SANS 6095-2009 Stock remedies - Efficacy of chemicals against adult ticks《常备药物 抗扁虱化学品的药效》.pdf

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3、damage whatsoever than may result from the use of this material or the information contain therein, irrespective of the cause and quantum thereof. ISBN 978-0-626-22718-0 SANS 6095:2009Edition 2SOUTH AFRICAN NATIONAL STANDARD Stock remedies Efficacy of chemicals against adult ticks Published by SABS

4、Standards Division 1 Dr Lategan Road Groenkloof Private Bag X191 Pretoria 0001Tel: +27 12 428 7911 Fax: +27 12 344 1568 www.sabs.co.za SABS SANS 6095:2009 Edition 2 Table of changes Change No. Date Scope Foreword This South African standard was approved by National Committee SABS SC 1028E, Pesticide

5、s Livestock remedies, in accordance with procedures of the SABS Standards Division, in compliance with annex 3 of the WTO/TBT agreement. This document was published in July 2009. This document supersedes SABS SM 1095:1987 (first edition). A reference is made in 2.12 to a reference chemical that is r

6、egistered as a stock remedy under “the current relevant national legislation”. In South Africa this means the Fertilizers, Farm Feeds, Agricultural Remedies and Stock Remedies Act, 1947 (Act No. 36 of 1947). SANS 6095:2009 Edition 2 1 Contents Page Foreword 1 Scope 3 2 Equipment and materials 3 3 Te

7、st ticks . 4 4 Procedure . 4 5 Calculations . 7 Bibliography . 8 SANS 6095:2009 Edition 2 2 This page is intentionally left blank SANS 6095:2009 Edition 2 3 Stock remedies Efficacy of chemicals against adult ticks 1 Scope This standard specifies a method for the determination of the efficacy of a ch

8、emical compound (at various concentrations) against engorged adult female ticks, as indicated by their reproductive ability after treatment. 2 Equipment and materials 2.1 Two Incubators, maintained at a temperature of 27 C 1 C and at a relative humidity of 85 % 5 %. 2.2 Stoppered measuring cylinders

9、, of capacity 100 mL. 2.3 Erlenmeyer flasks, of capacity 100 mL and provided with foam plastics stoppers that are protected by tissue paper or cloth squares. 2.4 Glass specimen tubes, of capacity 25 mL and provided with plastics stoppers. 2.5 Test panels, at least three glass panels, each of approxi

10、mate size 125 mm 125 mm and with strips of double-sided adhesive tape pasted in parallel rows onto one side. 2.6 Plastics dishes, suitable plastics dishes for use instead of test panels when carrying out the test with Amblyomma sp. 2.7 Aluminium foil dishes,of diameter approximately 150 mm and of de

11、pth approximately 18 mm. NOTE Alcan style aluminium foil dishes have been found suitable. 2.8 Filter paper,of diameter 150 mm. NOTE Whatman No. 1 filter paper has been found suitable. 2.9 Stainless steel spatula. 2.10 Stopwatch. 2.11 Acetone, of analytical reagent grade or other organic solvent suit

12、able for cleaning purposes. SANS 6095:2009 Edition 2 4 2.12 Reference chemical, that is preferably registered as a stock remedy under the current relevant national legislation (see foreword), the active ingredient of which preferably belongs to the same chemical group as or has a mode of action simi

13、lar to that of the test chemical. 2.13 Test chemical, of sufficient quantity to provide at least 100 mL of each of the six dilutions referred to in 4.1.3. 3 Test ticks 3.1 Select at least 80 fully engorged adult female ticks of uniform size and free from visible abnormalities. NOTE It is essential t

14、hat all ticks used in the test originate from the same batch. 3.2 Wash the ticks in water and dry them in an air current at room temperature. 3.3 Divide the ticks into groups according to size and then randomly allocate 10 ticks to each treatment group to be used (i.e. for treatment with the control

15、 solution, the reference chemical solution and each concentration of the diluted test chemical). Place the groups of ticks in suitable separate containers in which the ticks will not be overcrowded. 3.4 Record the mass of each treatment group. 4 Procedure 4.1 Preparation of equipment and materials 4

16、.1.1 Control solution Use the solvent (or mixture of solvents) that is recommended by the sponsor for the dilution of the test chemical. 4.1.2 Reference chemical Using a suitable solvent, dilute the reference chemical to between 60 % and 80 % of the recommended concentration, or to a concentration t

17、hat has in a similar test been found to be less than 100 % effective but more than 50 % effective against the same tick species. Pour the dilution into a stoppered measuring cylinder and shake well. 4.1.3 Test chemical Prepare a series of six dilutions (X, X/2, X/4, X/8, etc.) of the test chemical i

18、n measuring cylinders and shake well. If the material is a solid, dissolve it in acetone or other suitable organic solvent and dilute with water. If it is a liquid, dilute with the organic solvent or with water or with both. 4.2 Exposure 4.2.1 Application of materials 4.2.1.1 Transfer the ticks in t

19、he control group to a glass specimen tube. Adequately cover the ticks with the control solution, stopper the tube and shake gently. SANS 6095:2009 Edition 2 5 4.2.1.2 Repeat the procedure on the ticks in the reference chemical treatment group, then on the ticks in the test chemical treatment groups

20、(in ascending order of concentration of the test chemical), shaking each solution before immersing the ticks. NOTE Each consecutive treatment group should be immersed in the applicable solution at exactly half-minute intervals so that all the treatment groups are immersed within 4 min. A stopwatch s

21、hould be used to ensure proper timing. 4.2.1.3 During the remainder of the 10 min period, gently shake the stoppered specimen tubes that contain the ticks several times. 4.2.2 Transfer of ticks 4.2.2.1 Exactly 10 min after immersion of the ticks in the control group, decant the liquid off this group

22、 of ticks, and invert the ticks onto a clean, dry piece of filter paper in an aluminium foil dish. Place all the ticks on their ventral sides, each on a dry portion of the filter paper. 4.2.2.2 So repeat the procedure given in 4.2.2.1 (at exactly half-minute intervals) on the ticks in the reference

23、chemical group and then on the ticks in the test chemical group (in ascending order of concentration of the test chemical) that the entire transfer procedure is completed within 4 min. 4.2.2.3 Dry the ticks in an air current at room temperature for at least 1 h. 4.2.2.4 Paste the ticks of the contro

24、l group onto the adhesive strips on a test panel with their ventral sides facing upwards, keeping their capitula clear of the tape, or in the case of Amblyomma sp., place all the ticks in the group in a plastics dish. Place the panel or the plastics dish (as relevant) in the incubator reserved for c

25、ontrol specimens. 4.2.2.5 Repeat the procedure given in 4.2.2.4 on the ticks in the reference chemical group and on those in the test chemical group (in ascending order of concentration of the test chemical), placing the panels or plastics dishes, as relevant, in the incubator reserved for treated s

26、pecimens. NOTE Ticks treated with the test chemical of various concentrations may be pasted on the same panel. 4.3 Results NOTE A form such as that given in figure 1 should be used for recording the test results. 4.3.1 Examine the ticks after 7 d and record the number of dead ticks that have become

27、black (i.e. the mortality of which is not treatment related). 4.3.2 Examine the ticks again at the end of the respective egg-laying periods characteristic to the species (three weeks for Boophilus spp., four weeks for Rhipicephalus spp. and seven weeks for Amblyomma sp.) and record the mass of eggs

28、oviposited by each treatment group and then, using the spatula, transfer the eggs to appropriately labelled Erlenmeyer flasks. Stopper the flasks, place them in the relevant incubator and record the time. Discard the female ticks. 4.3.3 Remove the flasks from the incubators at the end of the hatchin

29、g period characteristic to the species (this period is the same as the egg-laying period) and estimate and record the number of larvae that have hatched, using a numerical scale of 0 to 4. 4.3.4 Repeat the procedure given in 3.1 to 4.3.3 (inclusive) three times to obtain four replicates of the test.

30、 SANS 6095:2009 Edition 2 Species: (susceptible/resistant) Number of adults alive after one week (natural mortality) Mass of females mg Egg laying ability (mass in mg) Egg hatchability (numerical rating) aReplicates (out of) Replicates Replicates Replicates Sample code No. Concentration mg/L 1 2 3 4

31、 Total1 2 3 4 Total1 2 3 4 Total1 2 3 4 MeanControl% NOTE 1 Egg laying assessment: Boophilus ticks: three weeks after treatment. Rhipicephalus ticks: four weeks after treatment. Amblyomma ticks: seven weeks after treatment. NOTE 2 Egg hatchability estimation: Same periods as indicated in NOTE 1 afte

32、r egg laying has ceased. aNumerical rating of egg hatchability: 0 = 0 % 1 = 25 % 50 % 75 % to 100 % Figure 1 Form for recording laboratory test results for engorged adult female ticks 6 SANS 6095:2009 Edition 2 7 5 Calculations 5.1 Calculate the reproductive estimate (R) of each treatment group as f

33、ollows: 124=mNHRmSwhere m1 is the mass of eggs per treatment group, in milligrams; N is the number of ticks per treatment group; H is the hatchability of eggs, in a scale of 0 to 4; m2 is the mass of females per treatment group, in milligrams; S is the number of survivors after seven. 5.2 Separately

34、 calculate the percentage control (C) obtained with the reference chemical and with the different strengths of the test chemical, expressed as a percentage, as follows: yyx100=1RRCRwhere Ry is the reproductive estimate of ticks treated with control solution; Rx is the reproductive estimate of ticks

35、treated with reference chemical or test chemical. SANS 6095:2009 Edition 2 8 Bibliography Drummond, R.O., et al. 1966. Boophilus annulatus and B. microplus: Laboratory test of insecticides. Journal of Economic Entomology, 66:130-133. Holdsworth, P.A., et al. 2006. World Association for the Advanceme

36、nt of Veterinary Parasitology (W.A.A.V.P.) Guidelines for evaluating the efficacy of acaricides against ticks (Ixodidae) on ruminants. Veterinary Parasitology, 136:29-43. SABS SABS Standards Division The objective of the SABS Standards Division is to develop, promote and maintain South African Natio

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