NY/T 3092-2017Chemical pesticide - Guideline for honeybee semi-field test (English PDF)

5.1 Materials and conditions

5.1.1 Trial bees

The bee type for the trial is a representative local bee species which has a definite

strain. The bee colony for the trial shall come from the same breeding strain; the

original and quality shall be reliable. Bee colony having any visible biological pest or

disease, shall not be used for the trial. Bee colony shall not be treated with pesticides

within 4 weeks before it is used for the trial. Queen bees shall be hatched from the

sister queens of the same batch which do mating flight at the same time. Only the

healthy bee queens with strong fecundity, which are newly-bred before the trial, can

be used for the trial. Each colony shall contain 3000 ~ 5000 worker bees and 3 full

frames at least. But it can be adjusted in accordance with the objectives of the trial.

Each colony shall contain all brood stages and an appropriate amount of stores of

pollen/nectar for the development of brood. The colony for the trial shall be in the prime

breeding period or active foraging period.

5.1.2 Test crops

Select crops which are highly attractive to bees, such as rape, cotton and so on. When

the test product is a systemic seed treatment agent or granula, the realistic target crops

of nectar/pollen source recommended for it shall be used.

5.1.3 Test products

Formulated products of pesticides shall be used.

5.1.4 Major equipment and facilities

5.1.4.1 Beehives

Local commonly-used beehives need to be used; or they are customized as required

by the trial.

5.1.4.2 Dead-bee traps

In order to collect dead bees, dead-bee traps shall be installed at the front end of the

entrance of each beehive for the trial, at least 3 d before the investigation of honeybee

mortality. Dead-bee traps shall be made of nonirritant, odorless materials such as

distance between the boundary of the whole trial area and the surrounding field shall

be more than 3 m. There shall be not other agricultural activities than the trial, such as

fertilizer application and control of plant diseases and insect pests; it shall be ensured

that the fertilizer used, pesticide products, their operating process, etc. will not exert

any effect on the results of the trial.

5.2.1.2 Monitoring stage

A site with abundant nectar source (such as wild flowers) is selected as the feeding

and monitoring points of the bee colony at the pre-exposure stage and post-exposure

stage; the distance between the monitoring point and the test field shall be greater than

5.2.2 Test duration

The test duration is decided in accordance with the development period of test crops,

the pesticide application period of control target and the objectives of the trial.

5.2.3 Management of test colony

In accordance with the investigation results of all brood state proportions of the colony

before exposure, coordinate and distribute the colony between all treatments. The time

of migration of the test colony can be the dusk or evening when the colony finishes the

flight activities of the day, or the morning before honeybees start flight activities. When

the colony is kept in the test cage, one water source free of contamination needs to be

provided for honeybees (it is preferably prepared independently outside the beehive

or added directly to the feed bunk inside the beehive if necessary). When the

nectar/pollen of the test crops is not abundant, the colony shall be fed with an

appropriate amount of sugar (honey) water and/or pollen, but in order to ensure the

activeness of the foraging activities of honeybees, they shall not be fed excessively.

When the colony is kept at the monitoring point, feeding management can be done by

following the experiences of the local bee keepers, but during the whole trial process,

the following operation shall not be done for the test colony.

a) using pesticides toxic to queen bees or worker bees to control plant diseases and

insect pests of honeybees;

b) switching frames between different colonies or other behaviors, which will affect

the structure and quantity of colonies.

monitoring point, at least continue to observe for 3 weeks, to ensure the evaluation

cycle after the end of exposure includes one development period of worker bee broods

at least.

5.2.5 Pesticide trial

5.2.5.1 Pesticide application method

The application shall be carried out using the application method recommended for the

test plans or test products, according to the local good agricultural practices (GAP). If

the test products need to be used after preparation, they shall be used immediately

after preparation.

5.2.5.2 Pesticide application appliances

Select frequently-used appliances in production; record all the information of the

appliances used, including type, description, model number and operating conditions

(working pressure and orifice diameter). Before each use, calibrate the pesticide

application appliances and calculate their application rate to decide a method for

uniform application for a plot. If the seeding equipment is used, the equipment shall be

calibrated to seed uniformly, quantitatively.

5.2.5.3 Time and frequency of pesticide application

The time and frequency of pesticide application shall be in accordance with the

recommended application method for the characteristics of the test pesticides.

5.2.5.3.1 Time of pesticide application

The pesticide application shall be carried out during the daytime when honeybees are

foraging most actively; or it is adjusted as appropriate in accordance with different

objectives of the trial. When the effects of long period residual effect pesticides on

honeybees are evaluated, the time of application shall be at a certain time interval

before the exposure of honeybees, to prevent the effects of pesticides caused by direct

exposure to honeybees. If the effects of risk mitigation measures are evaluated, they

shall be applied before the active flight activities of honeybees. In addition to this,

consideration shall be given to the local general agricultural practices. For the direct

atomization treatment method, try to avoid rainfall before the treatments become dry.

normally, there shall be no rainfall within 2 h after application; the air speed in cages

during the application shall be less than 2 m/s.

5.2.5.3.2 Frequency of pesticide application

Normally, apply once during the flowering period, but full consideration shall be given

to the maximum dose, residue, acute effects and different objectives of trial research

and the recommended pesticide application method, before the frequency of

investigation area shall be fully removed.

5.2.6.1.2 Investigation content

When worker bees are incapable of removing the dead bees inside the beehives

because of a massive mortality of honeybees, it also needs to investigate the dead

bees at the bottom of all test beehives. Under the dry field conditions, it also needs to

record the number of dead bees on the ground (by counting the dead bees on the

screen laid on the ground). In accordance with the differences in test objectives and

crops, part or whole of the ground area can be investigated.

5.2.6.2 Honeybee flight conditions

The flight condition investigation needs to be carried out when honeybees are moved

to cages until the end of the exposure. During the investigation, count the honeybee

number foraging on flowers and flying across the area, within a certain area (e.g. 1 m2)

within a certain time (at least 15 s) or on a certain number of flowers (e.g. 15 flowers).

Select at least 3 observation points randomly for each cage (avoiding the area before

the beehive entrance).

The flight investigation shall be carried out under normal conditions after the flowers of

the crops in cages bloom, at the time when honeybees fly the most actively; evaluation

shall be carried out once at the same time each day, but on key test points, e.g. the

first day of the exposure, the frequency of flight investigation shall be increased, e.g. 1

h, 2 h, 4 h, 6 h and other periods of time after the exposure.

5.2.6.3 Honeybee activities

At the exposure stage of the cage trial, investigate the activities of honeybees on the

crops and around the beehives in the process of assessing the honeybee mortality and

flight conditions. Compared with the blank controls, at least the following activities shall

be observed and recorded.

a) toxic symptoms, such as convulsion, tremble and unbalanced movement;

b) gathering at the entrance of the beehives;

c) aggressivity;

d) hanging bee beard;

e) inertia;

f) flying in high density without falling on the crops;

g) other abnormal behaviors.