Florida Regulations 5C-6.0011: Laboratory Testing Procedures and Interpretation
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(1) The following methods of testing shall be considered as screening tests:
(a) Milk Ring Test (BRT). This test is conducted by a state-federal laboratory on composite samples of milk collected at dairy farms. A follow-up individual serological test shall be conducted on all cattle represented in a composite sample which reacts to the test. The milk ring test may be used in conjunction with a negative herd blood test to establish a Certified Brucellosis-Free Herd.
(b) Buffered Acidified Plate Antigen (BAPA) presumptive test. This test will be used by a state-federal laboratory to classify cattle and bison negative on surveillance samples collected at slaughter or at livestock markets, on routine samples collected on farms, and on tests of suspicious and infected herds. Positive samples will be given supplemental testing to aid in classification.
(c) Standard Card Test. This test will be used by a state-federal laboratory to classify cattle and bison negative on surveillance samples collected at slaughter or at livestock markets, on routine samples collected on farms, and on tests of suspicious and infected herds. Positive samples will be given supplemental testing to aid in classification.
(d) Particle Concentration Fluorescence Immunoassay (PCFIA). All samples with results within the suspect range will be given supplemental testing to aid in classification.
(2) The following methods of testing when conducted in a state-federal laboratory shall be considered as official tests for the classification of cattle and bison with regard to brucellosis status:
(a) Standard tube agglutination test. This test may classify cattle and bison as negative, suspect, or reactor.
(b) Standard plate agglutination test. This test may classify cattle and bison as negative, suspect, or reactor.
(c) Standard card test. The standard card test is to be used as an official test to classify cattle and bison as reactors only when conditions and time are such that no other test is available, or upon request of the owner because of time or situation constraints, or in livestock markets where the standard card test has been designated as the official test. All samples tested at other than a state-federal laboratory shall be promptly submitted and confirmed at a state-federal laboratory.
(d) Rivanol test. This test may classify cattle and bison as negative, suspect, or reactor.
(e) Complement-fixation test. This test may classify cattle and bison as negative, suspect, or reactor.
(f) Semen plasma test. This test may classify bulls used for artificial insemination as negative or reactor when used in conjunction with an official serological test.
(g) Particle Concentration Fluorescence Immunoassay (PCFIA). This test may classify cattle and bison as negative, suspect, or reactor. All samples with results within the suspect range will be given supplemental testing to aid in classification.
(3) The following diagnostic procedures shall be considered as supplemental tests and, when requested by the designated epidemiologist, shall be used to aid in determining the presence or absence of blood components associated with field strains of Brucella abortus, Brucella abortus strain 19, nonspecific agglutinins, etc.:
(a) Mercaptoethanol (ME)
(b) Coombs
(c) Heat inactivation (HI)
(d) Fluorescent Antibody (FA)
(e) Enzyme Labeled Antibody (ELA)
(f) Biological culture
(g) Acid Plate Antigen (APA)
(h) Concentration Immunoassay Technology (CITE)
(4) Standard tube agglutination test and standard plate agglutination test.
(a) Non-vaccinates. Blood titer showing complete reaction in the 1:100 dilution or higher shall be classed as a reactor. Titer of not more than incomplete in the 1:100 dilution shall be classed as a suspect. Titer less than incomplete in the 1:50 dilution shall be considered negative.
(b) Official vaccinates. Blood titer showing complete reaction in the 1:200 dilution or higher shall be classed as a reactor. Titer showing any reaction in the 1:100 dilution or incomplete in the 1:200 dilution shall be classed as a suspect. Titer less than incomplete in the 1:100 dilution shall be considered negative.
(5) Standard card test. Blood showing a positive reaction to the standard card test will be classed as a reactor when only the standard card test has been conducted. Cattle and bison positive to the standard card test which are serologically negative to all other official tests which are conducted are classified as suspects.
(6) Rivanol test.
(a) The rivanol test is an official test when conducted in a state-federal laboratory. Complete agglutination at dilutions of 1:25 or more indicates a reactor in nonvaccinated animals and in vaccinated animals when the complement-fixation test is not conducted. Less than complete agglutination at the 1:25 dilution shall be considered negative.
(b) Officially vaccinated animals with agglutination complete in the 1:50 dilution or less may be classified as suspects when those interpretations are supported by complement-fixation test results of suspect or negative using the presently accepted complement-fixation test classification scheme for vaccinated animals.
(c) Incomplete or complete agglutination in the 1:100 dilution or in higher dilutions indicates a reactor in vaccinated animals.
(d) Within five months postvaccination of adult animals, a less than complete agglutination at the 1:50 dilution is considered negative.
(7) Complement-fixation test.
(a) Official vaccinates, including adult vaccinated animals beginning two months postvaccination. Animals with titers of 25 percent fixation (1 plus) in a dilution of 1:40 or higher will be classified as reactors. Animals having less than 25 percent fixation (1 plus) in 1:40 dilution, and 50 percent (2 plus) or more in a dilution of 1:10 are classified as suspects. A reaction of less than 50 percent fixation (2 plus) in the 1:10 dilution is considered negative.
(b) Other cattle and bison. A 50 percent fixation (2 plus) in a dilution of 1:20 or higher will be classed as a reactor; fixation of less than 50 percent (2 plus) in a dilution of 1:20 and 50 percent fixation (2 plus) or more in a dilution of 1:10 will be classed as a suspect; less than 50 percent fixation (2 plus) in a dilution of 1:10 will be classed as negative.
(8) Particle Concentration Fluorescence Immunoassay (PCFIA). This test may be used as a presumptive or diagnostic test with the following diagnostic criteria:
(a) Negative – values greater than 0.70.
(b) Suspect – values less than or equal to 0.70 and greater than 0.25.
(c) Reactor – values less than or equal to 0.25.
(d) Values may be interpreted differently if done by a veterinarian, qualified by education, training, and experience and certified by the United States Department of Agriculture with the concurrence of the state and termed designated epidemiologist.
(9) Semen plasma test. This test in bulls used for artificial insemination must be used in conjunction with an official serological test, and classification will be based on the maximum titer of either test.
(10) Any animal found to be infected through isolation of field strain Brucella abortus must be classed as a reactor.
(11) Evaluation of titer responses for all cattle and bison shall be the responsibility of a designated epidemiologist, who takes into consideration animal and herd history and other epidemiologic considerations.
(12) Interpretation of market tests. Blood samples collected at slaughter are considered negative if no evidence of agglutination is disclosed on the buffered acidified plate antigen test. Samples showing agglutination are classified negative if the confirmatory standard card test is negative.
Rulemaking Authority Florida Statutes § 585.002(4), 585.08(2) FS. Law Implemented 585.08(1), (3), 585.11(1), (2), 585.155(1), (2), (3), (4) FS. History-New 6-1-92, Amended 7-5-95.
Terms Used In Florida Regulations 5C-6.0011
- Evidence: Information presented in testimony or in documents that is used to persuade the fact finder (judge or jury) to decide the case for one side or the other.
(b) Buffered Acidified Plate Antigen (BAPA) presumptive test. This test will be used by a state-federal laboratory to classify cattle and bison negative on surveillance samples collected at slaughter or at livestock markets, on routine samples collected on farms, and on tests of suspicious and infected herds. Positive samples will be given supplemental testing to aid in classification.
(c) Standard Card Test. This test will be used by a state-federal laboratory to classify cattle and bison negative on surveillance samples collected at slaughter or at livestock markets, on routine samples collected on farms, and on tests of suspicious and infected herds. Positive samples will be given supplemental testing to aid in classification.
(d) Particle Concentration Fluorescence Immunoassay (PCFIA). All samples with results within the suspect range will be given supplemental testing to aid in classification.
(2) The following methods of testing when conducted in a state-federal laboratory shall be considered as official tests for the classification of cattle and bison with regard to brucellosis status:
(a) Standard tube agglutination test. This test may classify cattle and bison as negative, suspect, or reactor.
(b) Standard plate agglutination test. This test may classify cattle and bison as negative, suspect, or reactor.
(c) Standard card test. The standard card test is to be used as an official test to classify cattle and bison as reactors only when conditions and time are such that no other test is available, or upon request of the owner because of time or situation constraints, or in livestock markets where the standard card test has been designated as the official test. All samples tested at other than a state-federal laboratory shall be promptly submitted and confirmed at a state-federal laboratory.
(d) Rivanol test. This test may classify cattle and bison as negative, suspect, or reactor.
(e) Complement-fixation test. This test may classify cattle and bison as negative, suspect, or reactor.
(f) Semen plasma test. This test may classify bulls used for artificial insemination as negative or reactor when used in conjunction with an official serological test.
(g) Particle Concentration Fluorescence Immunoassay (PCFIA). This test may classify cattle and bison as negative, suspect, or reactor. All samples with results within the suspect range will be given supplemental testing to aid in classification.
(3) The following diagnostic procedures shall be considered as supplemental tests and, when requested by the designated epidemiologist, shall be used to aid in determining the presence or absence of blood components associated with field strains of Brucella abortus, Brucella abortus strain 19, nonspecific agglutinins, etc.:
(a) Mercaptoethanol (ME)
(b) Coombs
(c) Heat inactivation (HI)
(d) Fluorescent Antibody (FA)
(e) Enzyme Labeled Antibody (ELA)
(f) Biological culture
(g) Acid Plate Antigen (APA)
(h) Concentration Immunoassay Technology (CITE)
(4) Standard tube agglutination test and standard plate agglutination test.
(a) Non-vaccinates. Blood titer showing complete reaction in the 1:100 dilution or higher shall be classed as a reactor. Titer of not more than incomplete in the 1:100 dilution shall be classed as a suspect. Titer less than incomplete in the 1:50 dilution shall be considered negative.
(b) Official vaccinates. Blood titer showing complete reaction in the 1:200 dilution or higher shall be classed as a reactor. Titer showing any reaction in the 1:100 dilution or incomplete in the 1:200 dilution shall be classed as a suspect. Titer less than incomplete in the 1:100 dilution shall be considered negative.
(5) Standard card test. Blood showing a positive reaction to the standard card test will be classed as a reactor when only the standard card test has been conducted. Cattle and bison positive to the standard card test which are serologically negative to all other official tests which are conducted are classified as suspects.
(6) Rivanol test.
(a) The rivanol test is an official test when conducted in a state-federal laboratory. Complete agglutination at dilutions of 1:25 or more indicates a reactor in nonvaccinated animals and in vaccinated animals when the complement-fixation test is not conducted. Less than complete agglutination at the 1:25 dilution shall be considered negative.
(b) Officially vaccinated animals with agglutination complete in the 1:50 dilution or less may be classified as suspects when those interpretations are supported by complement-fixation test results of suspect or negative using the presently accepted complement-fixation test classification scheme for vaccinated animals.
(c) Incomplete or complete agglutination in the 1:100 dilution or in higher dilutions indicates a reactor in vaccinated animals.
(d) Within five months postvaccination of adult animals, a less than complete agglutination at the 1:50 dilution is considered negative.
(7) Complement-fixation test.
(a) Official vaccinates, including adult vaccinated animals beginning two months postvaccination. Animals with titers of 25 percent fixation (1 plus) in a dilution of 1:40 or higher will be classified as reactors. Animals having less than 25 percent fixation (1 plus) in 1:40 dilution, and 50 percent (2 plus) or more in a dilution of 1:10 are classified as suspects. A reaction of less than 50 percent fixation (2 plus) in the 1:10 dilution is considered negative.
(b) Other cattle and bison. A 50 percent fixation (2 plus) in a dilution of 1:20 or higher will be classed as a reactor; fixation of less than 50 percent (2 plus) in a dilution of 1:20 and 50 percent fixation (2 plus) or more in a dilution of 1:10 will be classed as a suspect; less than 50 percent fixation (2 plus) in a dilution of 1:10 will be classed as negative.
(8) Particle Concentration Fluorescence Immunoassay (PCFIA). This test may be used as a presumptive or diagnostic test with the following diagnostic criteria:
(a) Negative – values greater than 0.70.
(b) Suspect – values less than or equal to 0.70 and greater than 0.25.
(c) Reactor – values less than or equal to 0.25.
(d) Values may be interpreted differently if done by a veterinarian, qualified by education, training, and experience and certified by the United States Department of Agriculture with the concurrence of the state and termed designated epidemiologist.
(9) Semen plasma test. This test in bulls used for artificial insemination must be used in conjunction with an official serological test, and classification will be based on the maximum titer of either test.
(10) Any animal found to be infected through isolation of field strain Brucella abortus must be classed as a reactor.
(11) Evaluation of titer responses for all cattle and bison shall be the responsibility of a designated epidemiologist, who takes into consideration animal and herd history and other epidemiologic considerations.
(12) Interpretation of market tests. Blood samples collected at slaughter are considered negative if no evidence of agglutination is disclosed on the buffered acidified plate antigen test. Samples showing agglutination are classified negative if the confirmatory standard card test is negative.
Rulemaking Authority Florida Statutes § 585.002(4), 585.08(2) FS. Law Implemented 585.08(1), (3), 585.11(1), (2), 585.155(1), (2), (3), (4) FS. History-New 6-1-92, Amended 7-5-95.