effect of two biological nematicides and aldicarb … · national agrarian university of molina...

NATIONAL AGRARIAN UNIVERSITY OF MOLINA

FACULTY OF AGRONOMY

Effect Of Two Biological Nematicides and Aldicarb in the Control ofMeloidogyne Incognito Chitwood 1949 In the Cultivation of the Olive

Tree in Pisco-Ica

THESIS FOR THE TITLE OF:

AGRICULTURAL ENGINEER

PRESENTED BY

ZOILA ERNESTINA RAVINES ALFARO

2000

INDEX

PAGE

Dedication ...........................................................................................i

Gratefulness .......................................................................................ii

Index .................................................................................................iii

Index of tables...................................................................................iv

Index of figures ..................................................................................v

I INTRODUCTION............................................................................1

1.1 Introduction........................................................................................1

1.2 Objectives ..........................................................................................3

II LITERATURE REVIEW ...............................................................4

2.1 Olive tree cultivation..........................................................................4

2.1.1 Botanical classification ......................................................................4

2.1.2 Olive culture in the world ..................................................................4

2.2 Morphology of the olive tree .............................................................7

2.2.1 General characters..............................................................................7

2.2.2 Root system........................................................................................7

2.2.3 Leaves ................................................................................................7

2.2.4 Flowers...............................................................................................8

2.2.5 Fruit....................................................................................................8

2.3 Yearly vegetative cycle......................................................................8

2.4 Nematodes of the plants.....................................................................9

2.4.1 Effects of Meloidogyne incognito in olive ........................................9

2.5 Characteristic of the root knot nematode .........................................10

2.6 Control of nematodes.......................................................................12

6.1 Control methods for nematodes of the root .....................................12

III MATERIALS AND METHODS ..................................................16

3.1 Geographical location ......................................................................16

3.2 Characteristics of the test area ........................................................16

3.2.1 Temperature .....................................................................................16

3.2.2 Soil ...................................................................................................16

3.2.3 Water................................................................................................16

3.3 Experimental design.........................................................................16

3.3.1 Characteristics of the test area .........................................................17

3.4 Materials and equipment..................................................................17

3.4.1 Vegetative material ..........................................................................17

3.4.1.1 Characteristic of the graft.................................................................18

3.4.1.2 Characteristic of the pattern .............................................................18

3.4.2 Experimental material......................................................................19

3.4.3 Equipment ........................................................................................21

3.4.3.1 Field material ...................................................................................21

3.4.3.2 Laboratory material..........................................................................24

3.4.4 Field work ........................................................................................22

3.4.4.1 Irrigation ..........................................................................................22

3.4.4.2 Fertilization......................................................................................23

3.4.4.3 Disease control.................................................................................23

3.4.5 Methods and procedures ..................................................................23

3.4.5.1 Sampling ..........................................................................................23

3.4.5.2 Application of treatments.................................................................23

5 Experiment variables analyzed ........................................................24

3.5.1 Population of nematodes in the soil .................................................24

3.5.2 Population of nematodes in the root ................................................24

3.5.3 Nodulation index..............................................................................24

3.5.4 Yield and quality of the product ......................................................25

3.5.4.1 Number of fruit per tree ...................................................................25

3.5.4.2 Weight of fruit..................................................................................25

3.5.4.3 Yield per tree ...................................................................................25

IV RESULTS AND DISCUSSION ....................................................25

4.1 Population of M. incognita ..............................................................25

4.1.1 Population of M. incognita in the soil .............................................25

4.1.2 Population of M. incognita in the root.............................................27

4.1.3 Nodulation index ..............................................................................30

4.2 Population of parasitic and non parasitic nematodes .......................32

4.2.1 Population of nematode parasites.....................................................32

4.2.2 Population of non parasitic nematodes.............................................36

4.3 Population of other nematodes found in the soil ..............................40

4.3.1 Population fluctuations of Aphelenchoides in 100 cc of soil ...........40

4.3.2 Population fluctuations of Aphelenchus in 100 cc of soil ................41

4.3.3 Population fluctuations of Ditylenchus in 100 cc of soil .................42

4.3.4 Population fluctuations of Helicotylenchus in 100 cc of soil ..........43

4.3.5 Populations fluctuations of Hemicycliophora if 100 cc of soil........44

4.3.6 Population fluctuations of Pratylenchus in 100 cc of soil ...............46

4.3.7 Population fluctuations of Rotylenchus in 100 cc of soil.................47

3.8 Population fluctuations of Tylenchorhynchus in 100 cc of soil .......48

4.3.9 Population fluctuations of Tylenchus in 100 cc of soil ....................49

4.3.10 Population fluctuations of Xiphinema in 100 cc of soil...................51

4.3.11 Population fluctuations of Criconematidos in 100 cc of soil...........52

4.3.12 Population fluctuations of Dorylaimidos in 100 cc of soil ..............53

4.3.13 Population fluctuations of Mononchidos in 100 cc of soil ..............54

4.3.14 Population fluctuations of Rhabditidos in 100 cc of soil.................55

4.3.15 Population fluctuations of Trichodoridos in 100 cc of soil .............57

4.4 Yield .................................................................................................58

4.4.1 Yield per tree ....................................................................................58

4.4.2 Weight of fruit ..................................................................................60

4.4.3 Number of fruit per tree....................................................................61

V CONCLUSIONS ............................................................................62

VI RECOMMENDATIONS...............................................................64

VII SUMMARY ....................................................................................65

VIII BIBLIOGRAPHY ..........................................................................67

VIII ANNEXES ...................................................................................... ??

INDEX OF TABLES

Page1. World production of olive oil (x1000 t) . . . . . . . . . . . . . . . . . . . . . 5

2. Olive grove acreage in the world by country and most frequent

varieties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

3. Monthly temperatures during the trial (°C) . . . . . . . . . . . . . . . . . . . 18

4. Treatments and application rates . . . . . . . . . . . . . . . . . . . . . . . . . . 19

5. Index of root nodulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

6. Number of juvenile M. incognito in olive trees in 100 cc soil

treated with two biological nematicides, aldicarb and no treatment

in fields infested with root knot nematode (Orchard Alamein-

Pisco-Ica) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

7. Number of juveniles and eggs of M. incognito in 5 g of root of

olive trees treated with two biological nematicides, aldicarb and

no treatment in fields infested with root knot nematode (Huerto

Alamein-Pisco-Ica) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

8. Degree of the nodulation index caused by M. incognito in roots of

tomatoes planted in soil of olive trees treated with two biological

nematicides, aldicarb and no treatment in fields infested with root

knot nematode (Huerto Alamein-Pisco-Ica) . . . . . . . . . . . . . . . . . . 34

9. Number of parasitic nematode in 100 cc soil in olive trees treated

with two biological nematicides, aldicarb and no treatment in

fields infested with nematodes (Orchard Alamein-Pisco-Ica) . . . . . 36

10. Number of parasitic nematode in 5 g of root in olive trees treated


fields infested with nematodes (Orchard Alamein-Pisco-Ica) . . . . . 39

11. Number of non-parasitic nematodes in 100 cc soil in olive trees


in fields infested with nematodes (Orchard Alamein-Pisco-Ica) . . . 41

12. Number of non-parasitic nematodes in 5 g of root in olive trees

treated with two biological nematicides, aldicarb and no treatment 43

Number of Aphelenchoides in 100 cc of soil of olive trees treated


fields infested with nematodes (Orchard Alamein - Pisco-Ica) . . . .

45

14. Number of Aphelenchus in 100 cc of soil of olive trees treated



46

15. Number Ditylenchus in 100 cc of soil of olive trees treated with

two biological nematicides, aldicarb no treatment in fields infested

with nematodes (Orchard Alamein - Pisco-Ica) . . . . . . . . . . . . . . . 47

16. Number Helicotylenchus in 100 cc of soil of olive trees treated



48

17. Number Hemicycliophora in 100 cc of soil of olive trees treated

with two biological nematicides, aldicarb no treatment in fields

infested with nematodes (Orchard Alamein - Pisco-Ica) 49

18. Number Pratylenchus in 100 cc of soil of olive trees treated with

two biological nematicides, aldicarb and no treatment in fields

infested with nematodes (Orchard Alamein - Pisco-Ica) 50 s

19. Number of Rotylenchus in 100 cc of soil of olive trees treated with


infested with nematodes (Huerto Alamein-Pisco-Ica) . . . . . . . . . . . 52

20. Number of Tylenchorhynchuin in 100 cc of soil of olive trees


in fields infested with nematodes (Huerto Alamein-Pisco-Ica). 53

21. Number of Tylenchus in 100 cc of soil of olive trees treated with


infested with nematodes (Huerto Alamein-Pisco-Ica) . . . . . . . . . . . 54

22. Number of Xiphinema in 100 cc of soil of olive trees treated with 55


infested with nematodes (Huerto Alamein-Pisco-Ica) . . . . . . . . . . .

Number of Criconematidos in 100 cc of soil of olive trees treated


fields infested with nematodes (Huerto Alamein-Pisco-Ica) . . . . . .

57

24. Number of Dorylaimidos in 100 cc of soil of olive trees treated



58

25. Number of Mononchidos in 100 cc of soil of olive trees treated



59

26. Number of Rhabditidos in 100 cc of soil of olive trees treated with


infested with nematodes (Huerto Alamein-Pisco-Ica) . . . . . . . . . . .

60

27. Number of Trichodoridos in 100 cc of soil in olive trees treated


fields infested with nematodes (Orchard Alamein -Pisco-Ica) . . . . 61

28. Table of yields of olive trees treated with two biological

nematicides, aldicarb and notreatment in fields infested with root

knot nematode (Orchard Alamein-Pisco-Ica) . . . . . . . . . . . . . . . . . 62

29. Average yield for olive tree treated with two biological


knot nematode (Orchard Alamein-Pisco-Ica) in grams . . . . . . . . . . 63

30. Average weigh of fruit of olive tree treated with two biological



31. Average number of fruit of olive tree treated with two biological



INDEX OF FIGURES

Page1. Number of juvenile of M. incognito in 100 cc soil of olive trees


in fields infested with root knot nematode (Huerto Alamein-Pisco-

Ica) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

2. Number of juvenile and eggs of M. incognito in 5 g of root in

olive trees treated with two biological nematicides, aldicarb and no

treatment in fields infested with root knot nematode (Huerto

Alamein-Pisco-Ica) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

32

3. Nodulation index caused by M. incognito in roots of tomatoes

planted in the soil of olive trees treated with two biological


knot nematode (Orchard Alamein - Pisco-Ica) . . . . . . . . . . . . . . . . 35

4. Number of parasitic nematode in 100 cc soil in olive trees treated


fields infested with nematode (Orchard Alamein-Pisco-Ica) . . . . . .

37

5. Number of parasitic nematodes in 5 g of root in olive trees treated


fields infested with nematode (Orchard Alamein-Pisco-Ica) . . . . . . 39

6. Number of non-parasitic nematodes in 100 cc soil in olive trees


in fields infested with nematode (Orchard Alamein-Pisco-Ica) . . . .

42

7. Number of non-parasitic nematodes in 5 g of root in olive trees


in fields infested with nematode (Orchard Alamein-Pisco-Ica) . . . .

43

8. Number Aphelenchoides in 100 cc of soil of olive trees treated

with two biological nematicides, aldicarb and no treatment in 45

fields infested with nematode (Orchard Alamein -Pisco-Ica) . . . . .

9. Number Aphelenchus in 100 cc of soil of olive trees treated with


infested with nematode (Orchard Alamein - Pisco-Ica) . . . . . . . . . . 46Page

10. Number Ditylenchus in 100 cc of soil of in olive trees treated with


infested with nematode (Orchard Alamein -Pisco-Ica) . . . . . . . . . . 47

11. Number Helicotylenchus in 100 cc of soil of in olive trees treated


fields infested with nematode (Orchard Alamein - Pisco-Ica) . . . . .

48

12. Number Hemicycliophora in 100 cc of soil of olive trees treated



50

13. Number Pratylenchus in 100 cc of soil of olive trees treated with


infested with nematode (Orchard Alamein - Pisco-Ica) . . . . . . . . . .

51

14. Number Rotylenchus in 100 cc of soil of olive trees treated with



52

15. Number Tylenchorhynchus in 100 cc of soil of olive trees treated



53

16. Number Tylenchus in 100 cc of soil of olive trees tried with two


in fields infested with nematode (Huerto Alamein- Pisco-Ica) . . . . 55

17. Number Xiphinema in 100 cc of soil of olive trees tried with two 56

biological nematicides, aldicarb and no treatment in fields infested

with nematode (Orchard Alamein - Pisco-Ica) . . . . . . . . . . . . . . . .

18. Number of Criconematidos in 100 cc soil of olive trees treated



57

19. Number of Dorylaimidos in 100 cc soil of olive trees treated with



58

Number of Mononchidos in 100 cc of soil of olive trees treated



60

21. Number of Rhabditidos in 100 cc of soil of olive trees treated with



61

22. Number of Trichodoridos in 100 cc of soil of olive trees treated



62

23. Average yield of olive tree treated with two biological



24. Average weigh of olive tree fruit treated with two biological



25. Number average of olive tree fruits treated with two biological



1

I. INTRODUCTION AND OBJECTIVES

1.1 INTRODUCTIÓN

The presence of plant parasitic nematode causes more loss in fruit-bearing perennial as

the olive tree, because its above ground symptoms are not specific many times their effects are

attributed to other causes, such as the lack of nutrition, fungus problems, among others. The

economic losses in the production of susceptible cultivars are considered in 25-40% range (20).

Meloidogyne incognito chitwood is a nematode broadly distributed in Peru and in the world and

is of more importance in areas with tropical and subtropical temperatures as in the Peruvian

coast. It attacks more than 700 vegetable species, therefore the use of cultivation rotation

becomes impossible as a control method less so in perennial trees such as those bearing fruit; it is

for this reason that nematicide use is necessary for effective easy control.

The olive (Olea europea) it is a cultivation of relative importance in the South Coast of

Peru and is one of the most susceptible cultivations to the attack of the root knot nematode (M.

incognito) that produces nodules in the roots of the olive tree and in severe attack a causes

decline in the plant. Pratylenchus spp. and Tylenchulus semipenetrans that possibly have some

effect on the production (20, 28) also attack Olive.

Several types of nematode controls are known such as: physical, chemical, biological,

cultural, genetic control. An integrated control that uses two or more control methods (16, 20,

44). The selection of the control method depends mainly on the biology of the nematode, of the

plant host, its value per hectare, the system of diffusion, the usual cultural practices of the area,

the relationships among the ecological factors, and the relative cost of the available methods

(44). The effectiveness of each one of these methods is considered according to the importance

of the cultivation, topography of the land, texture and structures of the soil, climate, irrigation

type, etc. (20).

Until the decade of 1949, the accumulation of the knowledge on the important species of

nematodes for agriculture was slow; during these years the chemical industry introduced

nematicides for the soil, which reduced nematode populations from the soil at a much lower cost

than previously required. Because of the improvements achieved in the growth and yield of

2

plants after the use of soil applied nematicides the importance of nematodes were recognized in

the agriculture and marked the start of the indiscriminate use of this chemical control with these

products (29). The disadvantage of the use of this control method is that after the residual effect

of the product ends, the nematode recovers its initial population level and even overcomes it

(34). This causes phytotoxic problems in the plant, environmental contamination, it affects the

population of beneficent organisms, thereby disrupting the biological balance and with the

excessive increase of parasitic nematode, it is believed to causes a natural disorder. For these

reasons many countries of the world are limiting the use of these chemical products (20).

Scientists have explored different alternatives of combating these pests. Among those

considered, biological control is one of the most effective and inoffensive for the environment

and it is one of those that is acquiring bigger importance in the entire world. In biological control

of nematodes natural enemies such as virus, bacteria, rickettsias, fungus atrapadores,

endoparasites, nematodes, acari, insects, etc. are used, which use benign modes of action to

control parasitic nematode. (20).

The use of biological nematoxins is a form of induced biological control that controls

parasitic nematodes maintaining an ecobalance in the rhizosphere. These products use a

biocontrol action that suppresses the development of eggs and the development of juveniles of

nematodes, they also immobilize the juvenile of eggs newly hatched without damaging the

beneficent fauna (4).

The present work is designed to carry out a comparison between the effects of two

biological nematoxins and aldicarb in the control of M. incognito and on yield of olive. This

work will also offer additional information that will be been able to be used in the control of

nematodes in olive.

3

1.2 OBJETIVES

- To evaluate the efficiency of three nematatoxins on olive (European L. You arise) variety

Sevillana in the reduction of the population of M. incognito and other parasitic

nematodes.

- To evaluate the action of the nematoxins on the populations of non-parasitic nematodes.

- To evaluate the yield and quality of fruit of olive with different treatments of

nematatoxins.

4

II. LITERATURE REVIEW

2.1 THE CULTURE OLIVE

2.1.1 BOTANICAL CLASSIFICATION

Loussert and Brouse mentioned by Guerrero (19), in their book The Olive" (1980),

continue the classification of Emberger (1960) placing it in the Oleaceas family of the

Ligustrales order.

In general Olea está is composed of 30 different species. According to Cifferi and

Breviglieri (1942) mentioned by Guerrero (19), the species Olea europea is divided into three

subspecies:

a) Euromediterranea

Olea europea b) Laperrini

c) Cuspidata

The subspecies Euromediterranea includes the series oleaster and sativa. Oleaster is a

spontaneous form commonly designated acebuche that is distributed throughout the

mediterranean basin. Sativa is a cultivated olive. The subspecies Laperrini exists in northern

Africa as a spontaneous form. The subspecies Cuspidata is found from the northeast of the

Himalayas to Afghanistan. The cultivation of the olive had its origin in Syrian and Iran,

extending toward the West to the Mediterranean. Also, recently it spread to other such areas as

America, Australia, China and South Africa.

2.1.2 THE OLIVE CULTURE IN THE WORLD

The world production of olive oil in the years 1991, 1992 and 1993, according to the

Production Annual of the FAO, can be seen in table 1.

In 1997, Spain by itself accounted for 218 million trees, 30% of the world production of

olive oil and 26% of the production of table olives (3).

The top 4 countries in the world: Spain, Italy, Greece and Turkey account for 79% of the

world production of oil and 55% of the world production of table olive. In descending order for

production of table olives, the main countries plows: Spain, Turkey, USA - California, Greece,

Morocco, Syria, Italy, Argentina, Peru, Portugal, Egypt, and Tunisia (3).

5

Table 1: WORLD PRODUCTION OF OLIVE OIL (X 1000 t)

Countries 1991 1992 1993WORLD 2,007 1,835 2,086EUROPE 1,681 1,406 1,608Spain 608 597 636Italy 685 436 572Greece 355 330 370Portugal 26 37 25France 2 1 2Albania 3 1 1AFRICA 153 180 299Tunisia 75 121 204Morocco 53 41 53Libya 10 10 8ASIA 159 236 164Syrian 43 102 71Turkey 96 97 58SOUTH AMERICA 10 10 12Argentina 9 9 10Source: FAO (1994) "Production Annual."

At the present time Peruvian olive groves in production are in:

1. Dpto. Lima: Huaura, Huaral, San Isidro, Lurín, Chilca, Bad, Cañete.

2. Dpto. Ica: Pisco, Paracas, Villacurí, be Born.

3. Dpto. Arequipa: Bella Unión, Acarí, Yauca, Jaqui, Cháparra, Mochica, Atiquipa, Chalaviejo,

Atico, Ocoña, Camaná, Mollendo, The Ensenada, Tambo, Cocachacra.

4. Dpto. Moquegua: Lomas de Ilo, Moquegua, Ilo.

5. Dpto. Tacna: Magollo, La Esperanza, La Yarada, Los Palos.

In the Peru there are more than 6,000 hectares of olive groves in production. There are

some 2,000 hectares more than olive trees in development. 90% of the olive groves in the Peru

are the variety "Sevillana" and "Creole". The productivity in the Peru is approximately, 42

kg/tree/year (3).

The area of olive groves planted in the world and the varieties are shown in the following

table (Table 2):

6

Table 2: AREA OF OLIVE GROVES IN THE WORLD BY COUNTRY AND MOREFREQUENT VARIETIES

Country Area VarietiesSpain 2,200,000 PICUAL, HOJIBLANCA,

Ecijano, Cornicabra, Arbequina, PicudoItaly 2,000,000 FRANTOIO, LECINO

Moraiolo, Cratina, Ascolana, Carolea Portugal 600,000 GALEA

Verdal, Redondil, Carrasquenha, Negrinha Greece 550,000 KORONEIKI

Conservolea, Kalamata, Amigdaloia France 40,000 PICHOLINE

Salonenque, Lucques, Aglandeau, Verdale Turkey 700,000 EDREMIT

Ayvalik, Domat, Izmir, Cakir Syrian 400,000 SOURYLebanon 30,000 BELADIJordan 55,000 CHAMIIsrael 10,000 MANZANILLAEgypt 2,000 WARDAMLibya 100,000 ENDURU, RASLITunisia 700,000 CHEMLALI, CHETOURI, MESKI, OUSLATIAlgeria 200,000 CHEMLAL K., SOGOISMorocco 350,000 PICHOLINE M.Cyprus 15,000Albania 40,000Yugoslavia 5,000California 150,000 MANZANILLO, GORDAL, MISSIONMexico 15,000 NEVADILLO, MISSIONPeru 7,000 ARAUCOChile 4,000 AZAPAArgentina 84,000 CRIOLLA

ArbequinaBrazil 800Uruguay 900Iraq 10,000Afghanistan 13,000China 7,000Angola 400South Africa 2,500Australia 2,000Japan 200

Source: Guerrero (1997) La nueva olivicultura

7

MORPHOLOGY OF THE OLIVE TREE

2.2.1 GENERAL CHARACTERS

The great longevity of the olive tree is a known fact. Cultivated trees exist, even in good

state of production, 300 to 400 years and can live up to 1000 years (19, 25). Also, should the

trunk die due to aging, when cut level with earth, it can sprout again from its base, giving rise to

a new tree. In fact, there are plantations that, due to aging of the trunk or because they died by

accident, such as strong icing, have been cut at the base and regenerated, having trunks much

younger than their root systems (19, 25).

The olive tree is a very rustic plant. It is found in lands of little fertility and in extremely

arid climates. When it is planted in fertile lands or places where there is good rainfall, production

is much higher (19).

The development of the tree varies according to the variety and media in which it is

grown. It sometimes reaches great size, although cultivation techniques limit the height of the

tree to make their exploitation easier (19, 40). The aerial portion is recognized by its dense

shape, short internodes and compact nature of the foliage (25).

2.2.2 ROOT SYSTEM

Root development of an olive tree depends greatly on the texture of the soil. In sandy,

loose soils, they develop deeper than in loamy, compact soil (19). The root system is shallow,

being in the top 0.6-1.2m in deep soils, with 70% of the roots in the first 0.6m (14, 19, 25).

Lateral roots extend out considerably, intertwining with those of adjacent olive trees over a very

wide area of a plantation (19).

2.2.3 LEAVES

The leaves are simple, whole, with a short petiole, growing parallel to the limb, generally

lance-like, and thick (19, 25). Each leaf lives approximately 2 years and falls in the spring, but as

with other evergreens, leaves can be on the tree for more than 2 years. Yellow colored leaves that

are still on the tree in the spring are indicative of either an abscission process a disease problem

or a nutrition problem (19, 25). The stoma is surrounded by a waxy skin that restricts the loss of

water, which makes it drought resistant.

8

2.2.4 FLOWERS

The inflorescence is a cluster that is born in the curvature of each leaf. Each cluster has a

variable number of flowers, which is dependent on the variety and the yearly stage of

development. It can vary between 10-40 flowers per cluster (19, 25).

The bud is usually formed in the area of previous growth and begins to be visible in the

area of new growth. The buds can remain dormant for more than a year before beginning to grow

forming viable inflorescence (25).

The flowers consist of four sepals, four petals, two stamens and two carpels. The bowl is

gamophyllous and the corolla gamopetalous. The stamens are inserted in the corolla. The carpels

are welded in a bilocular ovary. The stylus is generally short and bífid (19)

At each location two types of flowers are present: perfect flowers (with stamens and

pistil) and staminate flowers (with sterile pistils and functional stamens) in a proportion that

varies according to the variety, the inflorescence and the year (25).

2.2.5 FRUIT

The fruit is a drupe. The exocarpe or skin is free of hair has stomas and is joined to the

mesocarp, which is the pulp of the olive, and the endocarp that forms the stone that protects the

drop (19, 25).

2.3 YEARLY VEGETATIVE CYCLE

The olive tree begins its vegetative cycle at the beginning of the spring (September-

October), when new bud terminals are observed and with the appearance of the axially buds.

Flowering takes place in November-December and once the pollination is carried out fruit

development starts. In January-February hardening of the stone takes place. From this point the

fruit puts on weight until reaching normal size in April. Maturation starts in April. The duration

of this period depends on the variety (19).

The physiologic process that guides the flowering in the spring begins the previous

summer. The vegetative buds present in the curve of each leaf change becoming vegetative buds

or inflorescence and as a result contains flowers. In the summer the environmental factors

interact with the physiology of the tree to begin the induction process. The induction occurs

through chemical changes in the vegetative buds that cause the conversion to floral buds (27).

9

The process of floral development depends on good nutrition, for example, excess in

nitrogen can increase the quantity of flowers in some cases and in others it can cause them to

diminish (27). The process also depends on the temperature. According to Hartmann (1953) and

mentioned by Guerrero (19) the flowering and fruiting have a relationship with the number of

cold hours the olive tree is subjected to. It is always good for the flowering to coincide with days

of moderate temperature and when relative humidity is not too low. Martin (27) mentions that

the occasional occurrence of heat and dry winds during flowering can diminish the quantity of

fruit.

2.4 NEMATODES AND THE PLANTS

It has been often mentioned that the damages caused by nematodes will always be

proportionate to the population level of harmful varieties. In normal agricultural soil there is as

minimum of between 2,000 and 5,000 nematodes per each 100 cc of soil. Of them the biggest

percentage are saprophytes, followed by the phytophagous leaving a few predators (16).

Experimental studies, in general, indicate that the weight of the plant is inversely

proportional to the number of pathogen nematode present in the soil around the roots of the

plants. This relationship varies according to the plant and class of nematode, and it is subject to

the influence of environmental factors such as fertility, humidity, temperature and soil type. If

plants have an appropriate supply of food and appropriate environment, nematodes as any other

organism decrease logarithmically. The perennial plants provide a constant supply of foods;

therefore they are more vulnerable to the damage caused by these organisms (29). It is for this

reason that the results of the investigations in the control of nematodes is highly valid in the

place or area where this study was undertaken (7, 30).

2.4.1 EFFECTS OF M. incognito ON THE OLIVE TREE

It is well known that growth and reproduction of obligate parasites is associated with

their habitat (31), creating a very specific relationship between the nematode species or race with

the plant species or variety (32). The types of plant habitats differ in their capacity to allow the

reproduction of nematodes. These two characteristics can be independent (32).

As pathogens, nematodes affect yield or quality or both. They limit the use of the

nutrients for the plants, causing a waste of fertilizers. They increase perennial plants potential to

10

winter damage. Plants infected by them, wither quicker than those not infected. Certain species

act as vectors of pathogenic viruses, others alter the physiology of their host, in such a way that

they become more susceptible to fungal disease, or they provide the conduits for the entrance of

bacterial pathogen (29).

Quispe, in an experimental work carried out in 1992 (34), mentions that the olive tree is a

plant highly susceptible to attack by root knot nematode that cause decreased root development.

Root knots make in the absorption of nutritious inefficient and consequently a decrease in the

yield and quality of fruit. McKenry (28), also mentions that highly infested plants cannot usually

complete their normal functions, have reduced vigor and show symptoms decline. Also, the

incidence and damage caused to the olive tree depend on the texture of the soil and of the parent

variety. Fraga (16) also mentions that the olive tree is an efficient host for different species of

root knot nematode, which is, consequently, a limiting factor in the cultivation of this fruit-

bearing tree.

Robles (37) also mentions in Ica that the olive tree is one of the cultivations that is

affected by the root knot nematode and that the plants are attacked in inverse relationship to its

root development and in direct relationship to its permanency in the soil.

2.5. CHARACTERISTIC OF ROOT KNOT NEMATODE

The gender Meloidogyne, Goeldi, 1887, commonly known as root knot nematode, is

considered among the five pathogens that most affect the quality and quantity of foods of the

world (12, 22). The plants harmed by these nematodes are literally countless, however there are

some where the nematode action frequently becomes a restrictive factor for its cultivation (22,

38). Some of these are tomato, soy, grapevine, carnation, olive tree, cotton, cucurbits as a general

classification, etc. (16).

Meloidogyne is characterized by the marked sexual dimorphism of the adults, although

the male has the long and cylindrical characteristic form of most nematodes, the female swells

considerably and displays an augmented form, like a pear (12, 46). The males are 1.5 mm long

and the female 0.8 mm long. (16).

11

The reproduction is generally parthenogeneic and sometimes amphimictic. The female

after the fecundation deposits her eggs (200-500 up to 1000) in a mucilage mass that protects

them. The juveniles are born inside an egg; there they undergo their first change, after which

they soon leave to the exterior in search of a rootlet in which to settle. Once emergence occurs,

the juveniles need to find a rootlet quickly, since if they do not, they will die in few hours (at 12

hours 90% die and at 19 hours 99%). The juveniles that overcome this difficult stage and find an

available rootlet begin to feed immediately at the first point with which entry is made from the

exterior in order to recover from the effort required to carry out the function. Then, with their

stylet they punch a hole from which finally they can be introduced. Located in their definitive

position, they continue feeding voraciously. These juvenile, then go through two more stages

while continuing to feed. At the end of the third and onset of the fourth stage, the females begin

to adopt their pear shape form. The males, on the other hand, stay philiform through the third and

fourth stages, after the fourth change the adults leave to look for females. Once the females reach

their mature stage they can remain in the same place in which they were developed or, ripping

the semi-decomposed tissue, move to the root wall and stick out its exterior (16).

The cycle of life and the reproduction index vary with effects due to several factors, which

are important in determining the potential of reproduction of a species (31). For Taylor and

Sasser (42), temperature, longevity and humidity mainly determine the duration of the life cycle.

According to Fraga (16), the complete cycle of the species of Meloidogyne is completed in

approximately 30 to 45 days and in warm places or hothouses there can be up to twelve

generations annually. Temperature plays an important role in the development of root knot

nematode. Tyler, as mentioned by Christie, (13), observed that at temperatures of 27.5°C to 30°C

females developed from juvenile to fertile females in about 17 days. At 24°C the same change

takes place in 21 to 30 days; and at 20°C in 31 days and at 15.4°C in 57 days. At temperatures

lower than 15.4°C or higher than 33,5°C, the females don't reach maturity. Warm climates with

light and humid soils favor the development and the diffusion of this species, but an excess of

moisture is harmful (16, 33).

Longevity has not been fully studied among the species of Meloidogyne, some studies

indicate that the females can produce eggs for 2 to 3 months and are able to live a short time

afterward. (42). Nematodes display mechanisms to prolong their longevity during unfavorable

12

conditions (44), among the mechanisms the following can be mentioned: criobiosis, criptobiosis,

anhidrobiosis and anaxobiosis (31).

High population density can affect the development of nematodes as well as the relationship

between sexes (31). Fraga (16), mentions that a juvenile can become either genotype, they will

become a female if food is sufficient and a male if it is scare.

2.6 CONTROL OF NEMATODES

At the present time there are several effective methods to control nematodes, although

certain factors, such as the costs and cultivation practices limit their applicability in certain cases

(1). Four general types of control methods are used: cultural, physical, chemical and biological

(44).

2.6.1 METHODS OF CONTROL FOR ROOT KNOT NEMATODE

All the above methods have been proven possible to combat these nematodes, although it

has been verified that total eradication is difficult. However, good effects in crops can be

achieved by applying diverse measures (46).

Several studies have been carried out on the control of M. incognito with chemical

products and very recently, due to the toxicity of the chemical products and the unbalance

believe to be caused in the environment, with biological products.

In 1965, Furney (17) demonstrated in North Carolina that Temik is a promising pesticide

for the control of nematodes in tobacco when applied in proportions of 13.75 ú 8.625 kg/ha.

Nematodes present were Meloidogyne Goeldi and Pratylenchus Filipjev. Temik compared

favorably with DD and Viden D.

Martin and Birchfield (26), proved the effectiveness of DD, Vortex and SD-14647 Code

1-2-1-9 injected into the soil, and Temik and Mocap, applied as a granular to yam var.

Centennial. All the treatments reduced the population of the nematode. Temik and Mocap (4 kg

i.a./ha) maintained nematode population at a low level for up to 118 days after application. These

products showed 1 to 2 females per yam. Temik had the biggest yam yield.

Dickerson, Roblins and Greig (15), carried out 2 experiments on yam variety Tehoma,

planted in polyethylene bags and later transplanted into a definitive field,. In 1966 they proved

13

that Zinophos 10G and Temik 5G (1.04 g/30 cm2) increased production nearly 40%. However,

Zinophos slowed the emergence and development of the nursery plants, however, they recovered

in the field. In 1968, Mocap 10G, Disyston 10G, Temik 10G and Lannate 5G were tested. The

production of the treatments, when compared to control increased by 38% with Mocap, 23%

with Disyston, 19% with Temik and 16% with Lannate. The number of juveniles recovered, in

relation to the control, was lower for all treatments. Mocap impeded the growth of seedlings but

they recovered in the field and behaved similar to the control.

Angeles (2), evaluating a field of Japonica variety yams treated with Lannate 90W (9

kg/ha), Dupont 1410 (13.4 kg/ha), Temik 10G (8.4 kg/ha), Nemacur P (21 kg/ha), Mocap P (8

kg/ha), for control of M. incognito, demonstrated that Temik and Mocap were the most effective

treatments for the internal infestation of the tuber. Dupont 1410 was less effective in protecting

tubers from the infection of M. incognito.

In 1970, Tarjan et al (41) treated 12 year-old cocoa trees infested by Helicotylenchus,

Pratylenchus, Tylenchus, Meloidogyne, Trichodorus Cobb and Xiphinema Cobb with DBCP

(dibromodoropropano) applied, in water emulsion at 70 pounds per acre, Terracur (Dasamit)

granulated and granulated Mocap at 30 pounds per acre each, and granulated Nemacur at 20

pounds per acre. The harvested fruit of each of the experimental trees during 9 months were

counted and weighed. Comparing the number and weight of fruit, the treatment with Nemacur

exceeded the control respectively by 192% and 131%. The treatment with Terracur exceeded the

control by 142% and 103%, while the treatment with Mocap exceeded the control by 179% and

104%. Although the trees treated with DBCP produced 13% more fruit than control, their weight

was slightly smaller, which suggested possible phytotoxicity at the application rate.

Cabanillas and Chinchay (8), carried out a test in a field of tomato severely and evenly

infested with Meloidogyne. spp with four nematicides at two different rates: Furadan 5G

(carbofuran) at 50 and 100 kg/ha, Miral 5G at 25 and 50 kg/ha, Terracur P 10% (fensulfothion)

at 35 and 70 kg/ha, Vydate L 24% (Oxamyl at 2 and 4 l/ha). Two applications were made, the

first seven days after transplant and a second 45 days after the first application. All the

treatments were better than the control in regard to yield. Miral 5G at the different rates had the

highest yield the smallest yield was obtained with the treatments of Vydate. Also, the treatments

14

with Miral 5G were more effective in the reduction of the populations and nodulation index

caused by Meloidogyne spp.

Quispe (34) carried out a comparison of nematicides on olive trees for control of M.

incognito. Temik 15G at a rate of 150 g/plant, Mocap 10G at 300 g/plant, Curater 5G at 300

g/plant, Nemacur at 450 g/plant and HUNTER at 50 ml/plant. Temik 15G, Curater 5G and

Nemacur 5G reduced the population of M. incognito drastically, lasting 90 days after application

until initial levels were again reached. In the case of HUNTER (biological nematicide), the

population's reduction was slow but lasted 50 days, also, it didn't reduce the levels as low as with

the other products. The control maintained the initial population level in all of the evaluations.

The olive production was very reduced, mainly due to other factors, which is the reason for not

taking into consideration the effect of these products on yield during this study.

Udalova (43) mentions that some secondary metabolites of the plants, in particular the

alkaloids, are one of the factors that determine the resistance of plants to pests. He tested

amaranthina preparations (alkaloid taken place by Amaranthus cruentus) on tomato against M.

incognito. The tomato plants, susceptible to M. incognito, were sprayed with 50 and 100 ppm of

the solution of amaranthina before infestation. Analyses of isolations of nematodes of treated and

not treated plants showed a decrease in the number of nematodes per gram of root (2.1 times at

50 ppm and 1.3 times at 100 ppm in comparison to the control). The volume of females in the

treated plants was significantly smaller and it also decreased the fertility of the females (15-20%

in comparison with the control). The average weight of the treated plants was 30 % larger than

that of the control plants.

Lara et al (24) evaluated the effectiveness and profitability of the fungus Paecilomyces

lilacinus as a biological control for nodular nematode in tomato. P. lilacinus reduced the

populations of M. incognito in the soil and in the roots, parasite eggs of the nematode, reduced

root nodulation and increased yield and economic benefits for the cultivation. This organism

didn't affect the populations of the nematode Rotylenchulus reniformis or Helicotylenchus

dihistera.

Guevara (20) carried out experiments on how P. lilacinus was effected by some

nematicides commonly used in the control of M. incognito in olive tree. The following

treatments were used: Aldicarb (3 kg i.a./ha), Carbofuran (1,5 kg i.a./ha), Fenamiphos (1,5 kg

15

i.a./ha), Oxamil (3 kg i.a./ha) and P. lilacinus (1 kg of rice/infected tree). Results show the

variable action of the nematicides in the control of root knot nematode, related to the period of

action or effective residual power. With the application of P. lilacinus there was uniformity in

the control of the nematode. Where 30-90% of eggs and juvenile were infected with this

organism, the population of nematodes declined, this had an action different from that of the

nematicides which maintained the nematodes population level, avoiding in many cases the

problem of overcoming initial population levels.

Another type of control method for root knot nematode is physical control. In this control

type heat treatments can be used. Elevating soil temperature with steam or hot water to 50°C for

30 min. is enough to kill most nematodes and there eggs (1).

Some cultural control measures for M. incognito are deep, harrowed plowing,

scarification, etc., exposing the nematodes to the sun, to drying, to the wind, separating them of

the plant host, or mechanical damage (44).

Organic fertilizer used to improve soil texture conditions and fertility also decreases the

presence of nematodes and increases yields (33, 46). Works performed in Peru on cotton highly

infested by Meloidogyne, demonstrated that the addition of manure, especially when

incorporated into the soil of Crotalaria, like fertilizer, improved crops (46).

Presently, the only really positive method of control of Meloidogyne spp. is crop rotation

with resistant grass crops for at least two years (16, 33, 46). Tagetes patula is one of the

cultivations that are effective in decreasing populations of root knot nematode (33).

For fallowing to be effective, it needs to last two years and must be absolute, since any

overgrowth that develops during this period will be a perfect refuge for Meloidogyne spp. until

the next cultivation (16, 33).

There are no effective control methods of root knot nematode infestations in established

olive tree plantations. Fumigation of the soil prior to transplant and the use of stock free of

nematodes are the best methods to avoid a future damage in young trees (28).

The University of California has developed and patented the olive clone stock, Allegra,

which shows resistance to Meloidogyne spp. and to Verticillium spp. in laboratory tests (28).

16

III. MATERIALS AND METHODS

3.1 GEOGRAPHICAL LOCATION

The experimental field was located in field 8 of the Alamein Orchard, on property of

Engineer Ricardo Letts Colmenares, situated in the Pisco Province, in the Ica district.

3.2 CHARACTERISTICS OF THE TEST AREA

3.2.1 TEMPERATURE

The temperatures recorded during the experiment are shown in the Table 3.

TABLE 3: MONTHLY TEMPERATURES DURING THE TEST (C°)

Temperature Sep98

Oct98

Nov98

Dec98

Jan99

Feb99

Mar99

Apr99

May99

Jun99

Maximum 24 26 27 28 30 31 30 30 27 24Minimum 14 16 18 21 20 22 21 20 16 14Average 19 21 22 24 25 26 26 25 22 19

3.2.2 SOIL

The soil in the experimental area is a sandy texture with a pH of 8.7 with the following

construction:

% sand = 85% % clay = 10%

% lime = 5% % organic matter = 0.7%

3.2.3 WATER

The water used for irrigation came from a well. Watering was done via a gravity flow

method. The water running through the channels is diverted by means of PVC siphons and flows

to tree wells by means of PVC tubes. The wells are on both sides of the trees. Irrigation was done

with a quantity of 2 cubic meters of water per tree.

3.3 EXPERIMENTAL DESIGN

The experimental used a Block Design Completely Randomized, with the following

characteristics:

17

Number of blocks : 4

Number of treatments : 5

Trees/treatment : 7

Distancing between trees : 10 m x 10 m

The treatments used in the experiment are shown in the following table (Table 4):

Table 4: TREATMENTS AND RATES OF APPLICATION

TREATMENT PRODUCT RATE OF APPLICATION T1 HUNTER 15 ml/plant*T2 BIO-BAC 20 ml/plant*T3 BIO-BAC+HUNTER 20 ML BIO-BAC + 15 ML HUNTER/PLANT*T4 ALDICARB (TEMIK 15G) 200 g/plantT5 Control Nothing Applied

* Each rate was diluted in approximately 5 liters of water and applied with a spray of water followed by irrigation.

3.3.1 CARACTERISTÍCAS OF THE EXPERIMENTAL AREA

The experimental area consists of trees 40 years of age of the variety Sevillana grafted on

stock of the Frantoio and Lechino varieties (mingled) at a spacing of 10m x 10m.

The experimental area was divided into 4 blocks where each block represented a replicate

consisting of 5 treatments distributed arbitrarily, as shown in the graph in Annex 1.

Each treatment consists of a line. Each line had approximately 11 trees with a uniform

crown, it is for this reason that not all the treatments are equally distanced. The distance between

plants is 10 m and the distance between treatments is 10 m. However, between treatment III-3

and III-4 there is a distance of 40 m, between treatment III-4 and IV-3 there is a distance of 20

m. Between treatment IV-4 and IV-5 there is 100 m and between treatment IV-2 and IV-1 there

is 50 m. 7 plants per treatment (line) were marked at random, 4 of which were used for the

mentioned samplings. The remaining ones were for improvisation, the designated plants were

marked with a cross (X), as shown in the graph in Annex 2.

3.4 MATERIALS AND EQUIPMENT

3.4.1 VEGETATIVE MATERIAL

18

The experimental area consisted of olive trees of the variety Sevillana (which is the

variety most planted in the South Cost of Peru) infested by root knot nematode and planted on

stock of the varieties Frantoio and Lechino (mingled).

3.4.1.1 CHARACTERISTIC OF THE PLANT

The experimental field consists of planted trees of the variety Sevillana. This variety is

characterized to be a tree of very erect growth behavior (8-11 m) and biannual. Their leaves are

long and wide, with a brilliant dark green sheaf and a gray green back. The fruit is large (13.5 g)

and slightly asymmetric with rounded apex. It is enjoyed as a table olive due to the large size but

it is not of good quality. It is used in salads. It is a tree resistant to the cold and needs a certain

number of cold hours to flourish, in warm years it has little production. It has a very bad root and

is usually grafted onto the varieties Lechino or Rapasayo. Besides the normal fruit it has other

parthenocarpy that stops development and matures early (19). It is susceptible to Pseudomonas

savastanoi, but resistant to Spilocea oleaginea. It is difficult to root needing stakes, it is for this

reason that transplanting is done (40).

3.4.1.2 CHARACTERISTIC OF THE ROOT STOCK

The variety Sevillana is grafted onto rootstocks of Lechino and Frantoio, which are

mingled throughout the entire experimental field.

- LECHINO

This is a vigorous rustic variety. It is biannual, sensitive to ice and droughts resistant. It is

sensitive to disease caused by P. savastanoi (it produces tumors that initially are small, soft, flat

and green that later lignify, harden and have an irregular and cracked surface). The leaf has the

brilliant green sheaf and gray green back and is relatively small. The fruit is medium to small,

with rounded apex and stuck well to the tree. The yield of oil is medium to good and is of

considerable quality. It is a variety considered an excellent root stock for Sevillana, which is

grafted on it and produces fruits of better quality, and shape (bigger proportion of hardened fruit)

and size (19).

19

- FRANTOIO

This is a small rounded tree of medium vigor, with horizontal branches. The leaves are

lance like, with a green sheaf and silver back. It is a self-fertilizing variety. Fruit has a stepped

maturity; it yields very high oil with good quality. It is sensitive to Cycloconium. Although it is

quite demanding, it seems to adapt with ease to other media (19).

3.4.2 EXPERIMENTAL MATERIAL

The nematicide treatments that were used are described below:

- HUNTER

This is a preparation of plant extracts form Opuntia, Rhus, Rhizophoria, Quercus, and

other plants. It has desert mineral extracts, fatty acids and activated water (4, 36, 39). It is a

liquid, with 130 mg/l organic material and solids totaling 0.6%. Analysis show nucleic acid,

cytokinin, glycosides, porfrins, vitamin A, morphogenic substances and fatty acids that act as a

biocontrol that suppresses the development of nematode eggs and juvenile development (4, 36,

39).

The biocide activity of HUNTER in the root area it is based on the hydrolysis of

glycosides and phenolic substances that kill parasitic nematode and inhibit the development of

pathogenic fungi (27). HUNTER uses a repellent action against nematodes through the formation

of phenolic substances undesirable to the parasitic nematode (personal conversation with the

Engineer Ravines). Also, the nucleic acid derivatives activate cianobacterias by liberating

ethylene and hydrogen sulfide, which are also toxic to nematodes. Also, it supplies an energy

source for the multiplication of fungi nematofages and predators of nematodes such as mites

insects and predatory nematodes (36). The organic compounds in HUNTER diminish the content

of free amino acids in treated plant. This condition decreases the nematode population (4). Also,

these components increase photosynthesis, increasing energy capture, carbon fixation and

mineral uptake by the plant, when this occurs the general, health of the plant improves which

improves resistance to nematodes and other pathogens (4).

This multi-compound product is beneficial in the secondary control of invaders like

Pythium and Phytophthora that are present in fields. For this reason it can provide a better

control in field situations where these facultative pathogens are present (39).

20

- BIO-BAC

This is a nematicide that contains the following active ingredients: Bacillus subtillis (2

strains), B. cereus, B. megatarium, B. thuringiensis, Azotobacteria vinelandii, Azotobacteriaceae,

mocrococcus, species of Pseudomonas, Rhizobium japonicum, R. leguminosarum, extract of

Aspergillus orizae, cultures of Lactobacillus with nutrients, D. brochapaga, and A. oligospora.

It also contains A. botriospora, an imperfect fungi that captures nematode, and micronutrients

that support the growth and development of the plants (including copper sulfate, and zinc sulfate)

(23).

It helps in the promotion and development of antagonists increasing the activity of soil

microorganisms. It aids in the reduction of soil compaction. It adds vital trace elements. It is

good as a seed inoculant. Beneficial bacteria are inoculated into sterilized soil w/o alkaloids. It

controls nematodes biologically with trapping fungi. (23)

BIO-BAC uses a natural biological mechanism to control soil borne nematodes. BIO-

BAC contains species of fungi that are parasitic. Because of this, they destroy nematodes and use

them as a nutrition source. More precise, imperfect fungi trap nematodes that inhabit rich soils.

Using bundles and sticky loops they catch nematodes in the soil. Once caught, the fungi invade

the nematode dissolve and consume it. The trapping of the nematode is made with the use of a

compound called "lectins". These compounds have a magnetic attraction to the nematodes

surface, the fungi then use their rings and bundles (23).

Another mechanism used for ensnaring is a small lasso that entangles the cells. These

lasso like structures, when readied for contact, expand then press the nematode catching it in the

loop. Once captured, the nematode is invaded by the mycelium of the fungi and digested

internally (23).

- ALDICARB (TEMIK 15 G)

This is a carbamate that has a contact and systemic action, whose chemical formula is

shown below:

21

CH3 O|| H

CH3SCCH=NOCNCH3

CH3

C7H14N2O2S

It has a molecular weight of 190.3 and the chemical name is 2-metyl-2-(metyltio)

propionaldehido 0-(metilcarbamoil) oxyme (45). It has an oral LD 50 of 7 mg/kg and a dermal

LD 50 of 2100-3970 mg/kg in rats (20). It is in a crystalline solid state with a white color. It has

a lightly sulfurous scent. It has a fusion point of 98°-100°C and a boiling point of 100°C (45).

It controls ecto and endo-parasite juvenile and adult nematodes, in annual and perennial

cultivations. The control takes place via contact when the nematodes are in free form in the soil.

They are controlled through systemic action when the nematodes feed enough on the tissue of

the plants to absorb it also getting the Aldicarb. Other forms of control are through repelling the

nematode from the roots treated with Aldicarb and through interference in the reproduction and

disorientation of the males. The eggs and cysts states are less susceptible to Aldicarb (45).

Aldicarb should not be applied to the foliage of the plants due to phytotixic problems.

The residual effect varies from 30-60 days depending on temperature, the higher the

temperatures the less the residual effect will be. Neither the type of soil nor pH affects its

systemic activity (20).

3.4.3 EQUIPMENT

3.4.3.1 FIELD MATERIAL

For samplings the following materials were used:

- Shovel - Labels

- Plastic bags - Buckets

For application of the treatments the following materials were used:

- Shovel - Watering-cans

- PVC Tubing (for watering) - Graduated test tube

- PVC Siphons (for watering) - Scale

22

3.4.3.2 LABORITORY MATERIAL

For the different extraction methods and to determine nematode nodulation index the

following materials were used:

Tray Method (10):

- Graduate test tube - Sieve

- Tray - Stereoscope

- Toilet paper - Plastic Basins

- Pipettes - Erlenmeyer

Centrifugation Method (11):

- Centrifuges - Plastic Basins

- Graduated test tube - Pipette

- Sieve - Erlenmeyer

- Sugar - Stereoscope

Root Dying Method (18):

- Precision balance - Pipette

- Sodium Hypochlorite Solution - Erlenmeyer

- Fuscina Lactofenol Acid - Stereoscope

- Plastic Basin - Microwave Oven

- Stirrer - Sieve

Bioassay (5):

- Gavel - Tomato Plants

3.4.4 FIELD WORK

3.4.4.1 Watering

Watering was carried out approximately every 25 days with 2 cubic meters of water per

tree. Before watering, the tree wells were skirted with a tractor and tubing was situated to draw

water into the well. Then, by means of siphoning, the water was diverted into the main channel

corresponding to each tree.

23

3.4.4.2 Fertilization

No application of fertilizer organic matter or gypsum was made to the soil, however,

foliar application with Polical, Nutribor, Magnisal, Kelatex Cu, Fetrilom Combi and urea was

made in September 98.

3.4.4.3 Phytosanitary Control

During the experimental work Palpita quadrastigmalis was detected and controlled by

means of light traps that were not only effective in trapping the adult of this species but also for

the adults of other species. Strips of paper were also used to capture the larva, which goes down

to the soil to pupate. The biological insecticide MVP containing indotoxins produced by Bacillus

thuringiensis also was applied for the control of lepidoptera. There were seven releases of the

micro-parasite Trichogramma for the control of the larvae of this insect.

There were very low populations of Saissetia that, according to Barnet (6), is one of the

main diseases of olive trees.

3.4.5 METHODS AND PROCEDURES

3.4.5.1 SAMPLING

Samplings began at the first plant marked with a cross (X) in the line corresponding to

each treatment, sampling followed on marked plants in the line. Each sample was confirmed by 2

sub-samples taken under the canopy at a depth of 30 cm, until having an approximate total of 1

kg of soil for samples as mentioned by Quispe (35). An aggregate of roots was obtained with

each sample, especially those that showed any suspicious disease symptom. The extracted

samples were individually placed in plastic bags and identified for later nematalogical analysis.

A total of 20 samples (one sample per treatment with four repetitions on each date) were

collected. The dates of sampling are in the table in Attachment 3.

3.4.5.2 APPLICATION OF TREATMENTS

The application of the different treatments were made as follows:

T1: 15 ml of HUNTER per plant diluted in approximately 5 liters of water was applied

according to the chronological diagram shown in Attachment 3. The applications were

carried out with a watering can with the rate applied under the canopy with a spray of

water.

24

T2: 20 ml of BIO-BAC per plant was applied diluted in approximately 5 liters of water,

according to the chronological diagram shown in the Attachment 3. The applications

were carried out with a watering can with the rate applied under the canopy with a spray

of water.

T3: 15 ml of HUNTER was applied with 20 ml of BIO-BAC per plant diluted in

approximately 5 liters give water, according to the chronological diagram shown in the

Attachment 3. The applications were carried out with a watering can with the rate applied

under the canopy with a spray of water.

T4: A single application of 200g of ALDICARB (Temik 15G) per plant was made, according

to the chronological diagram shown in the Attachment 3. It was incorporated into the soil

under the canopy to a depth of 15 cm.

T5: No nematicide application was carried out.

3.5 VARIABLES ANALIZED IN THE EXPERIMENT

3.5.1 POPULATION OF NEMATODES IN THE SOIL

The methods of extraction of nematodes to obtaining infectious states of Meloidogyne,

other parasitic nematode as well as non-parasitic nematodes in 100 cc of soil, was: the tray

method (10, 16) and the centrifuge method in sugar (11).

3.5.2 POPULATION OF NEMATODES IN THE ROOTS

The roots were processed using the sodium hydochlorite method, where the number of

Meloidogyne nematodes in infected roots was obtained. The roots were processed by staining

roots in fuscina lactophenol acid (18) that also showed other nematodes in 5 g of roots.

3.5.3 NODULATION INDEX

Bioassay tests were carried out, Goodfrey (1934) Soil Sci. 38: 3-27, mentioned by Barker

and Nusbaum (5), using soil from the plots and one in which tomatoes were transplanted over a

period of 3 to 4 weeks in hothouse. The root samples were weighed and root nodulation index

determined according to the scale established by International Project of Meloidogyne (42) that

is shown in the Table 5.

25

Table 5: INDEX OF ROOT NODULACIÓN

Degree Quantity of nodules0……………………………..No nodules or egg masses.

1..............................................1-2 nodules or egg masses.

2.............................................3-10 nodules or egg masses.

3............................................11-30 nodules or egg masses.

4..........................................31-100 nodules or egg masses.

5...............................more than 100 nodules or egg masses.

3.5.4 YIELD AND QUALITY OF YIELD

3.5.4.1 NUMBER OF FRUIT PER TREE

The number of fruit was counted from uniform trees of each treatment, and then the

average per tree was obtained.

3.5.4.2 WEIGH OF FRUIT

The weight of fruit was obtained by weighing a sample of harvested fruit form each one

of the trees per treatment then dividing that by the number of fruit sampled then obtaining an

average of all the trees. The obtained value is expressed in grams.

3.5.4.3 YIELD PER TREE

The yield per tree was obtained by multiplying the number of fruit per tree by the weight

of the fruit per tree. The value obtained is expressed in grams.

IV. RESULTS AND DISCUSSIONS

The results discussed in the following sections are: analysis and interpretation of the

effect of the nematicides on the populations of M. incognito in soil and root; index of nodulation;

populations of parasitic nematodes and non-parasitic nematodes; yield; and the effect the

nematicides had on the populations of beneficial nematodes.

4.1 POPULATION OF M. incognito

4.1.1 POPULATION OF M. incognito IN THE SOIL

As mentioned by Calzada (9) carrying out the analysis of variance logarithm for number

of juveniles in 100cc of soil is done to achieve a lower coefficient of variation because the results

vary from very low values to several thousands, (Attachment 6).

26

Table 6: Number of juveniles of M. incognito in 100 cc of soil in olive trees treated with two

biological nematicides, Aldicarb and untreated in fields infested with the root knot

nematode (Orchard Alamein-Pisco-Ica)

Treatment 1st sample 2nd sample 3rd sample 4th sample 5th sampleHUNTER 42 b 125 b 46 a 805 a 165 aBIO-BAC 40 b 123 b 24 a 353 ab 210 aHUNTER+BIO-BAC 77 a 267 a 41 a 415 ab 124 aALDICARB 63 a 132 b 37 a 511 ab 495 aCONTROL 39 b 376 a 23 a 262 b 340 a

As can be seen in Table 6 and Figure 1, no tendencies exist either upward or downward

except in the fourth sample where the population is higher in almost all the treatments. The data

for the populations of nematode in the soil are very variable, as is that for the distribution of

nematodes in the soil, possibly due to influences such as soil climate, biological influences and

chemical influences on the populations.

In the initial population (first sampling) the treatments with ALDICARB and the mixture

of HUNTER+BIO-BAC had a higher population density (63 and 77 respectively) and they are

statistically similar but different from other treatments which have an advantage with lower

populations.

Figure 1: Number of juveniles of M. incognito in 100 cc of soil in olive trees treated with

two biological nematicides, Aldicarb and untreated in fields infested with root knot


27

1st Sample4th Sample

Hun

ter

Biob

ac

Hun

ter+

Bio-

Bac

Aldi

carb

Con

trol

0.00

200.00

400.00

600.00

800.00

1000.00

In the second sampling there was no nematicidal effect observed in any of the treatments,

an increase in population is observed in all the treatments due to the progressive increase in

temperature, encouraging higher nematode reproduction that prevented the nematicides from

working efficiently during this whole period. There were no significant differences between

HUNTER, BIO-BAC and ALDICARB, but all were statistically different from CONTROL and

HUNTER+BIO-BAC treatments. There was a smaller increase in nematode populations in the

HUNTER, BIO-BAC and ALDICARB treatments.

In the third sampling a decrease in the populations in all the treatments is observed all

being lower than the initial population, except for the HUNTER treatment that had a population

similar to the first sampling. In spite of not having significant differences, the lowest populations

were in the BIO-BAC and CONTROL treatments. This drastic decrease in the population density

was not caused by the action of the nematicides alone but possibly due also to a watering made

one day before the sampling, possibly causing the nematodes to be washed below the 30 cm

sampling depth resulting in them not being counted.

28

In the fourth sampling, all the treatments again had increases in M. incognito populations

due to high temperatures. The HUNTER treatment had the highest number of J2 in 100 cc of soil

(805) and the CONTROL treatment had the lowest number (262).

In the fifth sampling, in spite of not being statistical different, the CONTROL continued

to have an increase in the population of juvenile in spite of a temperature decrease; while in all

the other treatments, nematode populations decreased. The ALDICARB treatment maintained

almost the same population as the previous sampling. The biological nematicide treatments

higher populations declined, with the HUNTER+BIO-BAC treatment having the lowest number

of juveniles. This decline in the populations possibly is due to lower temperatures in the area as

well as the action of the nematicides on juvenile of M. incognito in the soil. The ranking of

decline was, in order, BIO-BAC, HUNTER+BIO-BAC, HUNTER, CONTROL then lastly

ALDICARB.

4.1.2 POPULATION OF M. incognito IN THE ROOT

As mentioned by Calzada (9) high and low values were taken out of the logarithm for the

purpose of diminishing the coefficient of variation because the results vary from very low values

to several thousands (Attachment 7).

Table 7 and Figure 2 show that, in general, populations in the CONTROL increase

throughout the experiment, this is also true with the ALDICARB treatment, however, a

population decrease is observed in the fifth sampling (final population). The biological

nematicides had fewer juvenile and eggs than the two previous treatments. The treatment with

BIO-BAC was the most effective in maintaining an almost constant population, followed by the

HUNTER+BIO-BAC treatment that also maintained an almost constant population although

higher than the treatment with BIO-BAC. In the HUNTER treatment, populations increase until

the fourth sampling where it leveled off and diminished in the final population.

Table 7: Number of juvenile and eggs of M. incognito in 5g of roots of olive trees treated

with two biological nematicides, Aldicarb and Untreated in fields infested with Root Knot

Nematode (Orchard Alamein-Pisco-Ica)

Treatment 1st sample 2nd sample 3rd sample 4th sample 5th sampleHUNTER 1007 b 2219 a 3735 a 5661 a 2900 bcBIO-BAC 2217 a 2776 a 1157 b 1147 b 1583 c

29

HUNTER+BIO-BAC 2536 a 1700 a 2820 a 2583 ab 2167 bcALDICARB 2744 a 2727 a 4093 a 6220 a 5583 abCONTROL 1165 b 2343 a 7292 a 5787 a 15500 a

In the first sampling (initial population) there are no significant differences among the

BIO-BAC, HUNTER+BIO-BAC and ALDICARB treatments. However, these treatments are

statistically different from the treatments with HUNTER and CONTROL, which had lower

populations than the previous treatments.

In the second sampling, in spite of not having significant differences, the CONTROL

continued to have increased populations of M. incognito in the root, the same was true with

HUNTER and in smaller proportion with the BIO-BAC. This was due to the increased

temperature not allowing an effective action of the nematicides. The treatments with

HUNTER+BIO-BAC and with ALDICARB reduced the population of M. incognito in the root,

with the first having a larger reduction due to the mode of action.

30

Figure 2: Number of juvenile and eggs of M. incognito in 5g of roots of olive trees treated

with two biological nematicides, Aldicarb and Untreated in fields infested with root knot


1st S

ampl

e

3rd

Sam

ple

5th

Sam

ple

Hun

ter

Bio-

Bac

Hun

ter+

Bio-

Bac

Aldi

carb

Con

trol

0.02000.04000.06000.08000.0

10000.012000.014000.016000.0

In the third sampling the CONTROL continued to have an increase in the populations of

M. incognito. The populations also increased in the treatments with HUNTER, HUNTER+BIO-

BAC and ALDICARB due to the progressive increase in temperature that reduces the

nematicidal effects of the products. Only the treatment with BIO-BAC reduced the number of

nematodes in 5g of roots compared to the initial population, with this treatment being statistically

different from the other treatments.

In the fourth sampling, the CONTROL population decreased to 5787, whereas, the

treatments with HUNTER and ALDICARB had population increases to 5661 and 6220

respectively, which were similar statistically. The treatments with BIO-BAC and

31

HUNTER+BIO-BAC had a smaller number of nematodes per 5g of root having a reduced

population compared to both the previous sampling and the initial population.

In the fifth sampling the populations of the Control changed, being statistically similar to

the treatment with ALDICARB but different from the other treatments. The populations in the

treatments with ALDICARB, HUNTER and HUNTER+BIO-BAC declined. They previously

had a population larger than the initial one, then in the third sampling they had a population

smaller than the initial. Statistically, the treatments with HUNTER and HUNTER+BIO-BAC

were the same, and similar to ALDICARB. In the BIO-BAC treatment the population increased

slightly, but stayed less than the initial population and was statistically different from the control

and similar to the other treatments.

4.1.3 NODULATION INDEX

The results of the nodulation index are shown in Table 8 and Figure 3. As shown, the best

treatment was HUNTER+BIO-BAC, followed by BIO-BAC, HUNTER, ALDICARB and

CONTROL. The products of biological origin were an effective control of nematodes as far as

reducing the degree of nodulation in the roots, not only in the tomato bioassay, but also in the

roots of the olive trees. The CONTROL had the maximum root damage index, which was at the

point where absorption of water and nutrients from the soil was restricted. As a consequence of

this damage, fruit production was drastically reduced compared to the treatments where there

was an application of any product.

The nodulation indexes were similar for all treatments in the initial population. Although

not significant, the plants treated with ALDICARB had the lowest root nodulation index and the

CONTROL had the largest compared to the other treatments.

In the second sampling an immediate effect is observed for the nematicides, since the

nodulation index diminishes. There were no significant differences between the biological

nematicides and ALDICARB, with the HUNTER+BIO-BAC treatment having the lowest index

of nodulation and the CONTROL having the highest compared to the initial population (5.0 to

4.3).

32

Table 8: Index of nodulation caused by M. incognito in roots of tomatoes in soil of olive

trees treated with two biological nematicides, Aldicarb and untreated fields infested with

the root knot nematode (Orchard Alamein-Pisco-Ica)

Treatment 1st sample 2nd sample 3rd sample 4th sample 5th sampleHUNTER 4.8 a 4.0 ab 3.8 b 4.3 b 2.3 bBIO-BAC 4.8 a 3.3 ab 2.8 c 3.3 c 2.0 bHUNTER+BIO-BAC 4.3 a 2.8 b 3.0 c 3.0 c 2.0 bALDICARB 4.0 a 3.8 ab 4.8 a 4.5 ab 3.5 aTESTIGO 5.0 a 4.3 a 4.8 a 5.0 a 4.0 a

In the roots evaluated in the third sampling, a decrease is observed in the index of

nodulation in the treatments with the biological nematicides, also, in the treatment with

HUNTER+BIO-BAC, a small increase occurs but not significantly different from the treatment

with BIO-BAC. The treatment with ALDICARB had an increase in the index of nodulation

compared to the initial population, indicating an end to its residual effect. The CONTROL

treatment, due to the increase in temperatures, shows an increase in root nodulation compared to

the second sampling. The index of nodulation for this sampling is low, not only due to the action

of the nematicides, but also possibly due to a watering made one day before sampling washing

the nematodes beyond the 30 cm. sample depth.

In the fourth sampling, the index of nodulation increased (compared to the third

sampling) in all the treatments except in the HUNTER+BIO-BAC treatment, where the index of

nodulation remained at 3.0. The CONTROL had the largest index of nodulation, with a

significant difference occurring between it and the other treatments, this high value in the index

of nodulation compared to the previous sampling possibly was due to the temperature, that

stayed at 26°C.

Figure 3: Nodulation index indicating M. incognito in tomato roots planted in soil of olive

trees treated with two biological nematicides, Aldicarb and untreated in fields infested with

root knot nematode (Orchard Alamein-Pisco-Ica)

33

1st S

ampl

e

3rd

Sam

ple

5th

Sam

ple Hu n te r B io - B a c Hu n te r + B io -

B a c

A ld ic a r b Co n tr o l

0

1

2

3

4

5

In the fifth sampling a decrease occurred in the root nodulation index in all the treatments

when compared with the initial population. Even the Control had an index of 4.0, this due to the

drop in temperature that occurred in the area ending up at 19°C, as well as the action of the

nematicides. There were no significant differences between the Control and Aldicarb, but there

was a significant difference between the two treatments with the biological nematicides, and the

Control that had an index of 4.0.

In general, with the biological nematicides the nodulation index has a tendency to

decline, whereas, with the Control and Aldicarb a defined tendency is not observed in the first

three samplings, however, there is a decrease in the nodulation index due to the temperature.

4.2 POPULATION OF PARACITIC AND NON-PARACITIC NEMATODES

4.2.1 PARACITIC NEMATODE POPULATION

In the soil

Table 9 and Figure 4 shows the populations of parasitic nematodes in the soil (including

M. incognito) which follows the same tendency of the population of J2 of M. incognito in 100 cc

of soil. The same logarithm is used to show the large variations and to avoid a high coefficient

of variation (Attachment 8).

Table 9: Number of parasitic nematodes in 100 cc of soil in olive trees treated with two

biological nematicides, Aldicarb and untreated in fields infested with root knot nematode

(Orchard Alamein-Pisco-Ica)

34

Treatment 1st sample 2nd sample 3rd sample 4th sample 5th sampleHUNTER 104 a 207 bc 118 a 873 a 223 aBIO-BAC 126 a 211 bc 64 a 424 b 236 aHUNTER-BIOBAC 134 a 346 ab 95 a 436 ab 263 aALDICARB 122 a 186 c 78 a 543 ab 532 aCONTROL 104 a 474 a 62 a 370 ab 388 a

As can be seen, neither an upward or downward trend exists. The data on the populations

of parasitic nematodes in the soil is very variable in that there is much irregularity in the

distribution of the nematodes in the soil, possibly due to the influence of many climatic factors,

such as soil, biological influences and chemical influences on the populations.

In the first sampling, the populations of parasitic nematodes, varying from 104 to 134

nematodes per 100 cc of soil, are similar statistically to the initial populations.

In the second sampling, all the populations increased. The treatments with HUNTER and

BIO-BAC are statistically the same and similar to the treatments with HUNTER+BIO-BAC and

ALDICARB. The CONTROL is statistically different from the ALDICARB treatment, but

similar to the other treatments. The treatments with HUNTER, BIO-BAC and ALDICARB had

slightly increased populations, but when compared to the initial population it is seen that they

have stayed the same. The treatments with the biological nematicide mixture and the control had

the highest populations. No nematicide effect is observed because of the high temperatures that

caused an increase in the number of nematodes which voided the effective action of these

products during this period.

In the third sampling, a decrease in the populations in all the treatments occurred, being

similar and even smaller than the initial population, due possibly to a watering made the day

before the sampling, causing some of the nematodes to be washed beyond the 30 cm. sample

depth. In spite of being similar statistically, the treatment with HUNTER has the highest

population (118 in 100 cc of soil) whereas, the lowest populations were in the Control with 62 in

100 cc of soil (which was very similar to the treatment with BIO-BAC with 64).

Figure 4: Number of parasitic nematodes in 100 cc of soil of olive trees treated with two

biological nematicides, Aldicarb and untreated in fields infested with nematodes (Orchard

Alamein-Pisco-Ica)

35

H u n te rB io -B a c

H + BA ld ic a rb

C o n tro l

0 .0 0

2 0 0 .0 0

4 0 0 .0 0

6 0 0 .0 0

8 0 0 .0 0

1 0 0 0 .0 0

In the fourth sampling a population increase again occurs, probably due to the high

temperatures that caused high reproduction of the nematodes. The treatments with

HUNTER+BIO-BAC, ALDICARB and the CONTROL are similar statistically having

populations of 436, 542 and 3370 in 100 cc. of soil respectively. The highest populations were in

the HUNTER treatment and the lowest with BIO-BAC.

In the final population (fifth sampling) a decrease in the nematode populations occurs in

all treatments except the CONTROL that had an increase in the population of parasitic

nematodes in the soil, indicating that it maintained its populations. In spite of not having

statistical differences, the treatments with HUNTER, BIO-BAC, HUNTER+BIO-BAC and

ALDICARB, in that order, have the lowest populations. This decrease is credited to the

nematicidal effect along with a low temperature.

The treatment that decreased parasitic nematodes in the soil the best was BIO-BAC,

followed by HUNTER+BIO-BAC, ALDICARB, and CONTROL. Had the HUNTER treatment

not had a spike in the fourth sampling it would also be one of the best treatments. This spike

took place due to the high temperatures registered in the area and also due to the lack of organic

matter, that in the case of HUNTER is very important (conversation with Engineer Ravines).

In the root

As shown in Table 10 and the following Figure, only in the treatments of BIO-BAC and

HUNTER+BIO-BAC did the populations of parasitic nematodes in the root (including M.

incognito) stay steady throughout the experiment. In other words, they were the best treatments

36

in the control of parasitic nematodes in the root. The other treatments (HUNTER, ALDICARB

and CONTROL) had population increases until the fourth sampling when it stopped then

diminished or leveled off. In the Control, the populations increased proportionally higher than

the other treatments until reaching a population of almost 10000 phytoparasites in 5g of soil. One

can affirm then that there was a nematicidal effect in the control of parasitic nematodes in the

root. As in previous areas the logarithm was carried out due to the high variables in the values

that were obtained in the results and to avoid a high coefficient of variation (Attachment 10).

In the initial population the treatments with HUNTER and the CONTROL are statistically

similar, in the other three treatments there are no statistical differences and they had slightly

higher populations, meaning they are starting at a disadvantage.

Table 10: Number of parasitic nematodes in 5g of root in olive trees treated with two


Alamein-Pisco-Ica)

Treatment 1st sample 2nd sample 3rd sample 4th sample 5th sampleHUNTER 1017 b 2227 a 3735 a 5661 a 2900 abcBIO-BAC 2247 a 2786 a 1157 b 1147 b 1646 cHUNTER+BIO-BAC 2554 a 1705 a 2832 a 2583 ab 2173 bcALDICARB 2774 a 2729 a 4093 a 6220 a 5708 abCONTROL 1183 b 2348 a 7292 a 5787 a 9806 a

In the second sampling statistical differences are not observed, however, only in the

treatments with HUNTER+BIO-BAC and ALDICARB did the populations diminished slightly.

The other treatments had increases in the populations of parasites in the root, also slight, making

it possible to affirm that the populations in all the treatments stayed similar to the initial

population in judging the effect of the nematicides during this period.

Figure 5: Number of parasitic nematodes in 5g of root in olive trees treated with two


Alamein-Pisco-Ica)

37

1st S

ampl

e

2nd

Sam

ple

3rd

Sam

ple

4th

Sam

ple

5th

Sam

ple

H u n te rB io -B a c

H + BA ld ic a r bC o n tr o l 0 .0 0

2 0 0 0 .0 0

4 0 0 0 .0 0

6 0 0 0 .0 0

8 0 0 0 .0 0

1 0 0 0 0 .0 0

In the third sampling, the CONTROL and the treatment with HUNTER continued to have

increased populations (the later more drastically than the former). The same affect happens to

HUNTER+BIO-BAC and ALDICARB treatments where the populations of parasitic nematodes

increase in the root, increasing higher than the initial population, this was due to an increase in

temperatures. In the treatment with BIO-BAC the populations decrease and it is statistically

different to the other treatments indicating a product action in spite of the high temperatures.

In the fourth sampling it is observed that the treatments with ALDICARB and HUNTER

are similar statistically and in these the populations increased compared to the previous

sampling, with the HUNTER treatment having a spike of 5601 nematodes in 5g of root. The

Control had a decrease in the populations having a similar value to the two prior mentioned

treatments. The BIO-BAC continued having a decrease in populations ending up being lower

than the initial sampling. With HUNTER+BIO-BAC a slight increase in the populations

occurred, but they ended up being similar to the initial population. The highest populations were

in the treatment with Aldicarb with 6220 nematodes in 5g of root and to a lesser extent the BIO-

BAC with 1147 nematodes in 5g of root.

In the fifth sampling a spike occurs in the populations in the control treatment being nine

times that of the initial population. The treatment with HUNTER had decreased populations

being similar to the second sampling. In the treatments with BIO-BAC, HUNTER+BIO-BAC

and ALDICARB the populations stayed similar to the previous population.

4.2.2 POPULATIONS OF NON-PARASITIC NEMATODES

38

In the soil

Figure 11 and Figure 6 show that there is a slight tendency in the increase of populations

of non-parasitic nematodes in the soil in all the treatments. More specifically, the Control

treatment maintains consistent populations throughout the experiment. In the other treatments the

populations have a slight increase in the second sampling, and a slight decrease to the third

sampling. In the fourth sampling the increase is bigger, it levels off then diminishes in the fifth

sampling ending up equaling the initial population. The treatment with the smallest number of

non-parasitic nematodes in the soil is the CONTROL, followed by BIO-BAC, HUNTER+BIO-

BAC, HUNTER and ALDICARB. As before the logarithm was used to take out the big

variations that exist in the results in order to avoid a high coefficient of variation (Attachment 9).

In the initial population (first sampling) all the treatments have similar populations, with

only small statistical differences.

In the second sampling all the populations are similar statistically, however, in the

treatment with HUNTER, BIO-BAC, ALDICARB and in the CONTROL the populations

increase due to an increase in temperature. That didn’t occur in the treatment with

HUNTER+BIO-BAC where the populations stay similar to the initial population, due to the

mode of action of the nematicide, which was not effected by the elevated temperatures.

Table 11: Number of non-parasitic nematodes in 100 cc of soil in olive trees treated with

two biological nematicides, Aldicarb and untreated in fields infested nematode (Orchard

Alamein-Pisco-Ica)

Treatment 1st Sample 2nd Sample 3rd Sample 4th Sample 5th SampleHUNTER 41 ab 96 a 77 a 190 a 84 aBIO-BAC 92 a 113 a 84 a 147 a 84 aHUNTER+BIO-BAC 71 ab 68 a 90 a 155 a 66 aALDICARB 59 ab 119 a 107 a 211 a 86 aCONTROL 29 b 74 a 76 a 92 a 74 a

In the third sampling the populations are also statistically similar with a slight decrease in

the treatment with HUNTER, BIO-BAC and ALDICARB and a slight increase in the treatments

with HUNTER+BIO-BAC and the CONTROL. The populations stay similar to the previous

population and are similar to the initial population. One could say that the nematicides didn't

allow the populations to increase but rather they stay constant throughout these three samplings.

39

Figures 6: Number of non-parasitic nematodes in 100 cc of soil in olive trees treated with

two biological nematicides, Aldicarb and untreated in fields infested nematodes (Orchard

Alamein-Pisco-Ica)

1st S

ampl

e

3rd

Sam

ple

5th

Sam

ple

H u n t e rB io - B a cH + BA ld ic a r bC o n t r o l0 .0 0

5 0 .0 0

1 0 0 .0 0

1 5 0 .0 0

2 0 0 .0 0

2 5 0 .0 0

In the fourth sampling, due to the increase in temperature, all the treatments increased the

populations dramatically except Control.

In the fifth sampling a decrease again occurs in the populations of all the treatments

becoming similar to the initial population due to the drastic decrease in temperatures.

A high number of non-parasitic nematodes in the soil would indicate a lesser effect of the

nematicide products in that beneficial nemato-fauna and a greater biodiversity could negatively

affect parasitic nematodes. A higher number of non-parasitic nematodes in the soil result in a

lower number of parasitic nematodes.

In the root

Table 12 and Figure 7 show the results of the populations of non-parasitic nematodes in

the root. Attachment 11 gives the analysis of variance where the results were subjected to the

logarithm that decreased the coefficient of variation.

The treatment with BIO-BAC is the only one to maintains similar populations throughout

the experiment, which affirms that it is the treatment with the most effect on the non-parasitic

nematodes in the root, followed by the treatment with HUNTER, HUNTER+BIO-BAC,

CONTROL and ALDICARB.

40

In the initial population the populations are similar. The Control has the lowest and the

treatment with HUNTER+BIO-BAC the highest population.

Picture 12: Number of non-parasitic nematodes in 5g of root of olive trees treated with two

biological nematicides, Aldicarb and untreated in fields infested nematodes (Orchard

Alamein-Pisco-Ica)

Treatment 1st Sample 2nd Sample 3rd Sample 4th Sample 5th SampleHUNTER 25 ab 30 a 55 ab 335 a 150 aBIO-BAC 15 bc 30 a 30 b 100 a 25 aHUNTER+BIO-BAC 50 a 10 a 57 ab 162 a 312 aALDICARB 45 a 10 a 15 b 170 a 525 aCONTROL 5 c 5 a 242 a 102 a 252 a

In the second sampling there were no statistical differences in the population of non-

parasitic nematodes in the root. However, the CONTROL treatment maintained the initial

population, whereas, the other treatments, in spite of having slight increases (HUNTER, BIO-

BAC) or slight decreases (HUNTER+BIO-BAC, ALDICARB) the populations didn't vary a lot

compared to the initial population, indicating the nematicides didn't largely affect the populations

of non-parasitic nematodes in the root.

Figures 7: Number of non-parasitic nematodes in 5g of root of olive trees treated with two


Alamein-Pisco-Ica)

0 .0 0

1 0 0 .0 0

2 0 0 .0 0

3 0 0 .0 0

4 0 0 .0 0

5 0 0 .0 0

6 0 0 .0 0

41

In the third sampling the Control treatment, alone, had an increase in population with 242

nematodes in 5g of root. The other treatments maintain populations similar to prior populations,

affirming that there was a nematicidal effect that avoided a nematode increase in the root.

In the fourth sampling in spite of being statistically similar all the treatments had a

significant increase in populations due to higher temperatures, the highest proportional increase

was in the HUNTER treatment. The Control treatment was the only one to have lower

populations due to some external factor.

In the fifth sampling all the treatments are statistically similar, however, the treatment

with BIO-BAC had a much smaller population that the other treatments, being similar to the

initial population. Also having a lower population of non-parasitic nematodes in the root was the

treatment with HUNTER showing there was a control on the part of the nematicides. The

treatments with HUNTER+BIO-BAC, ALDICARB and CONTROL had population increases

with the Aldicarb treatment having the highest populations (525 in 5g of soil).

4.3 POPULATION OF OTHER NEMATODES FOUND IN THE SOIL

4.3.1 Population Fluctuations of Aphelenchoides in 100 cc of soil in olive tree

Table 13 and Figure 8 show, in general, that the population of Aphelenchoides is nil in all

the treatments throughout the whole experiment.

This could indicate that this nematode is in very low populations in the soil and would

not be causing damage to the roots or foliage and that the action of the nematicides is efficient in

the control, maintaining the low populations of this nematode and avoiding an increase.

Table 13: Number of Aphelenchoides in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

Treatment 1st Sample 2nd Sample 3rd Sample 4th Sample 5th SampleHUNTER 0 0 0 0 0BIO-BAC 0 1 0 0 0HUNTER+BIO-BAC 0 0 0 0 0ALDICARB 0 0 0 0 0

42

CONTROL 0 1 0 0 0

Figure 8: Number of Aphelenchoides in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

0

0 . 2

0 . 4

0 . 6

0 . 8

1

1 . 2

1 2 3 4 5

T i m e

Num

ber o

f Ind

ivid

uals

H u n t e r B i o - B a c H u n t e r - B i o - B a c A l d i c a r b C o n t r o l

4.3.2 Population fluctuations of Aphelenchus in 100 cc of soil of olive tree cultivation

Table 18 and Figure 9 shows a defined trend in the behavior of the population of

Aphelenchus.

In general, the population in the treatments with HUNTER, HUNTER+BIO-BAC and

ALDICARB were lower until the third sampling. That was not observed in the treatments with

BIO-BAC and CONTROL, where the population increased through the second sampling then

diminished in the third sampling finally having populations similar to the initial population. In

the fourth sampling, while the population in the treatment with HUNTER continued to decline,

the other treatments had an increased population. In the fifth sampling only the treatment with

BIO-BAC reduced the population to zero. In the other treatments the populations increased or

stayed similar to the previous sampling.

Picture 14: Number of Aphelenchus in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

Treatment 1st Sample 2nd Sample 3rd Sample 4th Sample 5th Sample

43

HUNTER 9 6 5 3 13BIO-BAC 8 15 8 18 0HUNTER+BIO-BAC 9 3 0 4 5ALDICARB 5 5 3 8 11COTROL 1 10 3 8 8

In general, the best treatment in the control of Aphelenchus was HUNTER+BIO-BAC,

followed by the treatment with HUNTER, ALDICARB, CONTROL and lastly the treatment

with BIO-BAC that in spite of having a population of zero in the previous samplings had

populations higher than the other treatments.

44

Figure 9: Number of Aphelenchus in 100 cc of soil in olive trees treated with two biological

nematicides, Aldicarb and untreated in fields infested nematodes (Orchard Alamein-Pisco-

Ica)

0

2

4

6

8

1 0

1 2

1 4

1 6

1 8

2 0

1 2 3 4 5

T i m e

Num

ber o

f Ind

ivid

uals

H u n t e r B i o - B a c H u n t e r + B i o - B a c A l d i c a r b C o n t r o l

4.3.3 Population Fluctuations in Ditylenchus nematodes in 100 cc of soil of olive trees

Table 15 and Figure 10 indicate that, in general, populations of Ditylenchus are nil in all

the treatments throughout the whole experiment.

45

Table 15: Number of Ditylenchus in 100 cc of soil in olive trees treated with two biological

nematicides, Aldicarb and untreated in fields infested with nematodes (Orchard Alamein-

Pisco-Ica)

Treatment 1st Sample 2nd Sample 3rd Sample 4th Sample 5th SampleHUNTER 0 0 0 0 0BIO-BAC 0 0 0 0 0HUNTER+BIO-BAC 0 0 0 1 0ALDICARB 0 0 0 0 0CONTROL 0 0 0 0 0

46

Figure 10: Number of Ditylenchus in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)

0

0 . 2

0 . 4

0 . 6

0 . 8

1

1 . 2

1 2 3 4 5T i m e

Num

ber o

f Ind

ivid

uals


The above is an indication that this nematode is in very low numbers in the soil and

would not be causing damage to the roots. Also, the action of the nematicides is efficient in the

control of this nematode, maintaining the low populations and avoiding an increase..

4.3.4 Fluctuations in population of Helicotylenchus in 100 cc of soil in olive trees

Table 16 and Figure 11 indicate that the populations of Helicotylenchus are almost nil in

all the treatments throughout the experiment. This could be an indication that the nematode is in

very low populations in the soil and would not be causing damages to the roots. Also, the action

of the nematicides is efficient in the control of this nematode, maintaining low populations and

avoiding an increase.

Table 16: Number of Helicotylenchus in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)


47

Figure 11: Number of Helicotylenchus in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

0

1

2

3

4

5

6

1 2 3 4 5

T i m e

Num

ber o

f ind

ivid

uals


In spite of the population drop, the best treatment in the control of this nematode is

ALDICARB, which maintained a zero population throughout the experiment, followed by the

treatments with HUNTER+BIO-BAC and CONTROL which were similar, and then the BIO-

BAC and HUNTER.

48

4.3.5 Fluctuations in the population of Hemicycliophora in 100 cc of soil in the

cultivation of olive trees

Table 17 and Figure 12 show a similar behavior in all the treatments except in the

ALDICARB where the population stayed at zero throughout the experiment.

In the treatments with HUNTER, BIO-BAC and CONTROL populations increase in the

second sampling, whereas, in the treatment with HUNTER+BIO-BAC the population decreases.

In the third sampling HUNTER, BIO-BAC decreased populations and CONTROL decreased the

populations to zero then maintained that level throughout the rest of the experiment. The

treatment with HUNTER+BIO-BAC had an increase in population. In the fourth sampling the

treatments with HUNTER and HUNTER+BIO-BAC decreased populations, in HUNTER

populations stayed at 3 in 100 cc of soil until the end of the experiment. HUNTER+BIO-BAC

increased populations to the point of equaling the initial population. In the case of the BIO-BAC

the final population fell to zero.

The best treatment in the control of this nematode was ALDICARB in that it didn't allow

an increase in the populations throughout the experiment. The next best treatments, in order,

were HUNTER+BIO-BAC, BIO-BAC, CONTROL and lastly HUNTER.

Table 17: Number of Hemicycliophora in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)


Figure 12: Number of Hemicycliophora in 100 cc of soil in olive trees treated with two


Alameitn-Pisco-Ica)

49

0

5

1 0

1 5

2 0

2 5

3 0

1 2 3 4 5

T i m e

Num

ber o

f Ind

ivid

uals


4.3.6 Population fluctuations of Pratylenchus in 100 cc of soil in the cultivation of olive

tree

Table 18 and Figure 13 show a reduction in the populations, due to the effect of the

nematicides.

Table 18: Number of Pratylenchus in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)


Similar values occurred in all the treatments in the initial population. This indicated that

the nematode is present and and dispersed evenly throughout the whole field. In the second

sampling there is a decrease in the populations in all the treatments due to the effect of the

nematicides, with values at zero in HUNTER, BIO-BAC and ALDICARB treatments. In the

treatment with BIO-BAC the populations stay at zero until the end of the experiment. In the

other treatments, with the exception of HUNTER+BIO-BAC, populations stay at zero or at a

50

value very near zero throughout the experiment. In the HUNTER+BIO-BAC treatment, the final

population increased to 34 in 100 cc of soil.

The best treatment in the control of Pratylenchus was BIO-BAC, followed by

ALDICARB, very close was HUNTER, then the CONTROL and lastly the treatment with

HUNTER+BIO-BAC. The mixture of biological nematicides did not show a synergistic effect,

possibly because the active ingredients of the products are incompatible and alone each of them

would be acting on the nematodes. It is noted that a lesser effect occurred than that of the

products applied separately.

51

Figure 13: Number of Pratylenchus in 100 cc of soil in olive trees treated with two biological

nematicides, Aldicarb and untreated, in fields infested with nematodes (Orchard Alamein-

Pisco-Ica)

0

5

1 0

1 5

2 0

2 5

3 0

3 5

4 0

1 2 3 4 5T i m e

Num

ber o

f Ind

ivid

uals

H u n t e r B io - B a c H u n t e r + B io - B a c A ld i c a r b C o n t r o l

4.3.7 Population fluctuations of Rotylenchus in 100 cc of soil in the cultivation of olive

Table 19 and Figure 14 show the populations of Rotylenchus are almost nonexistent in all

the treatments throughout the experiment, except for the control that has a count of 88 in 100cc

of soil in the fourth sampling.

Table 19: Number of Rotylenchus in 100 cc of soil in olive trees treated with two biological

nematicides, Aldicarb and untreated, in fields infested with nematodes (Orchard Alamein-

Pisco-Ica)


As indicated, this nematode is in very low numbers in the soil and would not be causing

damage to the roots. The action of the nematicides is efficient in controlling this nematode,

maintaining a low population and avoiding an increase.

52

Figure 14: Number of Rotylenchus in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)

0

1 0

2 0

3 0

4 0

5 0

6 0

7 0

8 0

9 0

1 0 0

1 2 3 4 5T i m e

Num

ber o

f Ind

ivid

uals


Comparing treatments, the smallest population of nematodes is HUNTER and BIO-BAC,

followed by HUNTER+BIO-BAC, ALDICARB and lastly CONTROL.

4.3.8 Fluctuations in the population of Tylenchorhynchus in 100 cc of soil in the

cultivation of olive tree

Table 20 and Figure 15 show that the populations stay very similar throughout the

experiment.

53

Table 20: Number of Tylenchorhynchus in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)


In the first sampling all the populations are similar (an average of 14 per 100 cc of soil)

with the exception of the CONTROL that had a population of 3 per 100 cc of soil. In the second

sampling all the treatments had a slight increase in population, except for the HUNTER

treatment that maintained a population similar to that of the initial population. In the third

sampling the populations of the treatments with BIO-BAC, ALDICARB, and HUNTER+BIO-

BAC decreased, while the HUNTER and CONTROL treatments increased. In the fourth

sampling all the treatments had population decreases, in the fifth sampling populations increased

in all the treatments to levels higher than the initial population.

Figure 15: Number of Tylenchorhynchus in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

0

1 0

2 0

3 0

4 0

5 0

6 0

1 2 3 4 5T i m e

Num

ber o

f Ind

ivid

uals


54

The above shows that the best treatment for the controlled of this nematode was BIO-

BAC, followed by ALDICARB, CONTROL, HUNTER and lastly HUNTER+BIO-BAC.

4.3.9 Population fluctuations of Tylenchus in 100 cc of soil in olive trees

Table 21 and Figure 16 shows the populations remain low and without uniform behavior

among the treatments throughout the experiment.

In the first sampling the populations are not uniform among the treatments, an advantage

is observed in HUNTER and CONTROL treatments that have zero populations. In the second

sampling the HUNTER and CONTROL have population increases, with HUNTER having a

smaller increase than CONTROL. In the other treatments the populations decrease in some cases

to zero such as in the treatments with HUNTER+BIO-BAC and ALDICARB.

Table 21: Number of Tylenchus in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)


In the third sampling populations in the treatments with HUNTER and BIO-BAC

continued to decrease until they reach zero, the CONTROL treatment also attained a zero

population that it maintained throughout the remainder of the experiment. In the fourth sampling

the populations in the HUNTER and BIO-BAC treatments again increased, the treatments with

HUNTER+BIO-BAC and ALDICARB had lower populations. In the fifth sampling HUNTER

and BIO-BAC treatments had lowered populations, whereas, HUNTER+BIO-BAC and

ALDICARB had increases.

Figure 16: Number of Tylenchus in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)

55

0

2

4

6

8

1 0

1 2

1 2 3 4 5T im e

H u n te r B io - B a c H u n te r + B io - B a c A ld ic a r b C o n t r o l

The treatment that controlled this nematode best was BIO-BAC, followed by HUNTER,

CONTROL, ALDICARB and lastly HUNTER+BIO-BAC.

4.3.10 Population fluctuations of Xiphinema in 100 cc of soil in the cultivation of olive

tree

Table 22 and in Figure 17 show that the populations of Xiphinema are almost nil in all the

treatments throughout the experiment.

Table 22: Number of Xiphinema in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)


Figure 17: Number of Xiphinema in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)

56

0

0 . 2

0 . 4

0 . 6

0 . 8

1

1 . 2

1 2 3 4 5T i m e

Num

ber o

f Ind

ivid

uals


The above indicates that this nematode is in very low populations in the soil and was not

causing damage to the roots. The action of the nematicides is efficient in the control of this

nematode maintaining the low populations.

4.3.11 Population fluctuations of Criconematidos in 100 cc of soil in the cultivation of

olive trees

Table 23 and Figure 18 shows the ALDICARB and CONTROL treatments had declines

while the other treatments had no defined tendencies.

ALDICARB and CONTROL treatments had a population increase in the second

sampling, which stopped, then diminished progressively until the final population, ending up at

zero in both cases.

In the treatment with BIO-BAC a decrease is observed from the beginning of the

experiment until the third sampling, it then increases through the fourth sampling and diminishes

again in the final population, having a lower value than the initial population.

The HUNTER and HUNTER+BIO-BAC treatments had populations increase in the

second sampling that then declined in the third sampling, then leveling off. In the fourth

sampling an increase in population occurred in the HUNTER treatment whereas a decline

continued in the HUNTER+BIO-BAC. In the fifth sampling the treatment with HUNTER, again,

had a decline in populations to 5 per 100 cc of soil, which was smaller than the initial population.

The HUNTER+BIO-BAC had a population increase up to 25 per 100 cc of soil, larger than the

initial population.

57

Table 23: Number of Criconematidos in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)


58

Figure 18: Number of Criconematidos in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

0

5

1 0

1 5

2 0

2 5

3 0

3 5

4 0

1 2 3 4 5

T i m e

Num

ber o

f Ind

ivid

uals

H u n t e r B io - B a c H u n t e r + B io - B a c A ld ic a r b C o n t r o l

The treatment with the smallest population of Criconematidos is the CONTROL,

possibly due to a deterioration of the quality and quantity of the roots. The next best treatment is

ALDICARB, followed by the treatments with BIO-BAC, HUNTER and HUNTER+BIO-BAC.

4.3.12 Population fluctuations of Dorylaimidos in 100 cc of soil in the cultivation of olive

In table 24 and Figure 19 an upward trend, similar in all the treatments is observed.

Table 24: Number of Dorylaimidos in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)


59

Figure 19: Number of Dorylaimidos in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

01 02 03 04 05 06 07 08 09 0

1 0 0

1 2 3 4 5

T i m e

Num

ber o

f Ind

ivid

uals


Except for the ALDICARB, whose population started to decline in the third sampling and

ended with a final population of 38in 100 cc of soil, populations increased until the fourth

sampling. In the fifth sampling the populations declined, but ended up higher than the initial

population.

The treatment with the smallest population of Dorylaimidos was the CONTROL,

followed, in order, by the treatments with HUNTER+BIO-BAC, ALDICARB, HUNTER and

lastly the BIO-BAC. The populations of these nematodes are not very important because they

don't attack the roots of the olive tree.

4.3.13 Population fluctuations of Mononchidos in 100 cc of soil in the cultivation of

olive tree

Table 25 and Figure 20 show, in general, that the populations of Mononchidos are almost

nil in all the treatments throughout the experiment.

60

Table 25: Number of Mononchidos in 100 cc of soil in olive trees treated with two biological

nematicides, Aldicarb and untreated in fields infested of nematodes (Orchard Alamein-

Pisco-Ica)


The Table indicates that this nematode is in very low populations in the soil and would

not be exercising a natural control on the phytoparasitic nematode.

4.3.14 Population fluctuations of Rhabditidos in 100 cc of soil in olive tree cultivation

Table 26 and Figure 21 show that the treatments with BIO-BAC and CONTROL are

similar and different from the other treatments that has similar results among them.

CONTROL first had a population increase then slight increases are observed followed by

a low population, however, the populations stayed almost the same until the end of the

experiment. With BIO-BAC a decrease in populations occurs until the third sampling where it

levels off then stays similar until the end of the experiment, similar to the populations of

CONTROL.

Figure 20: Number of Mononchidos in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

0

1

2

3

4

5

6

1 2 3 4 5

T i m e

Num

ber o

f Ind

ivid

uals


61

Table 26: Number of Rhabditidos in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)


In the treatment with HUNTER and ALDICARB the populations first increase and then

diminish. Treatments with HUNTER+BIO-BAC first had a decline then an increase, then leveled

off. In the third sampling the three treatments have similar populations that remain this way until

the end of the experiment. At the end of the experiment, comparing different behaviors, the

populations of the five treatments are similar to each other and similar to the first sampling.

In general, the best treatment in the control of Rhabditidos, taking into consideration that

these nematodes are not phytoparasites, is the CONTROL, followed by BIO-BAC,

HUNTER+BIO-BAC, HUNTER and lastly the ALDICARB. A high number of Rhabditidos in

the soil indicates a lesser effect for the products on the beneficial nemato-fauna with more

biodiversity and biological activity that has a negative effect on phytoparasitic nematodes.

Figure 21: Number of Rhabditidos in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

1 2 3 4 5T i m e

Núm

ero

de in

divi

duos


62

4.3.15 Population fluctuations of Trichodoridos in 100 cc of soil in olive tree cultivation

Table 27 and Figure 22 show that the populations of Trichodoridos are almost nil in all

the treatments throughout the experiment, except for the second sampling where a spike is

observed in the populations of all the treatments except for the HUNTER.

Table 27: Number of Trichodoridos in 100 cc of soil in olive trees treated with two biological


Pisco-Ica)


The above table indicates that this nematode is in very low populations in the soil and it

would not be causing damages to the roots, and that the action of the nematicides is efficient in

the control of this nematode maintaining low populations and avoiding increases.

The treatment with the lowest number of this nematode is HUNTER, followed by

CONTROL, BIO-BAC, ALDICARB and lastly HUNTER+BIO-BAC.

Figure 22: Number of Trichodoridos in 100 cc of soil in olive trees treated with two


Alamein-Pisco-Ica)

0

2

4

6

8

1 0

1 2

1 4

1 2 3 4 5

T i e m p o

Num

ber o

f Ind

ivid

uals


63

4.4 YIELD

Due to the abnormal climatic conditions caused by El Nino, in comparison to normal

years, the yield that was very low.

Table 28: Table of yields of olive trees treated with two biological nematicides, Aldicarb

and untreated in fields infested with root knot nematode (Orchard Alamein-Pisco-Ica)

Treatment Fruit Weigh(g)

Fruits/Tree(n°)

Yield/Tree(g)

HUNTER 9.1 ab 22 b 204 bBIO-BAC 9.2 ab 38 a 347 aHUNTER+BIO-BAC 9.0 b 20 c 180 cALDICARB 9.2 a 18 d 161 dCONTROL 6.0 c 17 d 102 e

Attachment 5 contains the various characteristic data of yield that were used in the

analysis of variance the results of which are given in Table 28.

4.4.1 YIELD PER TREE

Table 29 and Figure 23 shows that the CONTROL is inferior to all the other treatments.

The increase in the yield is more than 57% in all the cases. The plants treated with BIO-BAC had

a yield increase of 239% compared to the control that had a yield of 102g per tree.

Table 29: Yield average, in grams, for tree of olive tree treated with two biological

nematicides, Aldicarb and untreated in fields infested with root knot nematode (Orchard

Alamein-Pisco-Ica)

Treatment Rep. 1 Rep. 2 Rep. 3 Rep. 4 Aver. Incr.CONTROL 103 103 100 104 102 -HUNTER 198 212 194 214 204 99%BIO-BAC 355 338 353 340 347 239%HUNTER+BIO-BAC 171 180 198 173 180 76%ALDICARB 160 158 166 159 161 57%

The basis of this data was subjected to analysis of variation, in which statistical differences

were detected at a level of probability of 0.05. This is shown in Attachment 13, where it is

64

observed that all the treatments are significantly different and larger than the CONTROL.

Having a highest value were the plants treated with BIO-BAC with 347g, followed by HUNTER,

the mixture of HUNTER+BIO-BAC and ALDICARB, with the CONTROL having 102g.

The plants treated with HUNTER didn't show the best results in population fluctuations

of nematodes in soil and roots. Their yields, keeping in mind that they were not normal, were

among the highest due to the biostimulant effect of this product. It causes a more new root

formation that won't be attacked by the nematodes and that will allow an increased absorption of

nutrients that translated in a higher yield. The treatment with BIO-BAC had a decreased

population in the root allowing a good absorption of nutritious and a higher yield (the highest).

As for the treatments with ALDICARB and HUNTER+BIO-BAC, the progressive increase of

nematodes in the roots and in the soil led to deterioration of root systems. This led to a low

absorption of nutritious and a low yield, but more than the CONTROL. This is because these

products protected the roots for a period of time allowing a good absorption of nutritious during

this interval, obtaining a better yield than the CONTROL.

Figure 23: Yield average per tree of olive tree treated with two biological nematicides,

Aldicarb and untreated in fields infested with root knot nematode (Orchard Alamein-

Pisco-Ica) in grams

0.00

50.00

100.00

150.00

200.00

250.00

300.00

350.00

400.00

Treatm ent

Yiel

d (g

r)

H unter B io-B ac H unter+B io-B ac A ld icarb C ontro l

According to ANPEAP (3), the normal average yearly yield per olive tree is 42 kg when

it has chilling temperatures (approximately a 10-day period of cold during June and July of less

than 12°C). These temperatures act as a "trigger" that make the floral buds develop, they fill and

open up. This didn't happen during the development of the present work, as is shown in the Table

65

3. This caused very low flowering, fruit parthenocarpy formation that doesn’t develop and low

fruiting, which leads to low yields.

4.4.2 WEIGH OF FRUIT

Table 30 and Figures 24 shows that an increase in the weight of fruit occurred due to the

application of nematicides. All the treated plants have a 40% higher fruit weight compared to the

control and the plants treated with BIO-BAC increased by 52% compared to CONTROL that had

an average fruit weight of 6.0g.

Table 30: Average weigh of fruit of olive tree treated with two biological nematicides,


Pisco-Ica) in grams

Treatment Rep. 1 Rep. 2 Rep. 3 Rep. 4 Aver. Incr.CONTROL 6.0 6.1 5.9 6.1 6.0 -HUNTER 9.0 9.2 9.0 9.1 9.1 51%BIO-BAC 9.3 9.0 9.3 9.1 9.2 52%HUNTER+BIO-BAC 9.0 9.0 9.0 9.1 9.0 50%ALDICARB 9.2 9.3 9.2 9.1 9.2 53%

The data it was subjected to the analysis of variation in which showed statistical

differences at a level of probability of 0.05.

Figure 24: Average weigh of fruit of olive tree treated with two biological nematicides,


Pisco-Ica) in grams

0.001.002.003.004.005.006.007.008.009.00

10.00

Treatm ent

Wei

ght (

gr)

H un te r B io -B ac H un te r+B io -B ac A ld ica rb Testigo

66

According to Attachment 14 the products used have a positive effect since the fruit

weights are significantly different from CONTROL. Plants treated with ALDICARB, BIO-BAC

and HUNTER have the best fruit weight, being 9.2, 9.2 and 9.1 grams respectively. Plants with

the HUNTER+BIO-BAC treatment at 9.0 g are in the second group along with the treatments

BIO-BAC and HUNTER. Finally the CONTROL is statistically smaller than all the others.

4.3.3 NUMBER OF FRUIT

Table 31 and Figure 25 show that CONTROL is lowest of all the treatments. Increases in

the number of fruit occurred in all cases, with the biggest increase (122%), compared to

CONTROL (that produced 17 fruits per tree) were those trees treated with BIO-BAC.

This data was subjected to the analysis of variance in which statistical differences were

detected at a level of probability of 0.05.

Table 31: Average Number of fruit per olive tree treated with two biological nematicides,


Pisco-Ica)

Treatment Rep. 1 Rep. 2 Rep. 3 Rep. 4 Aver. Incr.CONTROL 17 17 17 17 17 -HUNTER 22 23 22 24 22 33%BIO-BAC 38 38 38 37 38 122%HUNTER+BIO-BAC 19 20 22 19 20 18%ALDICARB 17 17 18 18 18 3%

Figure 25: Average Number of fruit per olive tree treated with two biological nematicides,


Pisco-Ica)

0 .0 0

5 .0 0

1 0 .0 0

1 5 .0 0

2 0 .0 0

2 5 .0 0

3 0 .0 0

3 5 .0 0

4 0 .0 0

T re a tm e n t

Num

ber o

f fru

it

H u n te r B io -B a c H u n te r+ B io -B a c A ld ic a rb C o n tro l

67

Attachment N° 15 shows that there are significant differences with a coefficient of

variation of 3.47%, therefore the Duncan test was performed at the 0.05 level of probability. This

showed that the biologic products BIO-BAC, HUNTER and the mixture of HUNTER and BIO-

BAC had a positive effect on the number of fruit and they are statistically different from the

CONTROL. The trees treated with ALDICARB are statistically similar to the control.

V. CONCLUSIONS

1. The best treatment in nematode population reduction and yield was BIO-BAC. Since, out of

ten analyzed characteristics (number of juvenile of M. incognito in 100 cc soil and in 5g of

root, index of nodulation caused by M. incognito, number of parasitic nematode in 100 cc

soil, in 5g of root, number of non-parasitic nematodes in 100 cc soil, and in 5g of root,

weight of fruit in grams, number of fruits per tree and yield per tree in grams) it ranked first

out of seven. Because the action of the fungi and bacteria worked to control this nematode

and the presence of micronutrients, plants developed better. The treatment with

HUNTER+BIO-BAC ranked first in the nodulation index, the CONTROL ranked first in the

number of non-parasitic nematodes in the soil and the ALDICARB treatment ranked first in

fruit weight (in grams).

2. The biological products controlled nematodes and reduced their root damaging effects, as

evidenced in the soil and nodulation index comparison with the other treatments.

3. The ALDICARB treatment reduced the population of nematodes but after losing its residual

effect the population increased to above initial levels. This resulted in an increase in the

index of nodulation that diminished the vigor and normal development of the roots and

therefore reduced production, but not as low as CONTROL.

4. Although HUNTER had high populations in the soil, the damage caused by M. incognito in

the root system of the plants was light. This was because of the mode of action of the product

is to cause a nematistatic effect, causing neuromuscular alterations in activities like

movement, feeding and other sensorial aspects. Also, this product causes metabolic changes

in the plants and provides micronutrients allowing it to grow strong prior to nematode attack

and favoring a considerable increase of new roots.

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5. There were no synergistic effects observed in the HUNTER+BIO-BAC treatment. It was the

best treatment in only one of the characteristics (nodulation index); in no other cases was it

better than the BIO-BAC alone, but it was, in some cases, better than HUNTER (number of

juvenile M. incognito in 100 cc soil and in 5g of root, nodulation index caused by M.

incognito, number of parasitic nematode in 100 cc soil, in 5g of root, number of non-parasitic

nematodes in 100 cc soil, but in none the yield parameters), possibly because some of the

active ingredient of the products competed with each other and did not work efficiently.

6. Although the population of nematodes in the soil in the CONTROL treatment was not very

high, the nodulation index was, causing a decrease of plant vigor and normal development,

which resulted in a drastic reduction in production.

7. All the biological treatments were better than the chemical treatment and CONTROL in

yield, both in the number of fruit per tree and in weight per fruit. The best was BIO-BAC that

had increases in weight per fruit (52%), number of fruit (122%) and yield (239%), compared

to CONTROL.

8. High temperatures during the experiment and the radical decrease in temperature at the end,

along with external factors such as watering favored the unstable behavior of nematodes and

therefore favored the migration of nematodes causing highly varied populations in the soil.

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VI. RECOMMENDATIONS

1. To repeat this experiment at least two years more, to corroborate the results of the nematode

populations obtained in this work, and to better observe the effect of the nematicide products

on yield, since the El Nino Phenomenon had an abnormal effect on the data.

2. Using the results of this work as a basis, carry out tests to optimize rates and dates of

application.

3. To carry out this same study in other areas to compare the results with different

environmental factors, types of soil, and cultivation management.

4. Because HUNTER is a slow acting product, to apply it in combination with organic matter or

with some other biological control.

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SUMMARY

An experiment was carried out to study the effect of two biological nematicides and

Aldicarb on the populations of root knot nematode (M. incognito) and on the yield of olive trees.

The trial was carried out in field 8 of the Alamein Orchard, located in the county of Pisco in the

Ica area, in 40-year-old olive groves of the variety Sevillana, during the months of September

through June of 1999.

A statistical randomized block design was used throughout, with 4 repetitions and the

following treatments: HUNTER (15 cc/plant), BIO-BAC (20 cc/plant), HUNTER+BIO-BAC (15

cc + 20 cc/plant), Aldicarb (200 g/plant) and a CONTROL without application of any product. 3

applications of the biologic products (HUNTER, BIO-BAC, and HUNTER+BIO-BAC) and 1 of

Aldicarb were made. There were 5 samplings, 1 before the application of the products, and the

remaining samplings, 30 to 45 days after each application.

Evaluations made were as follows: number juvenile in 100 cc of soil; number juveniles

and eggs in 5g of root; the index of root nodulation in tomatoes (bioassay); number of parasitic

and non-parasitic nematodes in the soil and root; yield per tree; number of fruit per tree and

weight of each fruit.

The results obtained demonstrated that BIO-BAC was better in controlling nematodes in

the soil and in the root and had the best yield, which was 347g per tree. This nematode control

maintained an almost constant nematode population level, avoiding, in many cases, having to

overcome the levels of initial population. Of the ten evaluated characteristics (number of

juveniles of M. incognito in 100 cc of soil and in 5g of root, index of nodulation caused by M.

incognito, number of parasitic nematodes in 100 cc of soil and in 5g of root, number of non-

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parasitic nematode in 100 cc of soil and in 5g of root, weight in grams of fruit, number of fruit

per tree and yield, in grams, per tree) it rated best in seven.

A lesser effect on nematode control was obtained with HUNTER+BIO-BAC, HUNTER,

ALDICARB and lastly the CONTROL, which showed no effect on the nematode populations. A

lesser effect was also obtained in yield with the treatments where HUNTER at 204g per tree,

HUNTER+BIO-BAC at 180g per tree and ALDICARB at 161g per tree; whereas, the

CONTROL had a yield of 102g per tree.

effect of two biological nematicides and aldicarb … · national agrarian university of molina...

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