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Chapter 8. Synthetic Antibacterials
【Outline of synthetic antibacterials】
Antibacterial substance is one of feed additives to be used in order to promote beneficial use of
nutrient components in feed. “Antibacterial substance” is a collective term of substances having antibacterial
activity, which includes synthetic antibacterials (synthesized antibacterials specified as a feed additive are
referred to as “synthetic antibacterial agents”), antibiotics as well as, fungicides such as propionic acid
specified as antibacterial substances in Feed Safety Act. However, in general, this term is used as that
including synthetic antibacterial agents and antibiotics permitted to use in order to increase the productivity
by improving the growth and feed conversion and by disinfesting Coccidia and endoparasites, and those
synthesized chemically are defined as synthetic antibacterials being discriminated from antibiotics which are
made from microorganisms.
The standards based on Feed Safety Act specified that feeds never contain antibacterial substances
other than those set down as feed additives.
Easy use of antibacterial substances can cause public health problems such as the residue in animal
products and emergence of drug-resistant strains of bacteria even if they have been set down as feed
additives; therefore, there are many items to keep in mind at the time of use to be strictly controlled in
respective steps, manufacture, distribution and usage. The basic items which persons handling feed additives
must know are as follows.
I Synthetic antibacterial agents and antibiotics available for respective animal species, types of feed, and
additive amounts were specified individually; should not be used in other feed than specified.
II Synthetic antibacterial agents and antibiotics should not be used in feeds for cultured fish.
III Those capable and incapable of combination use of 2 types or more are specified, respectively.
IV Antibiotics are designated to the specific feed additive, which cannot be sold except for those with a
special label describing that the article has approved by FAMIC or has manufactured in a facility
licensed by the Minister of Agriculture, Forestry and Fisheries.
V Synthetic antibacterial agents and antibiotics cannot be used for animals other than indicated by
description on the package.
VI Feeds containing synthetic antibacterial agent(s) and/or antibiotic(s) should not be supplied to milking
cows, laying hens or quails, or domestic animals or chickens to be killed for edible use within 7 days.
Target animal types, usage levels and the like for each synthetic antibacterial agent were specified
by Ministerial Ordinance Concerning Compositional Standards, and the specifications are sequentially
canceled or added. As of November, 2009, a total of 6 articles (7 components) including amprolium
ethopabate, amprolium ethopabate sulfaquinoxaline, morantel citrate, decoquinate, nicarbazin and
halofujinon calcium polystyrene-sulfonate have been specified as the synthetic antibacterial agents.
As for canceled articles, some of them are still in distribution as animal drugs, and the analysis
methods have been available in the Feed Analysis Standards because of possible use as feed additives.
Furthermore, this section also specifies the analytical methods of residual synthetic antibacterials in
animal feeds for articles possibly used abroad for domestic animals or cultured fish.
Standard preparations of synthetic antibacterial agents specified as feed additive are commercially
available at Japan Scientific Feeds Association (TEL: 03-3297-5631, FAX: 03-3297-5633)
《Summary Table of analytical methods of synthetic antibacterials》
Analytical methods of synthetic antibacterials which currently listed in Feed Analysis Standards
(FAS) are summarized in Tables 8-1 to 8-3.
Table 8-1 Summary Table of analytical methods of synthetic antibacterials
Micro-
LC method determination
Premix 1.1.1(1) 1.1.2(1) Absorbance
Formula feed 1.1.1(2) 1.1.2(2) Absorbance 1.2.1
Premix 2.2.1(1)
Formula feed 2.2.1(2) 2.2.1
Premix 7.1.1(1) 7.2.1
Formula feed 7.1.1(2) 7.2.1
Premix 10.1.1(1) 10.2.1
Formula feed 10.1.1(2) 10.2.1
Premix 11.1.1(1) 11.1.2(1) Fluorometry 11.2.1(1)
Formula feed 11.1.1(2) 11.1.2(2) Fluorometry 11.2.1(2)
Premix 12.1.1(1) 12.1.2(1) Absorbance
Formula feed 12.1.1(2) 12.1.2(2) Absorbance 12.2.1
Halofujinon calcium Premix 13.1.1(1)
 polystyrene-sulfonate Formula feed 13.1.1(2) 13.2.1 Control assay (Absorbance)
Quantification (Absorbance)
Other mrthod
24/7/1976
Absorbance/fluorometry
15/10/1985
Nicarbazin
Analytical method listed in FAS Note
Amprolium
Scope of
application
Morantel citrate
Sulfaquinoxaline
Decoquinate
24/7/1976Ethopabate
Quantification to labeled amount
25/12/1987
24/7/1976
24/7/1976
24/7/1976
Name of synthetic
antibacterial agent
Date of designation
Note: Figures indicate the listing no. (Feed Analysis Standards Chapter 8, Section 1)
Table 8-2 Summary Table of analytical methods of synthetic antibacterials cancelled from
specification as a feed additive
Micro-
LC method determination
Premix 3.1 Absorbance
Formula feed 3.1 Absorbance
Premix 5.1.1(1) 5.2.1
Formula feed 5.1.1(2) 5.2.1
Premix 6.1 Absorbance
Formula feed 6.1 Absorbance Quantification (LC method)
Premix 8.1
Formula feed 8.1
Premix 9.1
Formula feed 9.1
Furazolidone Note 2 Formula feed 14.2
27/7/1981
20/3/2002
Dinitolmide
Clopidol
Analytical method listed in FASNote 1
Quantification to added amount
Name of synthetic
antibacterial agent
Date of cancel
27/7/1981
Absorbance/fluorometry
Other method
7/3/2001
Scope of
application
27/7/1981
Lobenidine hydrochloride
Oraquindox
Carbadox
Note 1: Figures indicate the listing no. (Feed Analysis Standards Chapter 8, Section 1).
Note 2: Cancelled from specification as a feed additive at the time of revision of Feed Safety Act, in1976,
when feed additives became designation basis by Minister of Agriculture, Forestry and Fisheries.
See Paragraph 14, Section 1 in this Chapter.
Table 8-3 Summary Table of analytical methods of other synthetic antibacterials
LC method LC-MS/MS method
Oxolinic acid Fish meal and formula feed 4.1
Flumequine Fish meal and formula feed 15.1
Fish meal and formula feed 16.1
Fish oil 16.2
Leucomalachite Fish meal and formula feed 17.1
green Fish oil 17.2
Simultaneous determination
(Paragraph 2, Section 2)
Simultaneous determination
(Paragraph 3, Section 2)
Listing no. in FAS
Reference
Malachite green
Scope of application
Simultaneous determination
(Paragraph 2, Section 2)
Name of synthetic
antibacterial agent
Simultaneous determination
(Paragraph 2, Section 2)
Simultaneous determination
(Paragraph 3, Section 2)

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Synthetic antibacterial agents

  • 1. Chapter 8. Synthetic Antibacterials 【Outline of synthetic antibacterials】 Antibacterial substance is one of feed additives to be used in order to promote beneficial use of nutrient components in feed. “Antibacterial substance” is a collective term of substances having antibacterial activity, which includes synthetic antibacterials (synthesized antibacterials specified as a feed additive are referred to as “synthetic antibacterial agents”), antibiotics as well as, fungicides such as propionic acid specified as antibacterial substances in Feed Safety Act. However, in general, this term is used as that including synthetic antibacterial agents and antibiotics permitted to use in order to increase the productivity by improving the growth and feed conversion and by disinfesting Coccidia and endoparasites, and those synthesized chemically are defined as synthetic antibacterials being discriminated from antibiotics which are made from microorganisms. The standards based on Feed Safety Act specified that feeds never contain antibacterial substances other than those set down as feed additives. Easy use of antibacterial substances can cause public health problems such as the residue in animal products and emergence of drug-resistant strains of bacteria even if they have been set down as feed additives; therefore, there are many items to keep in mind at the time of use to be strictly controlled in respective steps, manufacture, distribution and usage. The basic items which persons handling feed additives must know are as follows. I Synthetic antibacterial agents and antibiotics available for respective animal species, types of feed, and additive amounts were specified individually; should not be used in other feed than specified. II Synthetic antibacterial agents and antibiotics should not be used in feeds for cultured fish. III Those capable and incapable of combination use of 2 types or more are specified, respectively. IV Antibiotics are designated to the specific feed additive, which cannot be sold except for those with a special label describing that the article has approved by FAMIC or has manufactured in a facility licensed by the Minister of Agriculture, Forestry and Fisheries. V Synthetic antibacterial agents and antibiotics cannot be used for animals other than indicated by description on the package. VI Feeds containing synthetic antibacterial agent(s) and/or antibiotic(s) should not be supplied to milking cows, laying hens or quails, or domestic animals or chickens to be killed for edible use within 7 days. Target animal types, usage levels and the like for each synthetic antibacterial agent were specified by Ministerial Ordinance Concerning Compositional Standards, and the specifications are sequentially canceled or added. As of November, 2009, a total of 6 articles (7 components) including amprolium ethopabate, amprolium ethopabate sulfaquinoxaline, morantel citrate, decoquinate, nicarbazin and halofujinon calcium polystyrene-sulfonate have been specified as the synthetic antibacterial agents. As for canceled articles, some of them are still in distribution as animal drugs, and the analysis methods have been available in the Feed Analysis Standards because of possible use as feed additives. Furthermore, this section also specifies the analytical methods of residual synthetic antibacterials in
  • 2. animal feeds for articles possibly used abroad for domestic animals or cultured fish. Standard preparations of synthetic antibacterial agents specified as feed additive are commercially available at Japan Scientific Feeds Association (TEL: 03-3297-5631, FAX: 03-3297-5633) 《Summary Table of analytical methods of synthetic antibacterials》 Analytical methods of synthetic antibacterials which currently listed in Feed Analysis Standards (FAS) are summarized in Tables 8-1 to 8-3. Table 8-1 Summary Table of analytical methods of synthetic antibacterials Micro- LC method determination Premix 1.1.1(1) 1.1.2(1) Absorbance Formula feed 1.1.1(2) 1.1.2(2) Absorbance 1.2.1 Premix 2.2.1(1) Formula feed 2.2.1(2) 2.2.1 Premix 7.1.1(1) 7.2.1 Formula feed 7.1.1(2) 7.2.1 Premix 10.1.1(1) 10.2.1 Formula feed 10.1.1(2) 10.2.1 Premix 11.1.1(1) 11.1.2(1) Fluorometry 11.2.1(1) Formula feed 11.1.1(2) 11.1.2(2) Fluorometry 11.2.1(2) Premix 12.1.1(1) 12.1.2(1) Absorbance Formula feed 12.1.1(2) 12.1.2(2) Absorbance 12.2.1 Halofujinon calcium Premix 13.1.1(1)  polystyrene-sulfonate Formula feed 13.1.1(2) 13.2.1 Control assay (Absorbance) Quantification (Absorbance) Other mrthod 24/7/1976 Absorbance/fluorometry 15/10/1985 Nicarbazin Analytical method listed in FAS Note Amprolium Scope of application Morantel citrate Sulfaquinoxaline Decoquinate 24/7/1976Ethopabate Quantification to labeled amount 25/12/1987 24/7/1976 24/7/1976 24/7/1976 Name of synthetic antibacterial agent Date of designation Note: Figures indicate the listing no. (Feed Analysis Standards Chapter 8, Section 1) Table 8-2 Summary Table of analytical methods of synthetic antibacterials cancelled from specification as a feed additive Micro- LC method determination Premix 3.1 Absorbance Formula feed 3.1 Absorbance Premix 5.1.1(1) 5.2.1 Formula feed 5.1.1(2) 5.2.1 Premix 6.1 Absorbance Formula feed 6.1 Absorbance Quantification (LC method) Premix 8.1 Formula feed 8.1 Premix 9.1 Formula feed 9.1 Furazolidone Note 2 Formula feed 14.2 27/7/1981 20/3/2002 Dinitolmide Clopidol Analytical method listed in FASNote 1 Quantification to added amount Name of synthetic antibacterial agent Date of cancel 27/7/1981 Absorbance/fluorometry Other method 7/3/2001 Scope of application 27/7/1981 Lobenidine hydrochloride Oraquindox Carbadox Note 1: Figures indicate the listing no. (Feed Analysis Standards Chapter 8, Section 1). Note 2: Cancelled from specification as a feed additive at the time of revision of Feed Safety Act, in1976, when feed additives became designation basis by Minister of Agriculture, Forestry and Fisheries.
  • 3. See Paragraph 14, Section 1 in this Chapter. Table 8-3 Summary Table of analytical methods of other synthetic antibacterials LC method LC-MS/MS method Oxolinic acid Fish meal and formula feed 4.1 Flumequine Fish meal and formula feed 15.1 Fish meal and formula feed 16.1 Fish oil 16.2 Leucomalachite Fish meal and formula feed 17.1 green Fish oil 17.2 Simultaneous determination (Paragraph 2, Section 2) Simultaneous determination (Paragraph 3, Section 2) Listing no. in FAS Reference Malachite green Scope of application Simultaneous determination (Paragraph 2, Section 2) Name of synthetic antibacterial agent Simultaneous determination (Paragraph 2, Section 2) Simultaneous determination (Paragraph 3, Section 2)