Effects of pesticides upon slug, land snail, annelid, - USC...Once in the laboratory the soil was dried at room temperature, and sieved through a 2-mm sieve. The fraction > 2 mm

$Page 1: Effects of pesticides upon slug, land snail, annelid, - USC...Once in the laboratory the soil was dried at room temperature, and sieved through a 2-mm sieve. The fraction > 2 mm$

Effects of pesticides upon slug, land snail, annelid,

nematode and arthropod populations in horticultural

soils in Galicia

(North-West Spain)

Preliminary results.

Authors:

• Castillejo, J.

• Iglesias, J.

Department of Animal Biology

Faculty of Biology

University of Santiago de Compostela

E-15782 Galica. Spain

OBJETIVES:

Study the effect of

Phasmarabditis nematode strains

on arable soil infaunae

Assess the impact of some

molluscicides on non-target soil

invertebrates

Infaune monitored

MOLLUSCICIDES TESTED:

• Phasmarhabditis

• Metaldehyde

• Cow slurry/Slug Eggs Killer

• Herbicides/Slug Eggs Killer

Round Worms

Slugs

Snails

Earthworms

Springtails Coleoptera

NO TILLED parcel for 5 years before trials

• Ecosystem soil no disturbed

• Animal colony soil established

First Parcel

50 cm depth TILLED parcel, ploughed yearly

• Destroyed ecosystem soil

• New animal soil reorganitation

• New soil colonisation

Second Parcel

1.4 m

1.4 m

2 m2

PARCEL miniPLOTS

• Six replicates for each crops/test

• Six controls plots/random selection

• Monthly sampling

2.6 m

2.6 m

Pontecesures O Grove Santiago

TEMPERATURE (ºC) 13-14 >14 12-13

RAINFALL (mm) 1300-1500 1100-1300 1200-1300

MOISTURE (%) 23.85 34.68 42.44

POROSITY (%) 59.53 77.20 71.89

AIR CONTENT (%) 35.68 42.52 29.45

FRACTION > 2 mm (g) 00.20 45.45 02.49

GROSS SAND (%) 51.18 59.91 36.42

THIN SAND (%) 41.37 20.98 45.87

SLIME (%) 02.80 16.42 11.23

CLAY (%) 04.64 00.267 06.46

pH 05.4 07.4 05.1

pK 04.9 07.2 04.2

pV 05.7 06.5 04.9

C/N Relation 09.58 18.66 08.82

C (meq/100 g) 01.15 03.95 02.56

N (meq/100 g) 00.12 00.21 00.29

Na (meq/100 g) 00.00 00.55 00.25

K (meq/100 g) 00.13 00.13 00.20

Ca (meq/100 g) 01.51 20.90 01.5

Mg (meq/100 g) 00.77 00.67 00.7

Al (meq/100 g) 00.25 00.25 00.91

Soil analyses

Edaphic Analyses For the granulometric

analysis, we took 1kg soil

sample. Once in the

laboratory the soil was

dried at room

temperature, and sieved

through a 2-mm sieve.

The fraction > 2 mm

constitute the gravel.

With the fraction < 2 mm

we did the following

analysis: one part was

used for the

granulometries, and we

separated the thick sand

(2-0.2 mm); the thin

sand, big lime (0.05-0.02)

and clay (<0.0002 mm).

The other part is used for

chemical analysis: pH,

total N2, Organic carbon,

Relation Carbon/Nitrogen

and Oxidable organic

matter. Same soil was

used to make analysis of

moisture, air content and

porosity.

SEASONAL HORTICULTURAL CROPS

CHARDS (Beta vulgaris) LETTUCE (Lactuca sativa)

BRUSSELS SPROUTS CAULIFLOWER

AREA FOR INFAUNAE SAMPLING

14 m2

2 m2

DIGGING for Earthworms, Lands Snails and Macroarthropods

20 cm deep

25 cm

SAMPLING MONTHLY

Surface: 0.0625 x 6 = 0.4 m2

Volumen: 0.0125 m3

SEASONS AND MONTHS

Spring: March, April and May

Summer: June, July and August

Autumn: September, October and November

Winter: December, January, February

Sieves Column

SEARCHING

Inside Soil Samples

for Lands Snails,

Earthworms and Arthropods

PUNCHING for

nematode, mite

and springtail soil

samples

TESTING COMPOUNDS

on NO TILLED arable soil for 5 years

MOLLUSCICIDES SPRAYED ON miniPLOTS before seasonal planting

• Phasmarhabditis nematode strains

• Metaldehyde pellets

• Cow slurry/Slug Eggs Killer

METALDEHYDE: 3 g/m2 of mini-pellets

(Caraquim, Massó Chemical Industries)

containing 5% Metaldehyde, broadcasted evenly

over the surface of the central area of the plots on

the same day of planting

Phasmarhabditis nematode strains:

3 x 105 nematodes m-2 (Nemaslug)

COW SLURRY: 2 litres m-2 of bovine-manure

(equivalent to 20 m3 / hectare)

TARGETS:

• Earthworms

• Slugs and Snails

• Arthropods

First Parcel

Data will be analysed using SPSSx for significance

Before starting experiments the soil fauna was assessed on every plot

and its spatial distribution was homogeneous

Periphery • Phasmarhabditis

Centre •Phasmarhabditis

•Metaldehyde

•Cow slurry + Phasmarhabditis



MORFOLOGÍA EXTERNA DE LAS LOMBRICES

FIGURA 185. Anélidos oligoquetos. Morfología externa de Lumbricus

MORFOLOGÍA EXTERNA

1. Animales terrestres o marinos

2. Alargados con metamería Homónoma

3. No tienen podios, las sedas se insertan

directamente en el cuerpo

4. Tamaño: Chaetogaster mide 0,5 mm y

Megascolex (Megascolécido) mide 3 m

5. Cuerpo formado por anillos que se repiten,

dividido en 3 zona:

Protomio (no segmento)

Metastomio

Pigidio (no segemento)

6. Poros genitales ♀♀: lumbrícidos seg. 14

7. Poros geniales ♂♂: segmento 15 (posición varía

con las especies)

8. Poros de las espermatecas

9. Poros de los nefridios: un par por segmento

10. Poros dorsales

11. Clitelo: espesamiento glandular

Forma anular

Forma de herradura

Tubérculos pubertarios

Formación del capullo

12. Quetas por anillo (8)

DISPOSICIÓN Y ESTRUCTURA DE LAS QUETAS

Esophagus

Pharynx

Gizzard Crop Calciferous

glands

Intestine

Digestive system


1. CÓPULA: surcos o canales

exteriores

2. Clitelo con glándulas de la

albúmina producen capullo.

Fecundación dentro del

capullo: capullo + óvulo +

esperma = se cierra el

limoncillo, puede haber

hasta 20 huevos dentro del

capullo

3. El desarrollo es directo, no

hay metamorfosis, ni larvas

de vida libre, del capullo

sale una lombriz igual que

el adulto

CÓPULA

El capullo de lombriz

DISTRIBUCIÓN VERTICAL DE LAS LOMBRICES

Epigeas

Anécicas

Endogeas

Epigeas

Endogeas

Anécicas

SPECIES of EARTHWORMS FOUND

•Lumbricus terrestris

•Lumbricus castaneus

•Lumbricus rubelus

•Lumbricus friendi

•Allobophora oliveirae

•Allobophora caliginosa

•Allobophora parva

•Eisenia eiseni

•Eisenia veneta

•Eisenia foetida

•Dendrobaena octaedra

•Dendrobaena madeirensis

•Dendrobaena rubida

TYPE OF TERRESTRIAL ANNELID WORMS

•EPIGEIC: Top soil species

•ENDOGEIC: Inside soil species

•ANECIC: Vertical burrow soil species

LAND SNAIL SPECIES MONITORED •Deroceras reticulatum

•Arion intermedius

•Arion ater

•Ponentina ponentina

•Oxychillus sp.

*Number of Oligochaete worms

for 1.2 m2/ for Season

Phasmarhabditis vs Earthworms*

0

10

20

30

40

50

60

70

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 14 49 66 25 0 24

Nema Centre 11 45 37 5 1 40

Nema Periph 4 37 38 21 8 24

Nema Once 6 35 23 5 1 8

Nema+Cow 20 39 26 7 3 16

Summer 99 Autumn 99 Winter 99 Spring 00 Summer 00 Autumn 00

Periphery

Centre

• Six replicates for each crops/test

• Six controls plots

• Monthly sampling

SAMPLING MONTHLY

Surface: 0.0625 x 6 = 0.4 m2

Volumen: 0.0125 m3

SEASONS AND MONTHS

Spring: March, April and May

Summer: June, July and August

Autumn: September, October and November

Winter: December, January, February

*Lumbricus juveniles and adults for 1.2 m2

Phasmarhabditis vs *Lumbricus Earthworms

0

10

20

30

40

50

60

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 4 44 57 21 0 9

Nema Centre 4 39 21 3 0 32

Nema Periph 2 27 21 17 6 18

Nema Once 4 10 14 2 1 1

Nema+Cow 1 14 12 6 0 4


•Lumbricus terrestris

•Lumbricus castaneus

•Lumbricus rubelus

•Lumbricus friendi

• EPIGEIC: Top soil species

• ANECIC: Vertical burrow soil species

Phasmarhabditis vs Lumbricus friendi anecic worms

0

10

20

30

40

50

60

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 3 41 49 21 0 6

Nema Centre 2 5 5 2 0 0

Nema Periph 0 5 7 5 0 0

Nema Once 0 5 11 2 0 1

Nema+Cow 1 12 6 4 0 2


* for 1.2 m2





Periphery

Centre

Phasmarhabditis vs Lumbricus rubellus epigeic worms

0

2

4

6

8

10

12

14

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 1 3 8 0 0 3



Nema Once 4 5 3 0 1 0

Nema+Cow 0 2 4 2 0 2


* for 1.2 m2 •EPIGEIC: Top soil species




* Number of Oligochaete for 1.2 m2

Phasmarhabditis vs Lumbricus juveniles Earthworms

0

5

10

15

20

25

30

35

40

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 2 14 11 4 0 22


Nema Periph 2 22 7 12 4 17

Nema Once 1 16 12 2 2 23

Nema+Cow 1 27 10 2 1 13



* Slugs for 1.2 m2

Phasmarhabditis vs Deroceras reticulatum Slugs

0

20

40

60

80

100

120

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 31 25 65 89 31 7

Nema Centre 56 17 66 89 17 14

Nema Periph 36 21 55 75 26 27

Nema Once 38 26 98 92 31 27

Nema+Cow 40 25 84 90 19 12


Periphery

Centre


Phasmarhabditis vs Arion intermedius Slugs

0

5

10

15

20

25

30

35

40

45

50

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 6 10 27 18 7 7

Nema Centre 8 6 35 24 6 10

Nema Periph 5 12 39 22 1 7

Nema Once 22 8 47 24 8 5

Nema+Cow 13 4 36 15 5 5


* Slugs for 1.2 m2

Phasmarhabditis vs Arion ater Slugs

0

1

2

3

4

5

6

7

8

9

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 0 1 8 0 0 1



Nema Once 0 3 4 6 0 0

Nema+Cow 0 6 8 5 0 0


* for 1.2 m2

Phasmarhabditis vs Oxychilus Snails

0

5

10

15

20

25

30

35

40

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 1 11 24 6 2 4

Nema Centre 1 17 34 11 1 3

Nema Periph 1 13 28 10 1 2

Nema Once 0 12 14 12 1 6

Nema+Cow 4 6 25 4 1 10


Phasmarhabditis vs Ponentina Snails

0

1

2

3

4

5

6

7

Control

Nema Centre

Nema Periph

Nema Once

Nema+Cow

Control 3 4 2 6 2 1



Nema Once 5 0 0 2 0 2

Nema+Cow 3 2 0 3 0 1


*Number of Oligochaete worms for 1.2 m2

Lumbricus, Allobophora, Eisenia, Dendrobaena Genera Periphery

Centre

Metaldehyde and Cow Slurry vs Earthworms

0

10

20

30

40

50

60

70

Control

Meta Centre

Nema+Cow

Control 14 49 66 25 0 24

Meta Centre 1 28 27 5 0 7

Nema+Cow 20 39 26 7 3 16


Collateral effect on bordering areas




Metaldehyde, Cow slurry vs *Lumbricus Earthworms

0

10

20

30

40

50

60

Control

Meta Centre

Nema+Cow

Control 4 44 57 21 0 9


Nema+Cow 1 14 12 6 0 4


*Lumbricus juveniles and adults for 1.2 m2





Metaldehyde, Cow slurry vs Lumbricus friendi anecic worms

0

10

20

30

40

50

60

Control

Meta Centre

Nema+Cow

Control 3 41 49 21 0 6


Nema+Cow 1 12 6 4 0 2


* for 1.2 m2





Periphery

Centre

Metaldehyde, Cow slurry vs Lumbricus rubellus epigeic worms

0

2

4

6

8

10

12

14

16

18

Control

Meta Centre

Nema+Cow

Control 1 3 8 0 0 3


Nema+Cow 0 2 4 2 0 2


* for 1.2 m2


Periphery

Centre

0

5

10

15

20

25

30

Control

Meta Centre

Nema+Cow

Control 2 14 11 4 0 22


Nema+Cow 1 27 10 2 1 13


Metaldehide, Cow Slurry vs Lumbricus juveniles Earthworms

* for 1.2 m2




Metaldehyde, Cow Slurry vs Deroceras reticulatum slugs

0

10

20

30

40

50

60

70

80

90

100

Control

Meta Centre

Nema+Cow

Control 31 25 65 89 31 7

Meta Centre 55 16 72 73 17 8

Nema+Cow 40 25 84 90 19 12


* for 1.2 m2 Periphery

Centre


Metladehyde, Cow Slurry vs Arion intermedius Slugs

0

10

20

30

40

50

60

Control

Meta Centre

Nema+Cow

Control 6 10 27 18 7 7

Meta Centre 14 9 54 44 6 4

Nema+Cow 13 4 36 15 5 5


* for 1.2 m2


Metaldehyde, Cow Slurry vs Arion ater Slugs

0

1

2

3

4

5

6

7

8

9

Control

Meta Centre

Nema+Cow

Control 0 1 8 0 0 1


Nema+Cow 0 6 8 5 0 0


* for 1.2 m2


Metaldehyde, Cow Slurry vs Oxychilus Snails

0

5

10

15

20

25

30

35

Control

Meta Centre

Nema+Cow

Control 1 11 24 6 2 4

Meta Centre 2 11 30 5 0 14

Nema+Cow 4 6 25 4 1 10


* for 1.2 m2


Metaldehyde, Cow Slurry vs Ponentina Snails

0

1

2

3

4

5

6

7

8

9

10

Control

Meta Centre

Nema+Cow

Control 3 4 2 6 2 1


Nema+Cow 3 2 0 3 0 1


* for 1.2 m2


Phasmarhabditis, Cow Slurry, Metaldehyde vs Coleoptera

0

10

20

30

40

50

60

70

80

Control 43 16 29 27 22 19

Nema Centre 45 28 21 25 9 5

Nema Periph 69 24 26 18 20 3

Nema+Cow 30 35 21 30 11 6

Nema Once 62 25 30 20 19 13

Metaldehide 37 15 21 16 14 15


Periphery

Centre


TESTING COMPOUNDS ON ARABLE Soil, 50 cm depth ploughed yearly before trials

MOLLUSCICIDES SPRAYED ON miniPLOTS • Phasmarhabditis nematode strains

• Metaldehyde pellets

• Herbicide Centrol H, Ioxynil active principle/Slug Eggs Killer

METALDEHYDE: 3 g/m2 of mini-pellets (Caraquim,

Massó Chemical Industries) containing 5%

Metaldehyde, broadcasted evenly over the surface of

the central area of the plots on the same day of

planting

Phasmarhabditis nematode strains:

3 x 105 nematodes m-2 (Nemaslug)

HERBICIDE: Comercial name Certrol H

Composition Ioxynil 12% ; MECOPROP 36% .

Manufacture Comercial Quimica MASSO S.A

Activity Herbicide

Second Parcel




Periphery (Nothing)

Centre

Phasmarhabditis

Metaldehyde

Ioxynil (Herbicide)

Phasmarhabditis + Ioxynil

Phasmarhabditis + Metaldehyde

14 m2

2 m2


* Earthworms for 1.2 m2

Phasmarhabditis vs Oligochaete Worms*

0

1

2

3

4

5

6

7

8

Control

Nematode

Nema+Ioxy

Nema+Meta

Control 4 3 1 7

Nematode 3 2 1 3

Nema+Ioxy 0 1 3 6

Nema+Meta 1 0 3 5

Winter 99 Spring 00 Summer 00 Autumn 00

Periphery

Centre


EARTHWORMS

• Allobophora oliveirae (Epig.)

• Allobophora caliginosa (Endog.)

• Dendrobaena octaedra (Epig.)

Phasmarhabditis vs Deroceras reticulatum Slugs

0

2

4

6

8

10

12

Control

Nematode

Nema+Ioxy

Nema+Meta

Control 0 1 1 8

Nematode 0 2 1 11

Nema+Ioxy 0 4 1 4

Nema+Meta 0 1 3 4


* for 1.2 m2


LAND SNAIL

• Deroceras reticulatum

• Arion intermedius

• Arion ater

• Cepaea nemoralis

Phasmarhabditis vs Coleoptera Arthropods*

0

5

10

15

20

25

30

35 Control

Nematode

Nema+Ioxy

Nema+Meta

Control 1 11 30 9

Nematode 3 23 32 15

Nema+Ioxy 2 26 26 19

Nema+Meta 1 18 22 17


O.COLEOPTERA

G. Harpalus

G. Anisodactylus

G. Amara

G. Acupalpus

G. Poecilus

G. Calathus

G. Claenius

G. Xantolinus

*Number of Coleoptora for 1.2 m2


Metaldehyde, Ioxynil vs Oligochaete Worms

0

1

2

3

4

5

6

7

8

Control

Metaldehide

Ioxynil

Nema+Ioxy

Nema+Meta

Control 4 3 1 7

Metaldehide 2 2 5 5

Ioxynil 2 2 3 7

Nema+Ioxy 0 1 3 6

Nema+Meta 1 0 3 5


Periphery

Centre


* Earthworms for 1.2 m2

EARTHWORMS

Allobophora oliveirae (Epig.)

• Allobophora caliginosa (Endog.)

• Dendrobaena octaedra (Epig.)

Metaldehyde, Ioxynil vs Deroceras reticulatum Slugs

0

1

2

3

4

5

6

7

8

9

Control

Metaldehide

Ioxynil

Nema+Ioxy

Nema+Meta

Control 0 1 1 8

Metaldehide 0 0 0 5

Ioxynil 0 1 0 5

Nema+Ioxy 0 4 1 4

Nema+Meta 0 1 3 4


* for 1.2 m2


Metaldehyde, Ioxynil vs Coloptera Arthropods

0

10

20

30

40

50 Control

Metaldehide

Ioxynil

Nema+Ioxy

Nema+Meta

Control 1 11 30 9

Metaldehide 4 22 19 7

Ioxynil 4 25 46 10

Nema+Ioxy 2 26 26 19

Nema+Meta 1 18 22 17


* for 1.2 m2


Impact on Crops

Soil Infaunae

Impact on Adjacent Areas

Soil Infaunae

Oligo. Slugs Snails Arthr. Oligo. Slugs Snails Arthr.

Phasmarhabditis Yes No No No Yes No No No

Phasmarh. Once Yes –— –— No Yes No No No

Cow Slurry Yes –— –— No Yes No No No

Herbicide: Ioxynil –— –— –— –— –— –— –— –—

Metaldehyde Yes –— –— No Yes No No No

–— In Process

PRELIMINARY CONCLUSIONS. During this trial was observed:

•1.- Phasmarhabditis hermaphrodita strains scattered on arable soils at 3 x 105 strain-nematodes

for m2 reduce the population of earthworms. The effect is more significant on anecic Oligochaetes

than on epiegeic species.

•2.- Phasmarhabditis hermaphrodita strains used a molluscicide dose on horticultural crops have

collateral effect on bordering areas earthworm populations.

•3.- The effect of Phasmarhabditis nematode strains on earthworms could persist for more than two

years on arable soils and bordering areas.

•4.- Phasmarhabditis strains haven't any effect on Slug, Snail, Coleoptera and Crane fly populations,

neither in direct application zones nor in bordering areas.

•5.- Metaldehyde mini-pellets, used as molluscicide (3 g/m2) has similar effects on anecic

earthworms than Phasmarhabditis strains in direct application areas and adjacent zones.

•6.- Metaldehyde applied on arable soils in molluscicide doses haven't any collateral effects on

adjacent areas slug, snail and Coleoptera populations.

•7.- Cow slurry applied on arable soil as nitrogen fertilizer (20 m3 / hectare) have collateral effects on

adjacent areas anecic earthworm populations, and no effect on others animals popultions monitored.

•8.- On Ioxynil (Herbicide) effects upon slug eggs we haven't enough field trials, we are working on.

Tested on standard artificial soil work destroying slug eggs.

•9.- The molluscicides compounds tested on 50 cm-depth-ploughed-parcel were deterred by the

ploughing disturbance effect on infauna structure populations caused by farming. Perhaps, one year

period could be necessary for reorganise infauna ecosystem parcels after deep ploughing farm works.




Documents

Effects of pesticides upon slug, land snail, annelid, - USC...Once in the laboratory the soil was dried at room temperature, and sieved through a 2-mm sieve. The fraction > 2 mm