1、 Collection of SANS standards in electronic format (PDF) 1. Copyright This standard is available to staff members of companies that have subscribed to the complete collection of SANS standards in accordance with a formal copyright agreement. This document may reside on a CENTRAL FILE SERVER or INTRA
2、NET SYSTEM only. Unless specific permission has been granted, this document MAY NOT be sent or given to staff members from other companies or organizations. Doing so would constitute a VIOLATION of SABS copyright rules. 2. Indemnity The South African Bureau of Standards accepts no liability for any
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 978-0-626-22079-2 SANS 5462:2008Edition 4SOUTH AFRICAN NATIONAL STANDARD Pesticides Rearing and handling of the yellow fever mosquito (Aedes aegypti
4、 (L.) Published by SABS 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 5462:2008 Edition 4 Table of changes Change No. Date Scope Foreword This South African standard was approved by National Committee
5、SABS SC 1028F, Pesticides Laboratory facilities and rearing and handling of test animals, in accordance with procedures of the SABS Standards Division, in compliance with annex 3 of the WTO/TBT agreement. This document was published in December 2008. This document supersedes SABS SM 462:1977 (second
6、 revision). SANS 5462:2008 Edition 4 1 Pesticides Rearing and handling of the yellow fever mosquito (Aedes aegypti (L.) 1 Scope This standard specifies a method for the rearing and handling of the yellow fever mosquito (Aedes aegypti (L.). 2 Normative references The following referenced documents ar
7、e indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. Information on currently valid national and international standards can be obtained fr
8、om the SABS Standards Division. SANS 5808, Pesticides General conditions and equipment for the rearing and handling of test insects. 3 Rearing conditions Maintain the conditions prescribed in SANS 5808, but keep the rearing room for adults and larvae illuminated for 24 h of the day. 4 Rearing equipm
9、ent 4.1 Aspirator, as described in SANS 5808. 4.2 Cages for adults, mosquito cages as described in SANS 5808. 4.3 Feeding dishes for adults, disposable plastics containers of approximate height 5 mm and diameter 90 mm. 4.4 Guinea pig cage assembly, as described in SANS 5808. 4.5 Oviposition containe
10、rs, plastics containers with wicks as described in SANS 5808, but remove their lids and wicks. 4.6 Larval rearing containers and lids, type B rearing containers and type A rearing container lids as described in SANS 5808. SANS 5462:2008 Edition 4 2 4.7 Sieves 4.7.1 Type A sieve as described in SANS
11、5808 of nominal aperture size 420 m. 4.7.2 Type C sieve as described in SANS 5808. 5 Host animal Guinea pig with a shaven back. 6 Media 6.1 Feeding medium for adults Saturate a fairly large piece of cotton wool with a solution of 5 g/100 mL of brown sugar in distilled water, place it in a feeding di
12、sh (see 4.3), cover the dish with unpolished paper secured with a rubber band, and trim off excess paper close to the feeding dish. 6.2 Oviposition substrate Circle of coarse porosity filter paper, of diameter 150 mm, that is placed as a lining in each oviposition container (see 4.5) and to which 30
13、 mL of distilled water is added. (The females deposit their eggs on the accessible edges of the wet filter paper.) 6.3 Rearing medium for larvae Sift each of the following dry ingredients through the sieve specified in 4.7.1, and mix them together: 4 g fishmeal; 4 g milk powder; 8 g finely ground br
14、own bread; and 1 g brewers yeast. 7 Procedure 7.1 Use the aspirator (see 4.1) connected to a suction apparatus operated at a very low airflow to collect freshly-emerged adult yellow fever mosquitoes (see 7.9) and transfer them to a cage (see 4.2) that contains the oviposition substrate (see 6.2) in
15、an oviposition container (see 4.5) and the feeding medium (see 6.1). Perform this operation on a Monday. 7.2 So place the host animal (see clause 5) in the cage assembly (see 4.4) that only its back is exposed to the mosquitoes (its head projecting through the porthole and being protected by the cag
16、e). The animal is placed in the oviposition cage for 3 h each day from Monday to Friday (inclusive) of every week to allow the adult females to obtain blood meals. 7.3 On Wednesday and Fridays, and on succeeding Mondays, Wednesdays and Fridays, replace the feeding medium with a fresh supply, and rem
17、ove the oviposition substrate with the adhering eggs from the oviposition container and replace it with fresh oviposition substrate. SANS 5462:2008 Edition 4 3 7.4 Keep the removed substrate and the adhering eggs moist for 3 d to ensure complete embryonic development. Allow the substrate to dry, mar
18、k the date of egg-laying on it, and store it in a dry place. 7.5 On a Monday, add to a rearing container (see 4.6) 10 L of distilled water and mix in 0,5 mL of the dry rearing medium (see 6.3). Add a circle of dried substrate (see 7.4) with adherent eggs that are not more than 4 months old. The eggs
19、 will hatch within 24 h. NOTE If a larger colony or larvae of various ages (or both) are required, additional rearing containers may also be prepared on a Wednesday and a Friday, and eggs placed in these. 7.6 On Wednesday, remove the filter paper and again add 0,5 mL to 3,0 mL of the dry rearing med
20、ium to the water in the rearing container, as necessary. NOTE The water should be brown or black. The addition of excess rearing medium will cause the water to turn white. If the larvae are brown, a greater quantity of rearing medium should be added, but if the larvae are black, only a small quantit
21、y of rearing medium is required. 7.7 On Thursday and Friday and the following Monday, add the dry rearing medium to the water in the rearing container, as necessary. 7.8 Whenever the water in the rearing container becomes fouled, transfer, as follows, the larvae (and pupae) to a clean rearing contai
22、ner that contains 10 L of fresh distilled water and 1,0 mL of the dry rearing medium: Pour the fouled water from the original rearing container through the sieve (see 4.7.2) and rinse with distilled water to remove very fine frass. Invert the sieve over a 500 mL beaker that contains distilled water,
23、 and wash the larvae (and pupae) into it using distilled water. Carefully (to prevent the frass from moving from the beaker into the fresh water) immerse the beaker of larvae (and pupae) in the mixture in the clean rearing container. Position a 75 W lamp underneath the container. The larvae (and pup
24、ae) will move away from the light, i.e. from the beaker into the fresh water. After the larvae (and pupae) have evacuated the beaker, remove it and discard the frass. 7.9 Adults will emerge after approximately 10 d to 15 d. Collect adults daily (Mondays to Fridays (inclusive) by sucking them out using the aspirator (see 4.1). 7.10 Rear a new colony, by repeating the procedure in 7.1 to 7.9 (inclusive), but starting a cycle on Wednesdays and on Fridays as well as on Mondays. SABS
