Ascophyllum nodosum, ext.

Toxicity to terrestrial arthropods

Administrative data

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

July - September 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: in accordance with guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: ESCORT I Guidance Document (Barret et al., 1994)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: ESCORT II Guidance Document (Candolfi et al., 2000)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: IOBC Guidelines (Mead-Briggs et al., 2000)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: guideline of the ring testing group (Mead-Briggs et al., 2009)

Deviations:

no

Principles of method if other than guideline:

SOP BT207 , Biotecnologie BT internal procedure.

GLP compliance:

yes (incl. QA statement)

Remarks:

Italian Ministero della Salute

Application method:

spray

Test material

Test material information

Constituent 1

Reference substance name:

Ascophyllum nodosum, ext.

EC Number:

283-907-6

EC Name:

Ascophyllum nodosum, ext.

IUPAC Name:

283-907-6

Details on test material:

- Name of test material : ALGEA FERT SOLID K+
- Substance type: algal extract, intended to be used as agriculture fertilizer
- Physical state: solid, balck micro-flakes
- Lot/batch No.: 00680
- Expiration date of the lot/batch: 09 March 2012
- Stability under test conditions: stable

Test substrate

Details on preparation and application of test substrate:

The applications were performed by a spraying equipment, calibrated to deliver spray at volume rate equivalent to 200 L/ha. Simultaneously other glass plates were sprayed with deionised water for the control and with Perfekthion for the reference substance group.
After the application of the spray solutions, when the treated plates were dried, the exposure units were assembled.
After 48 hours of exposure to the treated glass plates, the surviving females were removed with an aspirator from the exposure cage and they were taken apart. Each female (15 females for the treatment group and 15 females for the control group) was transferred to single fecundity cage, and given 24 hour time period to parasitize aphids. In the fecundity cages there were untreated Aphids infested plants. After 24 hours the females were removed from the fecundity cages and their condition (alive, dead or not recovered) was recorded. The parasitized aphids within the fecundity arenas were left to develop in situ and the number of aphid “mummies” that developed was recorded 17 days later.

Test organisms

Test organisms (species):

Aphidius rhopalosiphi

Animal group:

Hymenoptera (leaf-dwelling parasitic wasp)

Details on test organisms:

TEST ORGANISM
- Common name: Aphidius rhopalosiphi De Stefani Perez
- Source: Biotecnologie BT S.r.l.
- Age at test initiation : Adults
- Disease free: yes
- Kept according to standard practices: yes


- Acclimation period: not relevant

- Feeding: Water/honey solution (During the exposure phase, a piece of a cotton wool soaked with a 1:3 v/v solution of honey and water was inserted into the access hole as a source of food and was re-moistened at least once a day. During the 24 h parasitisation period the parasitoids were fed on the honeydew of the aphids, which were offered for parasitisation).

Study design

Study type:

laboratory study

Limit test:

yes

Total exposure duration:

18 d

Post exposure observation period:

The conditions of the exposed parasitoids were recorded at approximately 2, 24 and 48 hours after their introduction. Counting of parasitized aphids was carried out 17 days after the start of the fecundity test.

Test conditions

Test temperature:

exposure phase: 20.00 – 21.33°C / reproduction phase: 19.33 – 22.00°C

Humidity:

exposure phase: 68.50 – 77.00%

Photoperiod and lighting:

Photoperiod: 16 h light and 8 h darkness
Light Intensity: reproduction phase: 6800 - 9400 lux

Details on test conditions:

TEST SYSTEM
- Test container / cage (material, size): The test system was exposed to the test item on circular glass plates (diameter: 10.00 cm), which were assembled to cages with an aluminium frame (diameter: 10.00 cm; height: 1.50 cm; width: 1.00 cm)after the spray layer has dried.
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 5
- No. of replicates per control : 5
- No. of replicates per reference substance (Perfekthion): 5


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and reproduction performance (at approximately 2, 24 and 48 hours after treatment).

Nominal and measured concentrations:

One single rate equivalent to twice the maximum field application rate multiplied by the MAF (worst case exposure): 21000.00 Product g/ha

Reference substance (positive control):

yes

Remarks:

formulation Perfekthion

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

LR50

Effect conc.:

21 000 g/ha

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Mortality at end of exposure period: very low ( 14%)

Results with reference substance (positive control):

Mortalityat end of exposure period: 96%

Reported statistics and error estimates:

Appropriate statistical methods were used to analyse mortality and fecundity data for significance: Student’ T Test and Anova Manova Test.

Any other information on results incl. tables

Table 1: Mortality at 48 hours

Product	g product/ha	g a.s./ha	% Mortality	% Corrected Mortality
Control	0.00	0.00	6.00	===
ALGEA FERT SOLID K+	21000.00	n.a.	14.00	8.51
Perfekthion	10.00	3.65	96.00	95.74

Table 2. Reproduction performance

Product	g product/ha	g a.s./ha	N° of mummies/♀	±SD	% R
Control	0.00	0.00	9.80	7.29	===
ALGEA FERT SOLID K+	21000.00	n.a.	6.33	6.48	35.37

%R = % reduction in reproduction

During the reproduction performance there was a reduction in reproduction of parasitoids at the tested concentration: this value was 35.37%. The mean for the tested concentration was not different to the control mean at alpha = 0.05, by the Student’s T Test (p = 0.1795).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

In accordance with IOBC/WSPR guidance, the test is valid because the mortality observed in the untreated control group after 48 hours was ≤ 10.00% and the mortality caused by the reference item was ≥ 50%. Moreover the control parasitisation rate (mean) wa

Conclusions:

The results obtained in the trial showed that the test item ALGEA FERT SOLID K+ did not cause a statistically significant mortality of the test organism Aphidius rhopalosiphi De Stefani Perez (Hymenoptera: Braconidae) when applied to glass plates at twice the worst case application rate. At the rate tested, the corrected mortality was < 50%.
The LR50 of ALGEA FERT SOLID K+ calculated at 48 hours after treatment, is higher than 21.00 kg of product/ha.
During the reproduction performance the treated surviving females had a decrease in reproductive capacity equivalent to 35.37%. Than the ER50 value was > 21.00 kg/ha.

Executive summary:

The effects of residues of ALGEA FERT SOLID K+on the mortality and reproduction of the aphid parasitoidAphidius rhopalosiphiDe Stefani Perez (Hymenoptera: Braconidae), were tested with a laboratory study, according to GLP regulations. The test item was sprayed on glass plates with calibrated spray equipment () at one single rate equivalent to twice the maximum field application rate multiplied by the MAF (worst case exposure). The formulation Perfekthion was used as reference substance, whereas deionised water was used as control. Five replicates for each treatment were performed. The sprayed test substances were allowed to dry for about 1 hour, than in each cage 10 adults were introduced.After 48 hours of exposure to the treated glass plates the surviving parasitoids were removed from the exposure cages and the parasitic capacity per female was evaluated in the fecundity test. The conditions of the exposed parasitoids were recorded at approximately 2, 24 and 48 hours after their introduction. Counting of parasitized aphids was carried out 17 days after the start of the fecundity test. The results obtained in the trial showed that the test item ALGEA FERT SOLID K+ did not cause a statistically significant mortality of the test organismAphidius rhopalosiphiDe Stefani Perez (Hymenoptera: Braconidae) when applied to glass plates at twice the worst case application rate. At the rate tested, the corrected mortality was < 50%. The LR₅₀of ALGEA FERT SOLID K+ calculated at 48 hours after treatment, is higher thanof product/ha.

During the reproduction performance the treated surviving females had a decrease in reproductive capacity equivalent to 35.37%. Than the ER₅₀value was > 21.00 kg/ha.