weeks. The dosages of 10,000 and 25,000 ppm produced tubular nephropathy. Cecal enlargement, a common result of antibacterial reduction of enteric microbes, was observed.
  7. Evaluation of the Effects of Orally Administered Abbott-56620 on the Embryonic and Fetal Development of the Rat

    1. Report Number: TA84-279
    2. Study Completion: August 12, 1985 (Start: December 6, 1984)
    3. Investigator: S. B. Lehrer
      Abbott Laboratories
      Abbott Park, Illinois 60064
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) suspended in 0.2 percent HPMC.
    5. Species and Strain of Animal: Sperm-positive, female, Sprague-Dawley derived CD rats were used.
    6. Number of Animals per Group: 20 animals were allocated to each group.
    7. Levels and Duration of Dosing: Dosing with 0, 20, 75, 275, or 1,000 mg/kg/day of sarafloxacin started on the sixth day of gestation and ended after the fifteenth day of gestation.
    8. Route of Administration: Drug was administered orally by gavage.
    9. Parameters Studied: Potential maternal toxicity, embryo lethality, and teratogenicity were studied.
    10. Toxicities Observed: There were no drug-related, toxicologically meaningful changes in the parameters studied.
    11. No Observable Effect Level (NOEL): 1,000 mg/kg/day
    12. Conclusions: The mean numbers of viable fetuses/dam and implantation sites were comparable for all treatment groups when compared to controls. Fetal body weights and sex ratios were not significantly different from controls.
  8. Three-Generation Reproduction Study of Sarafloxacin (Abbott-56620) Administered Orally in Rats

    1. Report Number: WIL-57007 (TA88-116)
    2. Study Completion: May 7, 1991 (Start: October 5, 1987)
    3. Investigator:
      M. D. Nemec
      WIL Research Laboratories, Inc.
      Ashland, Ohio
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) suspended in 0.2 percent HPMC was used.
    5. Species and Strain of Animal: The drug was administered to three generations of rats, F0, F1, and F2. Each generation was permitted to produce up to two litters.
    6. Number of Animals per Group: Three groups, each consisting of 30 males and 30 females, were used.
    7. Levels and Duration of Dosing: 75, 275, and 1,000 mg/kg/day of sarafloxacin were administered for a minimum of 70 days prior to breeding.
    8. Route of Administration: The drug was administered orally by gavage.
    9. Parameters Studied: Clinical signs, body weight, food consumption, reproductive parameters, organ weight, and gross and microscopic pathology were studied.
    10. Toxicities Observed: There were no toxicologically meaningful changes in the parameters studied. However, lower liver weight was considered an indirect adverse effect related to treatment.
    11. No Observable Effect Level (NOEL): 75 mg/kg/day
    12. Conclusions: Reduced liver weight in rats treated with 275 or 1,000 mg/kg/day was dose-related, especially in females. The effect was considered an indirect adverse effect, caused by the drug (decreased cardiac output and, therefore, liver perfusion). No macroscopic or microscopic lesions of weanlings were attributed to parental treatment with sarafloxacin hydrochloride. Pups showed no adverse treatment-related effects. Litter size, pup survivability, pup weight, sex ratio, and general physical condition were not adversely affected.
  9. Evaluation of Abbott-56620, Lot 66-298-AL in the Rat Primary Hepatocyte Unscheduled DNA Synthesis Assay

    1. Report Number: 20991 (TA84-272)
    2. Study Completion: May 16, 1985 (Start: December 11, 1984)
    3. Investigator:
      M. A. Cifone
      Litton Bionetics, Inc.
      Kensington, Maryland
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) was used.
    5. Species and Strain of Animal: Fischer 344 rats were used.
    6. Number of Animals per Group: NA.
    7. Levels and Duration of Dosing: Hepatocytes were isolated from Fischer 344 rats by collagenase perfusion and, after establishment of primary hepatocyte cultures, the cultures were treated with concentrations of 0, 1, 2.5, 5.0, 10, 25, 50, 100, 250, and 500 ug/ml sarafloxacin for 18 hours. The positive control used in the study was 2-acetyl aminofluorene (0.01 ug/ml).
    8. Route of Administration: NA.
    9. Parameters Studied: During the treatment period the cultures were provided radiolabeled (tritium) thymidine, which, if incorporated into DNA, provides an indication of DNA synthesis activity. At the end of the treatment period the cultures were prepared for autoradiography.
    10. Toxicities Observed: NA.
    11. No Observable Effect Level (NOEL): NA.
    12. Conclusions: Significant increases in unscheduled DNA synthesis were induced at concentrations of 50, 100, and 250 ug/ml; the activity was dose-related. The in vitro UDS assay showed that sarafloxacin is clearly positive in UDS inducing activity. In addition, cytotoxicity defined as reduced cell survival, was apparent at a concentration of 250 ug/ml and was only minimally toxic at 100 ug/ml.
  10. Mutagenicity Test on Abbott-56620 in an In Vitro Cytogenetic Assay Measuring Chromosomal Aberration Frequencies in Chinese Hamster Ovary (CHO) Cells

    1. Report Number: HLA10161-0-437 (TG88-133)
    2. Study Completion: December 12, 1988 (Start: April 26, 1988)
    3. Investigator:
      H. Murli
      Hazleton Laboratories
      Kensington, Maryland
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) was used.
    5. Species and Strain of Animal: Chinese hamsters were used.
    6. Number of Animals per Group: NA.
    7. Levels and Duration of Dosing: Chinese hamster ovary dividing cell cultures were treated with sarafloxacin hydrochloride dissolved in dimethyl sulfoxide at concentrations of 0, 49.8, 99.6, 149, 199, 398, 598, and 797 µ g/ml for assays without metabolic activation and concentrations of 120, 299, 598, 897, 1,200, and 1,590 µ g/ml for assays with metabolic activation. The positive control agents used in the assays were mitomycin for nonactivation studies and cyclophosphamide in the metabolic activation assay. Treatment was conducted either with or without an exogenous metabolic activation system provided by a rat liver microsomal preparation.
    8. Route of Administration: NA.
    9. Parameters Studied: Chromosomal aberration frequency was studied.
    10. Toxicities Observed: NA.
    11. No Observable Effect Level (NOEL): NA.
    12. Conclusions: Under the conditions of these assays, sarafloxacin was shown to be reproducibly clastogenic with metabolic activation, but was considered negative for inducing chromosomal aberrations under nonactivated conditions. The positive clastogenic responses were statistically significant and reproducible in the presence of the metabolic supplement.
  11. Evaluation of Abbott-56620, Lot-66-298-AL, in the Forward Mutation Assay [at the HGPRT locus of Chinese Hamster Ovary (CHO) Cells]

    1. Report Number: LB22207 (T84-273)
    2. Study Completion: February 15, 1985 (Start: January 15, 1985)
    3. Investigator:
      R R. Young
      Litton Bionetics, Inc.
      Kensington, Maryland
    4. Substance and Dosage Form: Sarafloxacin HCl (A-56620) was used.
    5. Species and Strain of Animal: Chinese hamsters were used.
    6. Number of Animals per Group: NA.
    7. Levels and Duration of Dosing: Cultures of actively growing Chinese hamster ovary cells were treated with sarafloxacin hydrochloride for four hours either in the presence or absence of rat liver microsomal metabolic activation. The concentrations of sarafloxacin were 0, 50, 100, 200, 400, 600, 800, and

      1,000 ug/ml. The positive control used in the absence of metabolic activation was 5-bromo-2-deoxyuridine; the positive control used in the presence of metabolic activation was 3-methylcholanthrene.

    8. Route of Administration: NA.
    9. Parameters Studied: Mutation frequency was studied.
    10. Toxicities Observed: NA.
    11. No Observable Effect Level (NOEL): NA.
    12. Conclusions: In the presence of metabolic activation, a statistically significant increase in the number of mutations was seen at the highest dose tested, with smaller increases (2.4-, 3.6-, and 3.9-fold, relative to the concurrent negative control) at 3 of the remaining 5 lower dose levels. Based on these data, the compound is considered a suspect mutagen. The compound was negative without activation.

B. Safe Concentration of Total Residues

  1. No-Observed-Effect Level (NOEL):

    The safe concentration of total residue was determined from the lowest NOEL in the most sensitive species from the various toxicology studies conducted. A summary of the studies which can be used in determination of the Acceptable Daily Intake (ADI) follows:

    Table 6.1. Summary of ADI Toxicology Studies for Sarafloxacin HCl

    Study Type Species Doses mg/kg/day* NOEL mg/kg/day*

    90 day oral Dog 0, 10, 50, 200 10 Chronic Tox & Rat 0, 54, 586, 1541 54 Oncogenicity Oncogenicity Mouse 0, 150, 830, 3300 150 Segment II Rat 0, 20, 75, 275, 1000 75 Teratology Three-generation Rat 0, 75, 275, 1000 75 Reproduction

    *Doses based on sarafloxacin HCl bulk drug. The above values are uncorrected for 8 to 12% water content in the bulk drug.

    The 90-day oral dog study was selected as the most appropriate study with the lowest NOEL for determining the acceptable daily intake (ADI). The NOEL was 10 mg/kg bw/day sarafloxacin HCl bulk drug, which, when corrected for hydration, was calculated to be 8.75 mg/kg bw/day sarafloxacin HCl.

  2. Calculation of the Acceptable Daily Intake (ADI) and the Safe Concentration (SC) for Sarafloxacin HCl:

    1. Acceptable Daily Intake (ADI)

      ADI =     NOEL       =  8.75 mg/kg/day  =  0.00875 mg/kg/day  = 8.75 µg/kg/day
            safety factor         1000   
      
    2. Safe Concentration (SC)

      The calculation of the safe concentration is based on the General Principles for Evaluating the Safety of Compounds used in Food-Producing Animals (FDA, 1986):

      (SC) =  Acceptable Daily Intake (ADI) x Human Weight
                                 Food Factor
      
      Where:  Human Weight  = 60 kg
              Food Factor:    muscle = 300 g 
                              fat/skin = 50 g 
                              liver = 100g
                              
      SC (muscle) = 8.75 µg/kg/day x 60 kg = 1.75 µg/g = 1.75 ppm
                           300 g/day
                            
      SC (fat/skin) = 8.75 µg/kg/day x 60 kg = 10.5 µg/g = 10.5 ppm
                             50 g/day
                                
      SC (liver) = 8.75 µg/kg/day x 60 kg = 5.25 µg/g = 5.25 ppm
                          100 g/day
      

      Therefore, the safe concentration as total sarafloxacin HCl (ppm) in edible chicken or turkey tissue is:

      Tissue               Calculated Safe        
                           Concentration (ppm)
      Muscle                    1.75              
      Fat/Skin                 10.5              
      Liver                     5.25      
      
  3. Threshold Assessment

    Sarafloxacin HCl was positive in genotoxicity assays. Sarafloxacin was not carcinogenic in mouse and rat carcinogenicity bioassays.

C. Total Residue Depletion and Metabolism Studies

Investigators:
Jerry Johnson and Poonam Velagaleti
Battelle Institute
Columbus, OH 43201

Animals Used: 22 male and 22 female 3-week-old broilers, and 23 male and 23 female 8- to 12-week-old turkeys were used.

Drug was administered orally at 20 ppm and at 40 ppm to broilers or 30 ppm and at 60 ppm to turkeys for 5 consecutive days. (14)C sarafloxacin HCl was used for the study. The label was at a position not susceptible to metabolism.

Total residues (ppm) as sarafloxacin HCl equivalents in edible tissues (12 broiler chickens: 40 ppm) six hours after the last dose were:

Tissue          Calculated Safe           Mean Observed Total   
              Concentration* (ppm)        Residue (ppm)  ± SD  
Muscle               1.75                0.0537 ± 0.0125
Fat/Skin             10.5                0.0586 ± 0.0177
Liver                5.25                0.6980 ± 0.224

* As sarafloxacin HCl

Total residues (ppm) as sarafloxacin HCl equivalents in edible tissues (12 turkeys: 60 ppm) six hours after the last dose were:

Tissue          Calculated Safe            Mean Observed Total           
              Concentration* (ppm)         Residue (ppm) ± SD 
Muscle               1.75                 0.0459 ± 0.0495
Fat/Skin            10.5                  0.0665 ± 0.0585
Liver                5.25                 1.0574 ± 0.4953

* As sarafloxacin HCl

Metabolite characterization was carried out in the liver of broilers sacrificed at 6 hours following administration of 20 ppm (14)C-sarafloxacin for 5 days. Tissues were extracted and profiled using methods similar to those in the laboratory animals. The quantitation of the radioactivity is presented in Table 6.2.

Table 6.2. Quantitation of (14)C-Radioactive Components in Pooled Liver Tissue of Broilers Treated for 5 Days with 20 ppm (14)C-Sarafloxacin (% TRR: Percentage of Total (14)C-radioactive Residue; ppm: µg (14)C-Sarafloxacin Hydrochloride Equivalents/g Tissue)

                               Male       Male      Female    Female    
                               %TRR       ppm        %TRR      ppm
Sarafloxacin HCl               69.1       0.235      64.7      0.285
Total                           7.75      0.027      13.3      0.059
Sarafloxacin-conjugates
(acid labile)
Total Unidentified              8.48      0.029       9.03     0.040
Unextractable                  17.0       0.058      12.5      0.055
Total Recovery                102.3       0.348      99.6      0.438

The data indicate that parent sarafloxacin comprised an average of 67% of the total radioactive residue with 11% of the radioactivity a mixture of acid labile conjugates of sarafloxacin and 9% of the total radioactivity unidentified. Approximately 15% of the radioactivity was not extractable. Thus, sarafloxacin is only minimally biotransformed in the broiler chicken.

In the turkey, metabolite characterization was carried out in the liver of animals sacrificed at 6 hours following administration of (14)C-sarafloxacin for 5 days. Tissues were extracted and profiled using methods similar to those in the laboratory animals. The quantitation of the radioactivity is presented in Table 6.3.

Table 6.3. Quantitation of (14)C-Radioactive Components in Pooled Liver Tissue of Turkeys Treated for 5 Days with 30 ppm (14)C-Sarafloxacin. (% TRR: Percentage of Total Radioactive Residue, ppm: µg (14)C-Sarafloxacin Hydrochloride Equivalent/g Tissue)

                         Male          Male        Female     Female       
Components              % TRR          ppm         % TRR       ppm
Sarafloxacin             20.4          0.07         20.9       0.13
Sarafloxacin              6.8          0.02          6.1       0.04
Sulfamic Acid       
Sarafloxacin             20.4          0.06         25.1       0.16
Glucuronide        
Sarafloxacin             29.8          0.09         16.0       0.10       
Sulfamic                                                                   
Glucuronide
Others (4 Minor           6.2          0.02         15.2       0.09        
Metabolites)
Unextractable            18.2          0.06         16.0       0.10
Total Recovery          101.8          0.32         99.3       0.62 

The data indicate that very little primary metabolism occurred in the turkey; however, sarafloxacin was extensively conjugated in this species. Approximately 21% of radioactivity present in liver was parent sarafloxacin. Sarafloxacin glucuronide, sarafloxacin sulfamic glucuronide and sarafloxacin sulfamic acid comprised 25, 16 and 6%, respectively, of the total radioactive residue in the liver. Four (4) minor metabolites accounted for 10% of the radioactivity in the liver; these were converted to sarafloxacin by acid or base hydrolysis and are considered conjugates. A total of 17% of the radioactivity in the liver was unextractable.

D. Comparative Metabolism

The metabolism of sarafloxacin was studied in the rat, dog and mouse. Methods for analysis in these studies were similar to those used for the target animal species. Following administration of a 10 mg/kg single oral dose of (14)C-sarafloxacin to rats, radioactivity was excreted in the urine and feces, where it comprised 37 and 52% of the total administered drug, respectively. Of this, the majority was unchanged parent: 99-100% of eliminated activity in the urine and feces. Overall, parent drug in the excreta accounted for 86% of the dose. Small amounts of N-acetyl sarafloxacin and a 3'-oxo compound were also identified in the urine and feces. In the dog, a 10 mg/kg dose of (14)C-sarafloxacin was eliminated primarily in the urine (60%) with 31% of the administered radioactivity eliminated in the feces. Of this, 91 and 83% of the radioactivity in the urine and feces, respectively, was parent drug. Thus a total of 79% of the administered dose was eliminated as parent drug in the dog. No other metabolites were identified. In a separate study, 13 and 5% of a 10 mg/kg (14)C-sarafloxacin dose administered intravenously or intraduodenally to dogs, respectively, were collected in the bile within 6 hours of administration. Approximately half of the radioactivity in the bile was unchanged parent, the other half sarafloxacin glucuronide.

Mice were administered a single oral dose of 100 mg/kg (14)C-sarafloxacin. Unchanged parent in the urine comprised 11% of the administered dose, with 5% of the dose in the form of sarafloxacin glucuronide. N-acetyl sarafloxacin was also present as a minor metabolite in the mouse. Sarafloxacin was primarily excreted in the feces, where it comprised 71% of the administered radioactivity. The metabolism of a 100 mg/kg dose in mice was similar to that of a 10 mg/kg dose.

It can be seen that very little primary metabolism takes place in either target animals or laboratory animals treated with sarafloxacin. The predominant transformation of sarafloxacin is conjugation in all species tested. There are no major metabolites in the broiler. There are two major metabolites in the turkey: sarafloxacin glucuronide and sarafloxacin sulfamic glucuronide. The glucuronide has been identified in two laboratory animals, mice and dogs. Although the sarafloxacin sulfamic glucuronide has not been identified in laboratory animals, it is present in low amounts (<0.1 ppm) in only the turkey and, as a conjugate, is considered to have toxicity similar to that of the parent.

E. Withdrawal Time

The total residue data showed that the mean concentrations of total sarafloxacin residues at 6 hours after withdrawal ("zero-day withdrawal") were well below half of the permitted safe concentration in the edible tissues of growing broiler chickens and turkeys treated with 40 and 60 ppm sarafloxacin, respectively. Therefore, a withdrawal period will not be required for this use of the drug in poultry.

F. Regulatory Method

An official regulatory method is not required because the residue and toxicology data support a zero-day withdrawal.

G. Target Tissue and Tolerance

Although an official tolerance is not established, the following information is provided for reference purposes. At zero withdrawal the total residue of sarafloxacin HCl in the target tissue (liver) is less than half the safe concentration (5.25 ppm). The marker compound, parent sarafloxacin HCl, represents 20 to 80% of the total residue in liver of turkeys and 60 to 80% of the total residue in liver of chickens, depending upon the extraction procedure.

H. User Safety Concerns

Sarafloxacin HCl was consistently nontoxic and nonlethal at maximally achievable concentrations following single oral or dermal exposure. Dermal exposure of 2 g/kg was shown to be noncorrosive and nonirritating to rabbit skin. Ocular exposure of 0.1 g in the rabbit eye was non-corrosive and non-irritating. Conjunctival redness completely reversed within 48 hours. Sarafloxacin HCl is not a dermal sensitization agent. In the guinea pig maximization test, the drug produced no evidence of delayed hypersensitivity.

User safety concerns associated with hypersensitivity to quinolones or photosensitization due to accidental inhalation or direct contact have been satisfactorily addressed by establishing label warnings. In addition, a toll free telephone number will be added to the label to inform users of where they can obtain additional information concerning user safety relative to the MSDS and to report adverse effects.

 

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 21 CFR Part 514 of the implementing regulations. The data demonstrate that sarafloxacin hydrochloride water soluble powder (SaraFlox® WSP), a fluoroquinolone antibiotic, when administered ad libitum in the drinking water at a concentration of 20 to 40 ppm for growing broilers, or 30 to 50 ppm for growing turkeys for five consecutive days, is safe and effective for the treatment of colibacillosis caused by Escherichia coli.

Based on a battery of toxicology tests, the safe concentrations for total residues of sarafloxacin HCl are 1.75 ppm in muscle, 5.25 ppm in liver, and 10.5 ppm in fat/skin.

The total residue data showed that the mean concentrations of total sarafloxacin residues at 6 hours after withdrawal ("zero-day withdrawal") were well below half of the permitted safe concentration in the edible tissues of growing broiler chickens and turkeys treated with 40 and 60 ppm sarafloxacin, respectively. Therefore, a withdrawal period will not be required for this use of the drug in poultry,.

An official regulatory method is not required because the residue and toxicology data support a zero-day withdrawal.

Labeling restricts this drug to use by or on order of a licensed veterinarian. This decision was based on the following factors: (a) the product contains a new antimicrobial entity intended only for therapeutic purposes, (b) adequate directions cannot be written to enable lay persons to appropriately diagnose and subsequently use this product to treat colibacillosis, and (c) the frequency of violative tissue residues and rate of emergence of sarafloxacin-resistant organisms will be reduced by the involvement of veterinarians in product use.

Public health concerns associated with potential increases in antimicrobial resistance have been satisfactorily addressed by establishing conditions of use intended to minimize inappropriate use of this product, and excretion of sarafloxacin and sarafloxacin-resistant zoonotic pathogens into the environment. The sponsor has agreed to participate in a national bacterial resistance surveillance program, and has provided preapproval baseline susceptibility information on potential human pathogens known to contaminate animal carcasses.

The agency has carefully considered the potential environmental effects of this action and has concluded that the action will not have a significant impact on the human environment and that and environmental impact statement is not required. The agency's finding of no significant impact (FONSI) and the evidence supporting that finding are contained in an environmental assessment, which may be seen in the Docket Management Branch (HFV-305), Park Building (Room 1-23), 12420 Parklawn Dr., Rockville, Maryland 20855.

Section 512(c)(2)(F)(i) of the act provides a five-year exclusivity period for a drug, no active ingredient (including any ester or salt of the active ingredient) of which has been approved in any other application under subsection (b)(1). This NADA qualifies for such an exclusivity period.

Sarafloxacin hydrochloride is under patent number U.S. 4,730,000 expiring March 18, 2005.

 

VIII. APPROVED PRODUCT LABELING

A copy of the draft facsimile labeling is attached to this document.

  1. SaraFlox® Water Soluble Powder Pail Label Copy
  2. SaraFlox® Water Soluble Powder Pouch Front Copy
  3. SaraFlox® Water Soluble Powder Pouch Back Copy
  4. Package Insert Page 1
  5. Package Insert Page 2

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