plant-based pesticides for the control and...
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PLANT-BASED PESTICIDES FOR THE CONTROL AND MANAGEMENT OF
SELECTED PEST FOR ORGANIC VEGETABLE PRODUCTION IN ILOCOS
Leticia A. Lutap, Lucricia Conchita G. Cocson, Rodalyn G. Quijano,
Agustin A. Quidilla, Alecsis G. Villarin and Aida D. Solsoloy
Abstract
With the increasing concern for environmental safety and human health,
development of alternative control methods for crop production such as the use of
biopesticides against major pests of vegetable crops is a necessity. Such crop insect
pests were tomato fruitworm, Helicoverpa armigera Hubn.,thrips (Thrips tabaci),
mites (Aceria tulipae) and Epilachna beetle (Epilachna vigintioctpucntata) and
aphids ( A.cracivora) while on diseases were Alternaria solani causing early blight
on tomato, Alternaria porri, causing purple blotch and Cercospora duddiae causing
cercospora leaf spot on garlic, respectively. Plants with known pesticidal properties
were collected and re-evaluated as biopesticides.
Pesticidal effect on target pests was noted from plants such as Cleome
viscosa, Argemone mexicana, Euphorbia hirta, Tabernaemontana pandacaqui.
Cucurma longa, Origanum vulgare. Piper betle, Lantana camara, Allium sativum,
Aloe barbadensis Azadirachta indica leaves and garlic waste Insect growth
inhibitory effect was observed such as reduced number of larval and pupal days, as
well as premature mortality of treated larvae. Using the formulated products, the
effectiveness was comparable with chemical pesticides under field conditions; lower
disease intensity and higher marketable yield were noted compared to farmers
practice. Cost and return analysis also showed that the different products is
comparable with chemical pesticides. Microbial antagonists isolated from goat
manure tea and bat dung were identified and proved effective in vitro against A.
solani in tomato and A. porri and C. duddiae in garlic. Shelf life of the products
showed potency after six months of storage. Results mentioned proved that the
formulated biopesticides were very essential for organic vegetable production in the
Ilocos.
_________________________________________________________________
Keywords: Plant extracts, pesticidal property, toxicity, product formulation,
insectidal effect, insect pests, disease–causing organism, antifungal
activities, disease severity
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B. TECHNICAL REPORT
DEVELOPMENT OF PEST MANAGEMENT PRODUCTS AND SYSTEMS FOR
ORGANIC VEGETABLE PRODUCTION IN ILOCOS REGION
Leticia A. Lutap, Lucricia Conchita G. Cocson, Rodalyn G.Quijano,
Agustin A. Quidilla, Alecsis G. Villarin and Aida D. Solsoloy
INTRODUCTION
With the rising concern for environmental safety and human health, regaining interest in
the development of alternative control methods should be given proper priority and attention,
among which is the use of bio-pesticide. The bio-pesticide science is still evolving; hence in
depth research is needed in many areas such as production formulation, delivery and
commercialization of these products.
Plants are composed of chemical substances of which some are not directly beneficial to
the growth and development of the organism. These secondary compounds have been regarded
as part of the plant’s defense mechanism against plant feeding insects and other herbivores
(Rosenthal and Janzen, 1979). These compounds have different properties such as attractants,
ovicides, insecticides, and anti-feedants (Muruganm et al., 1978). Therefore searching for new
alternative control methods using plants with pesticidal properties can be exploited either as
powder or as crude extract in water or other organic solvents. The challenge is to develop a
formulation and application method that can be implemented on a commercial scale that is
effective, reliable, consistent and economically feasible.
In the Ilocos region, tomato, garlic, eggplant, sweet and finger pepper are vegetables
usually planted after rice. In addition to being considered as cash crops, they are also rich sources
of vitamins and minerals. However, like any other crop, they are being attacked by insect pests
and diseases, a major constraint in production. For example, tomato is attacked by the tomato
fruitworm Helicoverpa armigera. Although foliage feeding of the newly hatched larvae in
tomato may not cause any significant damage, boring may result in yield loss up to 70%. Thrips
and mites, which are known vectors of viruses likewise cause damage to sweet and finger
peppers. High population of these pests produces leaf curl, distortion of plant growth lower
deformation, and wilting. Epilachna beetle, Epilachna vigintioctpucntata F), is now becoming a
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major pest in eggplant and the most prevalent pest especially toward the onset of the rainy
seasons. Both the larva and the adult damage the plant by eating the leaf tissues between the
veins and skeletonized leaves dry out. Also tangle top infestation due to mites Aceria tulipae can
reduce production by as much as 40-50% (Ahmed 1981) and thrips by as much as 50% (Ligsay
et al. 1984).
On the other hand, the industry of the Ilocos white garlic, a much preferred one over the
imported Taiwan variety due to its pungency and aroma, is slowly collapsing due to the low
yield. This low yield is accounted to disease attack particularly purple blotch caused by
Alternaria porri (Ellis) Cif and cercospora leaf spot caused by Cercospora duddiae Welles.
While tomato production is being beseeched by the early blight caused by Alternaria
solani (Ellis and Martin) Jones and Grout. This disease is favored by hot and humid condition
and if not controlled can cause 90-100% yield loss.
Generally, farmers use synthetic chemical pesticides to abate pest incidence in the
absence of alternatives. Its use has been an important part of pest management for many years;
however with known disadvantages and risk; some synthetic pesticides leave unwanted residues
in food, water and the environment. Some are suspected carcinogens and low doses of many
insecticides and fungicides are toxic to mammals. This predicament results in the search for less
hazardous alternatives to conventional synthetic insecticides. Among the recent efforts is the
exploitation of natural products from plants that contain toxic metabolites or the use of microbial
antagonists to control diseases. Botanicals degrade more rapidly than most chemicals pesticides
and are therefore considered relatively environmental friendly and less likely to kill beneficial
pests than synthetic pesticides with longer environmental retention. It is therefore an utmost
urgency to identify alternative to chemical pesticide in plant protection. Biopesticides which are
target- specific, eco-friendly and biodegradable can be a potential alternative. Hence, different
plants using extracts at different formulation were investigated.
OBJECTIVES
General:
To screen different plant materials with potential pesticidal activity specifically
insecticidal and fungicidal action on pests of selected crops such as tomato, garlic, finger and
sweet pepper, eggplant.
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Specific:
1. To formulate the most effective plant materials and biocontrol agents as bio-pesticide
products against pests of priority vegetable crops;
2. To determine the effectiveness of the formulated bio-pesticide products against test insect
pests and diseases under laboratory, greenhouse and field conditions; and
3. To generate data for possible registration to the Fertilizer and Pesticide Authority
4. To develop a package of technology with the products as main component particularly in
an organic vegetable production system.
REVIEW OF LITERATURE
A global Natural Product Database, Napralert, is continuously produced by College of
Pharmacy, University of Illinois, Michigan, United States and is said to cover all published
material on plants with pesticidal properties. Linking the information obtained from the database
to the knowledge of the indigenous flora could draw rings on the most promising plants which
deserve further studies. Important aspects to consider are not only pesticidal properties of the
plants but also their distribution, abundance and easiness to be propagated. (Rees et al. 1993).
The mechanisms behind an extract’s pesticidal properties are of course not always
understood. Nor are they easy to observe in the field. Bioasay with crude or purified extracts
could contribute information about insect behavior and development and thus facilitate an
interpretation of the results. Anti-feedant activity assay with leaf disks in a choice situation has
been used at ICIPE and is described by Kubo (1993).
The possibility for small-scale farmers to buy agricultural inputs is limited. This together
with the problems of health risks and environmental pollution, owing to misuse of chemicals,
provide good arguments for carrying out studies on natural pesticides. Support to these activities
is therefore strongly recommended with particular reference to pest problems in vegetables,
stored products and tree plantations. The aim should be develop reliable pest control methods
which are attractive and safe for farmers to use.
The biochemical pesticides include plant extracts, pheromones, plant hormones, natural
plant-derived regulators, clay, potassium bicarbonate and enzymes as the active ingredients.
Biopesticides are used primarily as preventive measures, so they may not perform as quickly as
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some synthetic chemical pesticides. However, biopesticides are generally less toxic to the user
and non-target organisms, making them desirable and sustainable tools for disease management.
Due to indiscriminate use of chemical pesticides, H. armigera has developed resistance to
some insecticides at several locations in the country. Therefore, there is a need to find out
suitable plant-based pesticides. In the present study, extracts of Nerium and Argemone were
tested for their repellent effect on H. armigera. Savnur (1950) mentioned that the preparation of
good quality of medicament (bheshaja) should be a mixture of polyherbs suitable for specific
diseases in multiple forms like juice, decoction, and paste and should have potency. A repellent
activity of relative species of Cleome is also reported on spider mites (Nyalala and Grout, 2007).
Nashwa et al. (2012), studied the antimicrobial activity of six plant extracts from
Ocimum basilicum (Sweat Basil), Azadirachta indica (Neem), Eucalyptus chamadulonsis
Eucalyptus), Datura stramonium (Jimsonweed), Nerium oleander (Oleander), and Allium
sativum(Garlic) was tested for controlling Alternaria solani in vitro. In vitro study, the leaf
extracts of D. stramonium, A. indica, and A. sativum at 5% concentration caused the highest
reduction of mycelial growth of A. solani (44.4, 43.3 and 42.2%, respectively), while O.
basilicum at 1% and 5% concentration and N. oleander at 5% concentration caused the lowest
inhibition of mycelial growth of the pathogen.
Purple blotch of onion caused by Alternaria porri (Ellis) Cif. is an economically
important disease posing threat to onion cultivation in warm and humid regions. Fresh aqueous
extract of garlic (20%) was effective in causing 100 per cent inhibition of mycelial growth.
Neem oil and pongamia oil (20%) caused 76.94 and 69.94 per cent inhibition. Among the
biocontrol agents evaluated, maximum inhibition (79.5%) was recorded in T. harzianum.
Another study by Kumar (2012), that Azadirachta indica caused 66.5 % inhibition of
fungal growth of Stemphylium botryosum causing blight in garlic in vitro.
Ewekeye (2014) investigated biocontrol potential of Trichoderma harzianum and
Trichoderma koningii isolated from the the rhizosphere of the diseased tomato plants against
fungi associated with foliar diseases of tomato. The mode of antagonism was found to be by
competition for space, antibiosis and mycoparasitism.
Different Trichoderma species have good antagonistic effect on the Mycelial Growth
(MG) and Conidial Germination (CG) of A. porri (Ahmed and Lee, 2008). Bacilllus species were
more effective to inhibit the growth of Aspergillus niger, A. fumigatus, Fusarium moniliforme
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and Rhizoctonia sp. and E. coli. Inhibition of the pathogenic microorganisms was probably due
to the production of organic acids and bacteriocins (Agarry, et al., 2005).
In vitro studies by Nashwa et al. (2012), leaf extracts of Datura stramonium, Azadirachta
indica and Allium sativum at 5% caused the highest reduction of mycelia growth of Alternaria
solani at 44.4%, 43.3% and 42.2% respectively while O. basilicum at 1% and 5% concentration.
In greenhouse condition, highest reduction of disease by A. solani was achieved by extracts of
A. sativum at 5% . Under field condition, D. stramonium and A. sativum at 5% increased fruit
yield by 76.2% and 66.7% compared to the infected control.
According to Balba-Pena et al., (2006), concentrations of 10 and 15% of non-autoclaved
turmeric extracts inhibited mycelia growth by 38% and 23.2% respectively and fungal
sporulation by 71.7% and 87% respectively. When turmeric extracts were autoclaved, no
mycelia or spore germination was inhibited.
Studies by Mishra et al. (2012), where they evaluated 8 plant extracts, bio agents and
fungicides in vitro condition against Alternaria porri and Stemphylium. vesicarium causing
purple blotch and stemphylium blight of onion. Allium sativum at 10% resulted in maximum
inhibition of growth (58.05% and 57.31%respectively, followed by Aloe vera at 10 %( 53, 5%
and 47.15%).Among the bioagents, Trichoderma viride was effective in inhibition of growth (53
and 56.15%).
Goat manure had significant suppressive effects on root infection when used singly, but
the effects were less than those obtained from a mixture of goat/farm yard manure. (Alakonya et
al., 2003). According to Elizabeth Stell, author of “Secrets to Great Soil,” goat manure possesses
more organic matter--but fewer weed seeds—than most other types of animal manure, which
make it an attractive compost choice for many gardeners.
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METHODOLOGY
Materials:
Plant: Yellow ginger rhizome,
Betle leaves, Snake weed, Banana
bush, Lantana camara, Mexican
poppy, Cleome, Lagundi,
Oregano, Garlic, Aloe vera, Basil
Microbials: Bacillus sp.,
Trichoderma sp.
Test Organisms
\
Mass Culture Mass rearing in potted Mass rearing in
Plants artificial diet
MATERIALS
Disease
Organisms
Purple blotch of
garlic (A. porri)
Cercospora leaf
spot of garlic (C.
duddiae)
Early Blight
of tomato
(A. solani)
Insect Pests
Mites, thrips and
epilachna
(Garlic, pepper) Tomato (Tomato
fruitworm)
Spray Method
Dipping Method Diet Incorporation
Topical Method
Mortality
Chemosterility effect
Antifeedant/repellant effect
In vitro-agar well
diffusion method
Degree of zone of inhibition
BIOASSAY
DATA TO BE
Crude
Extract
PRODUCT
Bioinsecticide Biofungicide
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MATERIALS AND METHODS
Choice of plant materials with pesticidal potential
Some plants with known pesticidal properties were selected from secondary data i.e.,
reports in literature or from some farmers using plants as pesticide. Their distribution, abundance
and ease of mass production were also considered. The potential plant materials were planted in
herbal garden as a source of raw material for availability during mass production.
The different plants screened were: Snake weed (Euphorbia hirta), Yellow ginger
(Curcuma longa), Hot pepper (Capsicum frutescens), Betle (Piper betle), Banana bush
(Tabernaemontana pandacaqui), Kakawate leaves (Gliricidia sepium) , Lagundi leaves (Vitex
negundo), Marigold roots (Tagetes erecta), Mexican Poppy (Argemone mexicana), Lemon grass
(Cymbopogon citratus), Papaya leaves, (Carica papaya), Adelfa (Nerium indicum), Lantana
(Lantana camara), Sticky spider-flower (Cleome viscose), Acapulco (Cassia alata), Sambong
(Blumea balsamifera), Ipil-ipil (Leucaena glauca), Horse Radish (Moringa oleifera), Guava
(Psidium guajava), Aloe vera (Aloe barbadensis), Soursop (Annona muricata) Oregano
(Origanum vulgare), Onion (Allium cepa), Garlic (Allium sativum), Neem tree leaves
(Azadirachta indica) Tibak leaves, garlic waste and bamboo distillate.
Preparation and extraction methods for producing plant pesticides
Initially water, wine, vinegar and goat manure tea were used and evaluated as extractants
for the different plant materials. However plant materials extracted with water was easily
spoiled and not compatible to some plant extract causing unfavorable odor, while wine and
vinegar caused leaf burning. The use of goat manure tea (GMT) served as a carrier and prevented
the spoilage of the product. Plant extract recovery of plant materials was also noted as basis of
the active ingredient plus addition of other components to enhance toxicity, longevity etc.
Insecticidal and fungicidal property of the plant extracts
Preliminary screening was done through bioassay using the different plant extracts.
1. Mass rearing of test insects was done in order to have uniform age of sample larvae.
Choice test was done using leaf disc method and artificial diet in tomato, Helicoverpa armigera
and eggplant, Epilachna beetles. The leaf was sprayed with the plant extracts, at different levels
of concentration like pure, 75, 50, and 25 % respectively and then, air dried and wrapped with
moist cotton to maintain moisture. They were placed in a container lined with filter paper with
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the untreated leaf. Sample larva was starved for 6- 8 hours, placed in the middle of the container
and was allowed to feed for their choices. Observation for their antifeedant activity and percent
mortality was observed within 24-72 hours. The same set up was also done for insects fed with
treated artificial diet. Growth and development as affected by the different extracts were also
observed.
2. Isolation, identification of the fungal pathogens and antagonist
a. Tomato leaves showing early blight disease and garlic leaves showing typical symptoms
of purple blotch (Plate 1) were collected on naturally infected leaves from the field.
Standard isolation was followed to obtain pure cultures of the plant pathogens. Isolated
pathogens were re-cultured to obtain pure culture.
b. Antagonist; B. subtilis was taken from the Crop Protection Laboratory of the College of
Agriculture while Trichoderma sp. was isolated from bat manure tea and Bacillus sp.
from goat manure tea.
c. Pathogenecity tests and identification of the pathogen.
Tomato seedlings (var. Rosanna) were planted in pots and pathogenecity testing was
carried out by inoculating with spore suspension containing 5x106
spores /ml on foliage of 30
day old tomato plant. Symptoms of leaf spots were isolated and pure cultures were made from
the artificially infected leaves and were compared from the original culture.
Garlic (var. Ilocos White) was raised in earthen pots. After one month, plants were
thoroughly sprayed with distilled water and later covered with polyethylene bags for 24 hours.
Inoculum of 10 day old culture of A. porri (5x106) was prepared with distilled water and
separately sprayed on test plants.
Fig 1. Mass rearing of test insects, Tomato fruitworm (Helicoverpa
armigera) and preparation of plant extracts for bioassay
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Seedlings were covered and incubated for 5 days to ensure successful penetration of the
pathogen into the plant tissue. After appearance of the symptoms re-isolation was made from
diseased tissues of the artificially infected plants. Microscopic mounts were made and spores
were seen through the microscope. Identification was based on the morphological characteristics
as described by (Ellis, 1971).
Plate 1. Early blight symptoms on (a) tomato leaves and (b) purple blotch on garlic leaves
3. In vitro evaluation of botanicals against A. solani, A. porri. and C. duddiae.
Plants with known fungicidal properties were selected and used in the evaluation against
the target diseases. The agar- well diffusion method was followed to screen the efficacy of
botanicals in the laboratory against A. solani , A. porri and C. duddiae using distilled water,
Iloco vinegar and Iloco wine as extractant at concentrations of 1:1.
Bioassay tests of the different pathogens were separately conducted. About 15 ml was
poured to each of the 90mm petri dish and sterilized. Before PDA solidifies, .2 ml inoculums of
the 10 day old pathogen and .1 ml streptomycin sulfate (1:100) was incorporated into the media
to avoid bacterial contamination and allowed to solidify. Three sample wells using a .7 mm
diameter of sterile cork borer were made at 30 cm distances within the petri dish containing the
media with the pathogen. The plant extract of .1 ml was poured into each well. The plates were
incubated at room temperature for 3-5 days. PDA without extract served as control. Presence and
absence of antifungal activity was observed.
The clear zone surrounding each well indicates inhibition activity of the extract. Each
well with a clear zone were recorded using a ruler along 2 cross lines and mean inhibition was
taken in three replicates. The trial was replicated 3 times. Wider zones of inhibition were
selected in the product formulation.
a b
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4. In vitro evaluation of bio-agents against A. solani , A. porri and C. duddiae
One fungal isolate and two bacterial isolates were tested for their activity towards A.
solani, A. porri and C. duddiae. These were Trichoderma sp (isolated from bat manure tea).,
Bacillus sp. (isolated from goat manure tea ) and Bacillus subtilis taken from the College of
Agriculture, MMSU.
Mycelial discs of Trichoderma sp. was cut from actively growing colony of the fungus
and the pathogen was placed on the periphery of the petri dish at opposite sides about 3 cm. apart
from the center.
In case of the bacterial antagonists, the bacteria was streaked at the center of the petri
plate containing the medium and 2 mycelial discs of the pathogen were separately placed on the
opposite sides of the petri dish. Petri dish containing the PDA inoculated with the pathogen
served as control. After incubation, the colony diameter of the antagonists and that of the
pathogen were measured in each treatment. The percent inhibition of the pathogen over the
control was calculated following the formula:
R1 – R2
Percent Inhibition of Radial Growth (PIRG) = ----------- x 100
R1
where, R1 is the colony growth of pathogen alone; R2 ( colony growth of pathogen with
antagonists.).According to Soytong’s scale, 50% and below (low antagonistic activity), 51-60%
(mod. antagonistic activity), 61-75 %( high antagonistic activity), and more than 75% PIRG
very high antagonistic activity.
5. Product formulation
According to Oparaeke et al. (2005), the mixture of
plant extract could be use to enhance the toxicity and
activity of the products, plants with positive response
during preliminary screening were combined, formulated
and reformulated at different concentrations to became a
biopesticide products. Among the 26 plants screened plus
garlic waste, 12 plants have antifeedancy, one had mortality effect using pure extract and 16 has
antifungal activities. Initially, eight product formulation were screened as bioinsecticide and 6
Fig.3. Different product
formulation
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biofungicide plus 1 antagonist isolated against the target pest. The different biopesticide products
were also formulated into powdered and liquid form. Potential products were the following and
were tested against the target pest like tomato fruit worm in tomato, epilachna beetle in eggplant,
thrips, mites for garlic and pepper, early blight, purple blotch and cercospera leaf spots.
Bio Insecticides against the target insect pest
Product 1 (MMSU Bio-In 1) - mixture of A. mexicana, N. indicum, C. longa, E. hirta, V.
negundo + Goat Manure Tea (GMT)
Product 2 (MMSU Bio-In 2) - mixture of P. betle, C. frutescens, G. sepium, L.camara, T.
pandacaqui + GMT
Product 3 (MMSU Bio-In 3) - mixture of C. viscosa, A.mexicana, E. hirta, T. pandacaqui
+ GMT
Product 4 (MMSU Bio-In 4) - mixture of N. indicum, T. erecta, L. camara, Ocimum sp. P.
betle
Product 5 (MMSU Bio-In 5) - mixture of E. hirta, C. longa, V. negundo, C. citratus, C.
frutescens
Product 6 (MMSU Bio-In 6) - mixture of E. hirta, C. longa, P. betle, T. pandacaqui,
Ocimum sp.
Product 7 (MMSU Bio-In 7) - mixture of garlic waste and C. longa
Product 8 (MMSU Bio-In 8) - mixture of garlic waste and Neem tree leaves
Biofungicides againts early blight of tomato:
Product 1 (MMSU DF2Tm) - mixture of C. longa, A, barbadensis
Product 2 (MMSU DF3Tm) - mixture of A. sativum A. barbadensis and C. alata
Product 3 (MMSU DF4Tm) - mixture of C. longa, A. barbadensis and A. sativum
Biofungicides against purple blotch and cercospora leaf spot on garlic:
Product 1 (MMSU DF2Ga)—mixture of C. longa, O. vulgare and L. camara
Product 2 (MMSU DF3Ga)—mixture of C. longa, A. barbadensis and C. alata
Product 3 (1MMSU DF4Ga)—mixture of O. vulgare, C. longa and A. barbadensis
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Potential fungal antagonist against A. solani, A. porri and C. duddiae
(MMSU Tricho-guard) – liquid based formulation containing Trichoderma koningii
spores.
Product formulation was done and out of the 14 formulations, four was potential as
bioinsecticides, 2 as biofungicide and 1 antagonist against the target insect pest which are the
following: MMSU Bio-In 3, MMSU Bio-In 6, MMSU Bio-In 7, MMSU Bio-In 8, MMSU
DF4Tm, MMSU DF4Ga and MMSU Tricho-guard.
6. Phytotoxicity test of the formulations using wine and vinegar as extractants was
determined on tomato, pepper, eggplant and garlic using the following rating scale index (FPA
standard) after 8 hours of spraying; 1 – no crop injury, 3 – 1%-10% crop injury, 5 – 11%-20%
crop injury, 7 – 21%-30% crop injury, 9 – more than 30% crop injury.
7. Microbial load analysis and detection of pathogens present in goat manure tea.
Microbial load analysis was undertaken at the Microbiology Laboratory, Department of
Biology, MMSU, Batac City. Two samples were analyzed replicated three times. Microbials
were present in the GM tea as manifested in the samples. (See result Appendix Table 1)
8. Shelf life studies – Product formulations were evaluated and stored at room temperature.
Observations were made as to the shelf-life, change of color, presence of bubbles, foul odor and
effectiveness after months of storage.
9. Screen house experiment of the formulated products
Promising biopesticide products and concentration were also evaluated under
screenhouse condition for their products efficiency and phytoxicity. Tomato, pepper and garlic
were planted separately in pots for evaluating the formulated product’s efficacy against the target
pest and phytotoxicity. This was done by counting the initial population of the pests on sample
plants before spraying the formulated products usually 30 days after planting.
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Promising biofungicide products and concentration in vitro tests were performed as foliar
application in the greenhouse grown tomato plants. Seeds of tomato (L. esculentum), MMSU Tm
1 were planted in micro plots, .4 meter plant distances and 1 meter between rows. Formulated
plant extracts, coded as: MMSU DF2Tm, MMSU DF3Tm, MMSU DF4Tm, and one formulated
fungal antagonist and Mancozeb was used as standard fungicide. Control was sprayed with water
at the same intervals used in the other treatments.
Inoculation with A. solani suspension at concentration level of 5x106 spores ml
-1 was
sprayed two weeks after transplanting on tomato plants which was done late in the afternoon.
Treatment applications were done 2 weeks after artificial inoculation and weekly thereafter for 4
sprayings. Plants were assessed with the appearance of symptoms and the intensity of the disease
was recorded.
10. Field screening of the formulated products
Among the 14 products formulated and evaluated under laboratory and screenhouse
condition, four effective products were selected as bioinsecticides coded as MMSU Bio-In 3,
MMSU Bio-In 6 and MMSU Bio-In 7 and 2 biofungicide and 1 antagonist against the target
pest. Further screening of the different products was done under field conditions using RCBD
and replicated three times at different concentrations. The best concentration and formulation
were further verified on station and on farm with the following treatments:
Bio Insecticides:
Tomato: a) MMSU Bio-In 3, b) MMSU Bio-In 3WP50, c) Halt BT (Positive control), d)
MMSU Bio-In 8 and e) Negative control
Garlic: a) MMSU Bio-In 6 liquid and powdered form, b) MMSU Bio-In 7WP50, c) MMSU
Bio-In 8 d) Carbaryl (positive control) and d) negative control
Pepper: a) MMSU Bio-In 6 liquid and powdered form, b) MMSU Bio-In 7WP50, c) MMSU
Bio-In 8; d) Carbaryl, positive control and e) Control
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Eggplant: a) MMSU Bio-In 6WP50, b) MMSU Bio-In 7WP50, c) Carbaryl, d) MMSU Bio-
In 8 e) Control
Organic Fertilizer rate: Tomato and Garlic- 5tha-1
, Finger pepper and eggplant- 7.5tha-1
,
2 split application
Data gathering: Monitoring of pest was done starting two weeks after transplanting
except for garlic which is 30 days after transplanting. Initial counts of pest observed were
recorded before application of treatments. Based from actual observation in the field a damage
rating scale was also develop. (Appendix Rating Scale Guide Table 2-5)
Spraying was done weekly starting and as the need arises during the fruiting stage
depending upon the insect population and damage rating.
Yield data were also taken into consideration.
Bio Fungicides:
The best concentration of the formulated biofungicides were also verified on station and
on farm, specifically on the management of A. solani causing early blight of tomato, A. porri
causing purple blotch of garlic and C. duddiae causing cercospora leaf spot of garlic on station
during early planting (October) and (November) late planting. During the process of
reformulation and improvement of products another product was made (antagonist) coded as
Tricho-guard. The experiment was laid out using RCBD, 3 replications with the following
treatments:
Tomato: 1) MMSU DF2Tm, 2) MMSU DF3Tm, 3) MMSU DF4Tm, 4) MMSU Tricho-
guard, 5) Mancozeb, and 6) Control
Garlic: 1) MMSU DF2Ga, 2) MMSU DF3Ga, 3) MMSU DF4Ga, 4) MMSU Tricho-guard,
5) Mancozeb and 6) Control
On Farm
Potential bio-fungicide products found effective from the on-station trials using the
recommended organic fertilizer were further evaluated on farmers field in two sites (Batac and
Paoay) following organic and farmers’ practice. (Calendar spraying)
For tomato (MMSU Tomato 1), organic fertilizer rate was taken from the project
“Development of Organic Fertilizers and Microbials of Improved Soil Fertility in Region 1”
coded as MMSU Soil Booster following the recommended rate was 5tha- applied in two
16
applications as basal and at 25- 30 DAP. Mulch was provided (early planting) and trellised to
avoid splashing of inoculum from soil into the lower leaves and provide better air circulation.
For garlic, all the recommended organic fertilizer was broadcasted at transplanting time.
Watering was done preferably in the morning only so that leaves will be dry at night time. Goat
manure tea and FAA was applied alternately with the biofungicides as food supplement.
A total of 4 sprays were given during the cropping season. The first spray was done at
30DAT, and at 5-7 days interval. The different crops were harvested at maturity using the
maturity indices. Observations on percent disease index (PDI) were recorded at different
intervals and finally at physiological maturity of the randomly selected tagged sample plants.
All the leaves of the selected plants were scored individually. Disease rating scale used for early
blight of tomato and purple blotch of garlic was assessed following 0-5 scale (Mayee and Datar,
1986),and visual observation in the field (Cocson 2014)
Percent disease index (PDI) was computed using the formula of Wheeler (1969).
Sum of numerical ratings
PDI (%) = ______________________________ x100
Total no. of leaves x Maximum grade
examined value
Yield and yield characters like bulb weight, bulb size was also taken into consideration.
Data gathered were statistically analyzed using IRRISTAT and RCROPSTAT.
Effectiveness of the formulated products On Station were also evaluated in two
barangays at Batac City planting vegetables in comparison with farmers practice of chemical
spray. Spraying was done starting from 30 DAP and and by weekly depending upon the
population of pest applied with formulated bio-pesticide and weekly spraying of chemical.
Monitoring of pest was done weekly through visual counts and actual observation in the field
using the different rating scale guide. (See appendix table)
17
RESEARCH HIGHLIGHTS
A. Pesticidal effect of the formulated products (Laboratory and Screenhouse)
BIOINSECTICIDES
Tomato
Table 1 shows the effect of different extracts against H. armigera through choice test
using leaf disc method. Among the 16 plant extracts screened, only twelve had antifeedancy and
one had mortality effect by using pure extracts. On the other hand, the plant extracts that showed
negative pesticidal action were T. erecta, C. citratus and bamboo distillate. Notable effect was
mortality made more evident when the pure extracts were added with 20% goat manure tea
(GMT) but not at 60% concentration.
Table 1. Plant extracts with insecticidal properties against H. armigera using leaf disc method
Extracts
Pure extract 80% extract + 20%
GMT
60% extract +
40%GMT
Anti-
feedant
Mortality Anti-
feedant
Mortality Anti-
feedant
Mortality
E. hirta + - + + + +
C. longa + - + + + +
C. frustescens + - + + + +
P. betle + + + + + -
T. pandacaqui + - + + + -
G. sepium + - + - + -
V. negundo + - + + + -
T. erecta - - + -
A. Mexicana - - + +
C. citratus - - - +
C. papaya + - - +
N. indicum + - + +
L. camara + - + +
C. viscose + -
Tibak leaves + - + + + -
Bamboo distillate - - - - - -
Control
+ no feeding on treated leaf/diet or feeding damage of larva on treated leaf is ≤5% of leaf area
- mortality is below 50%
18
Knowing the stages of growth and development of a certain pest is an important
consideration for an effective pest management strategy. Uniform age (3 days old) second instar
larvae of tomato fruitworm fed with artificial diet treated with the different extracts showed
diffences in their growth stages and development. Larval developmental period of the treated
larvae ranged from 14.3-23.5 days while the control average to 18.67 days. Larvae treated with
C. viscosa did not reach the pupal stage. In addition, larvae treated with V. negundo and oregano
leaves (Coleus) died during the pupal stage. During the adult stage, longevity of the adult ranged
from one to seven days
Table 2. Effect of the different extracts on the growth and development of tomato fruitworm
using artificial diet
Plant Extracts Number of Days
Larval Period Pupa Adult
V. negundo 80%+GMT 23.5 11.0 3
C. longa 20.3 12.0 2
C. frutescens 80% +GMT 20.0 9.0 5
P. betle 19.8 8.3 7
G. sepium 19.3 12.3 3
T. erecta leaves 23.3 11.0 2
C. papaya 18.0 11.0 3
C. citratus 80%+ GMT 19.3 11.0 5
T. erecta roots 14.5 10.5 *
Coleus aromaticus 17.0 died *
C. aromaticus 80%+ GMT 20.0 died *
A.mexicana 15.0 7.7 2
L. camara 16.0 7.8 3
E. hirta 14.3 10.0
C.viscosa 80% +GMT 18.0 0.0
T. Pandacaqui 16.3 11.0 1
N. indicum 16.6 10.0 2
Lagundi tablet+GM 18.5 died *
Lagundi tablet+H2O 9.5 died *
Bamboo distillate 24.0 6.0 1
Control 18.7 12.7 4
*still pupa
19
Results of the antifeedant activity of the three formulations against H. armigera, are
shown in Table 3. All the formulations exhibited antifeedant activity but differed in effectivity
as to level of concentration. Among the three formulations, Formula 3 had the lowest percentage
surviving larvae after treatment application viz-a-viz untreated leaves from pure up to 25% level
of concentration. Formula 1 and 2 of pure extracts up to 50% concentration had lower percentage
of larvae left on treated leaf except in Formula 2, wherein the percentage of larvae left was
higher than those treated as pure extract concentration sprayed at 16-24 hours of infestation.
However, as the larvae were exposed at a longer period to the treated leaves up to 72 hours, the
percentage of larvae on treated leaves decreased. In terms of effectiveness, Formula 3 was
observed to be the most effective as compared to Formula 1 and Formula 2 because percentage
of larvae on the untreated leaves is higher than those treated with 50% concentration of extracts
to 25% concentration; this was specifically true in those treated after 24hours. For Formula 1 and
Formula 2, the most effective level of concentration was 50% to 75% plus GMT. Generally, 50%
to 75% level of concentrations plus GM are the best treatments in all the formulations. However,
formula 3 was most effective among the three formulations, although further analysis showed
that there were no significant differences among them.
Feeding damage was also taken among the different treatments using a rating scale guide
for tomato fruitworm. After 16 hours of treatment, degree of damage was almost the same
between treated and untreated leaves, that is, low to moderate damage specifically from pure to
50% concentration. However, as the larvae were exposed longer, damage rating was higher on
the untreated plants versus the treated plants. Control plants have the highest degree of damage
with a rating scale of very high to severe damage.
20
Table 3. Leaf damage by tomato fruitworm larva, Helicoverpa armigera, treated with three different formulations and level of concentrations.
Means mark with the same letter are not significantly different with each other using DMRT at 1% and 5% level
**-highly significant
*-significant Rating Scale Guide for Tomato Fruitworm
Scale Description Level
1 No damage Clear
2 ≥5% of the total leaf area eaten up Very low
3 5-10% of total leaf area eaten up Low
4 11-25% of total leaf area eaten up Moderate
5 26-50% of total leaf area eaten up High
6 51-75% of total leaf area eaten up Very High
7 76-100% of total leaf area eaten up Severe
Treatment
16 hours ** 24 hours * 48 hours ** 72 hours ns
Treated Untreated Treated Untreated Treated Untreated Treated Untreated
%
larva
Rating %
larva
Rating %
larva
Rating %
larva
Rating %
larva
Rating %
larva
Rating %
larva
Rating %
larva
Rating
Formula 1
Pure 46.7b 3 53.3b 3 36.7b 4 40.0b 3 30.0a 5 46.7a 5 23.3b 5 40.0ab 5
75% 40.0b 4 50.0b 4 26.7b 4 50.0b 4 30.0a 6 40.0ab 5 26.7b 6 40.0ab 5
50% 46.7b 4 23.3c 3 30.0b 4 43.3b 6 26.7a 6 46.7a 5 23.3b 6 46.7ab 6
25% 40.0b 5 40.0bc 4 43.3b 5 33.3c 5 36.7a 5 23.3bc 5 20.0b 5 23.3b 6
Control 73.3a 6 73.3a 6 70.0a 6 70.0a 5 46.7a 7 46.7a 7 53.3a 7 53.3a 7
Formula 2
Pure 43.3b 3 40.0b 4 46.7b 4 33.3c 4 30.0bc 4 46.7c 7 23.3b 4 46.7a 7
75% 36.7b 4 56.7b 5 30.0bc 4 56.7b 5 27.3c 4 66.7ab 7 23.3b 6 26.7a 6
50% 53.3b 4 36.7b 4 33.3bc 4 50.0b 5 40.0b 6 40.0bc 6 33.3b 7 46.7a 6
25% 56.7b 5 36.7b 5 50.0ab 5 36.7bc 5 43.3b 5 50.0bc 6 36.7b 6 40.0a 6
Control 93.3a 6 93.3a 6 70.0a 5 70.0a 5 73.3a 7 73.3a 7 50.0a 7 50.0a 7
Formula 3
Pure 20.0c 2 73.3ab 4 23.3bc 2 50.0b 5 23.3b 5 56.7a 5 23.3bc 4 46.7b 7
75% 33.3bc 2 66.7b 3 43.3b 3 50.0b 4 26.7b 5 46.7a 5 13.3c 5 43.3b 7
50% 46.7b 4 50.0b 4 36.7b 5 56.7b 5 30.0b 6 43.3a 6 23.3bc 7 46.7b 7
25% 46.7b 4 53.3b 4 30.0b 5 56.7b 5 26.7b 7 46.7a 7 40.0b 7 46.7b 7
Control 93.3a 5 93.3a 5 90.0a 5 90.0a 5 66.7a 7 66.7a 7 90.0a 7 90.0a 7
CV% 26 30 33 31
21
Eggplant
The coded formulated products (Table 4) generally reduced leaf damage caused by
Epilachna beetles on eggplant at 24 and 48 hours after treatment. Among the different
biopesticide , MMSU Bio-In 1W and MMSU Bio-In 6W had higher efficacy of control over
MMSU Bio-In 1GMT. However, result of phytoxicity test (Table 5) shows that wine caused leaf
burning of the plants in all concentrations thereby GMT was added as component of the product
coded MMSU Bio in 6.
Table 4. Leaf damage caused by Epilachna beetles on eggplant treated with three different
product formulations and at varying levels of concentrations.
Treatment/Concentrations (%)
% leaf damage
24 hours 48 hours
Treated Untreated Treated Untreated
MMSU Bio-In 1GMT
75 60.5 81.0 99.0 91.0
50 71.0 50.5 99.0 99.0
25 11.0 91.0 80.0 99.0
MMSU Bio-In 1W
75 0.0 81.0 5.5 99.0
50 25.0 94.0 31.0 99.0
25 41.0 50.0 99.0 71.0
MMSU Bio-In 6W
75 21.0 80.0 31.0 99.0
50 11.0 91.0 51.0 95.0
25 90.0 5.0 99.0 99.0
Control 97.5 99.0
Garlic
For the first set up, only seven (7) plants were screened against thrips and mites which
cause garlic tangle top (Table 5) during regular planting. These were E. hirta, C. longa, C.
frutescens, P. betle, G. sepium, along with Tibak leaves and bamboo distillate. Among the plants,
five were found to be positive in preventing tangle top to progress. For the late season planting,
additional plants were screened: V. negundo, C.viscosa, T. erecta leaf and roots, C. citratus, C.
papaya, N. indicum, L. camara and Goat manure tea (25%). Two different concentrations were
22
made using goat manure tea as solvent. G. sepium, T. erecta leaves, A. mexicana, C. viscosa and
goat manure alone showed negative effects against the test insect pests while the rest gave
positive results. Moreover, T. erecta roots showed positive result as compared to T. erecta leaves
alone which showed negative effect against the target pest. For thrips, damage was not observed
during regular planting. However, during late plantings, moderate to severe damage was
observed. Garlic is a photoperiodic plant and a good yield can be obtained during regular
planting. Observable was the great damage by thrips and mites during late season planting. In
view of this the potential pesticides during the preliminary screening for late season planting was
further investigated for during regular cropping season.
Table 5. Formulated extracts against thrips and mites causing tangle top of garlic
Extracts
Regular
Planting Late Planting
Formulated
extracts(1:2)
80% formulated
extract + 20%
(25% FGM)
50% formulated
extract + 50% (25%
FGM)
E. hirta + + +
C. longa + + +
C. frutescens + + +
P. betle + + +
G. sepium - - -
V. negundo + +
C. viscose - -
T. leaves - -
C. citratus + +
C. papaya + +
N. indicum + +
T.erecta roots + +
L. camara + +
A. Mexicana - -
Tibak +
Garlic waste +
Goat manure tea (25%) alone - -
Control
+ = 50% of the plant population did not progress/continue to show symptoms of tangle top and
show recovery
23
Sweet and finger pepper
Among the plants, four prevented leaf curling (Table 6). Plants with positive effects were
E. hirta, C. longa, P. betle and T. pandacaqui. Other plants and plant extracts combination tested
in tomato and garlic were tried in sweet pepper.
Table 6. Plant extracts against curling due to thrips, mites, and aphids.
Extracts Pure extracts (1:2)
E. hirta +
C. longa +
C. frutescens -
P. betle +
T pandacaqui +
Tibak -
Bamboo distillate -
Control -
+ = 50% of the plant population did not progress and show recovery from curling of leaves
Phytotoxicity
Table 7 shows the phytotoxicity effect of wine vinegar as extractant and goat manure tea
as solvent at different concentrations. Different concentrations with GMT were not phytotoxic to
the test crops, namely, garlic, pepper, tomato and eggplant. Similar result was also observed in
product formulations with 10% concentration of vinegar only as extractant; all the rest were
phytotoxic. Wine as extractant was phytotoxic to all the leaves of the plants regardless of the
concentration.
24
Table 7. Phytotoxicity of different formulations using wine and vinegar as extractant and goat
manure tea as solvent on selected vegetable crops
TOMATO EGGPLANT PEPPER
Formula/
Concentration
GM
tea
Wine Vinegar GM
Tea
Wine Vinegar GM
tea
Wine Vinegar
MMSU Bio-In 1
10% 1 9 1 1 3 1 1 1 1
20% 1 9 3 1 3 1 1 1 3
25% 1 9 9 1 3 3 1 3 3
50% 1 9 9 1 9 9 1 3 3
75% 1 9 9 1 9 9 1 5 3
MMSU Bio-In 2
10% 1 5 1 1 1 1 1 1 1
20% 1 5 3 1 5 3 1 3 1
25% 1 9 9 1 3 3 1 3 1
50% 3 9 9 1 9 9 1 3 1
75% 5 9 9 1 9 9 1 5 3
MMSU Bio-In 3
10% 1 3 1 1 1 1 1 1 1
20% 1 3 3 1 1 9 1 1 1
25% 1 3 3 1 1 9 1 1 3
50% 1 9 9 1 3 9 1 1 3
75% 1 9 9 1 9 9 1 1 3
MMSU Bio-In 4
10% 1 9 3 1 5 1 1 1 1
20% 1 9 3 1 9 3 1 1 1
25% 1 9 9 1 9 9 1 1 1
50% 1 9 9 1 9 9 1 3 3
75% 1 9 9 1 9 9 1 3 3
MMSU Bio-In 5
10% 1 9 1 5 1 1 1 1
20% 1 9 5 1 9 3 1 1 1
25% 1 9 9 1 9 3 1 3 1
50% 1 9 9 1 9 9 1 3 9
Crop injury; 3 – 1-10% crop injury; 5 – 11-20% crop injury; 7 – 21-30% crop injury; 9 - > 30%
crop injury
25
BIOFUNGICIDES
1. Isolation and identification of fungal pathogens
Results indicate that the tested isolate of A.solani causing early blight of tomato (Plate
2a), culture (Plate 2b) and conidia (Plate 2c) while purple blotch on garlic showing typical
symptom on lower leaf (Plate 3a), pure culture (Plate 3b) and conidia of A. porri (Plate 3c) . A.
solani was able to infect tomato and A. porri on garlic causing typical symptoms.
Plate 2. Early blight symptoms on tomato leaves (a) after inoculation of the pathogen, pure
culture on PDA (b) and conidia(c).
Plate 3. Purple blotch of garlic (a), pure culture (b) and conidia(c)
2. In vitro evaluation of botanicals against A. solani , A. porri. and C. duddiae.
Eighteen botanicals were evaluated for their antifungal activity using agar well diffusion
method against A. solani, A. porri and C. duddiae. (Table 8) and only garlic extracted in water
showed antifungal activity. Fresh extracts was easily contaminated during assay.
a b c
a b c
26
Table 8. Fungicidal activity of botanicals against A. solani, A. porri and C. duddiae in vitro using
vinegar, wine and water as extractant.
Common
Name
Botanical
Name
A. Solani A. Porri C.duddiae
Extractant Extractant Extractant
Iloco
Vinegar
Iloco
wine
Water Iloco
Vinegar
Iloco
wine
Water Iloco
Vinegar
Iloco
wine
Water
Guava P. guajava L. + + - - - - - + -
Papaya C.papaya L. - - - - - - + + -
Kakawate G.sepiumL. - + - + - - + - -
Acapulco C. alata L. - + - + + - + + -
Garlic A. sativumL. + + + + - + + - -
Lagundi V. negundoL. + + - + + - + - -
Sambong B.
balsamiferaL.
+ _ - + + - + + -
Lemon
Grass
C. citratus L.. + - - - - - + + -
Lantana L. camar.L. + - - + - - + + -
Ginger Z. officinaleL. + - - - + - + + -
Onion
multiplier
A. cepa L. + + + + - - + - -
Yellow
Ginger
C.longa L. + - + + - - + + -
Soursop A. muricata
L.
+ + - - + - - - -
Ipil-ipil L.glauca L. - - - - - - + - -
Horse
Radish
M. oleifera L. + - - + - - + - -
Oregano O.vulgare L. + - - + + - + + -
Aloe A.barbadensis
L.
+ + - + + + - + -
Betle P. Betle L. + - - - - - + - -
+ - presence of antifungal activity
- - no antifungal activity
27
Alternaria solani
The antifungal activity of the botanicals using vinegar and wine was shown in Table 9.
Among the botanicals, A. sativum extracted in vinegar was most effective in inhibiting mycelial
growth with mean inhibition zone of 38.00 mm, followed by A. barbadensis (30.03mm), and V.
negundo (26.67mm) and P guajava (21.67mm). The rest also showed mycelial inhibition which
range from 10.03 mm to 19.17mm mean diameter.
A. sativum extracted in wine was most effective in inhibiting fungal growths with 40.03
mm followed by P. guajava (38.00mm ), G. sepium (34.67mm), V negundo (30.03mm), A cepa
(27.50 mm), A. barbadensis (26.67 mm), A. muricata (25.17mm) and C. alata (21.67mm).
Alternaria porri
Among the botanicals, highest inhibition of mycelial growth using vinegar as extractant
were; C.. longa(15.33 mm), M. oleifera (15.03 mm),A. sativum (13.33 mm), A. cepa (13.33mm)
and V. negundo (11.03 mm). Least inhibition was observed from G. sepium, C. alata, B.
balsamifera, L. camara, O. vulgare, A. barbadensis with fungal inhibition of 10.03 mm.
Using Iloco wine as extractant, highest inhibition was observed from A. barbadensis
(16.33 mm), O. vulgare (14.17 mm), V. negundo (13.17mm) and B. balsamifera (11.00 mm).
Lesser inhibition was observed from C. alata, Z. officinale, and A. muricata with a diameter
zone of 10.00 mm.
Cercospora duddiae
Highest inhibition of fungal growth using vinegar as extractant was observed from Z.
officinale (30.50 mm), A. sativum (30.00 mm), O. vulgare (23.33 mm), B. balsamifera (21.00
mm), L. glauca (18.16 mm), V. negundo (18.00). and ,C. longa, A. cepa(13.33 mm). Lesser
diameter of inhibition was observed from C. citratus (15.66 mm), M. oleifera, (15.00 mm), C.
papaya (14.33mm), C. alata (13.66) and P. betle(12.00) least inhibition was on L. camara
(10.00mm).
Iloco wine as extractant to the botanicals also manifested inhibition of the pathogen.
Highest diameter of inhibition zones was observed from A. barbadensis (28.16mm) followed by
P. betle (23.66 mm),Z. officinale (20.50 mm),L. camara(19.83 mm), B. balsamifera (19.16
mm), and C. longa (17.66), . Least inhibition was noted from O. vulgare (14.00 mm), P.guajava
(13.50 mm),C. papaya (12.00mm), and C. citratus (11.66mm).
28
Table 9. In vitro evaluation of botanicals against A. solani, A. porri and C. duddiae using Iloco
vinegar and Iloco wine.
Botanical Name
Mean Inhibition Zone of Mycelial Growth (mm)
A. solani
A. porri C.duddiae
Extractant (1:1) Extractant(1:1) Extractant(1:1)
Vinegar Wine Vinegar Iloco wine Vinegar Wine
P. guajava L. 21.67c 38.00a - - - 13.50d
C. papaya L. - - - - 14.33cde 12.00d
G. sepium - 34.67ab 10.03 c - 10.83e -
C. alata - 21.67d 10.03.c 10.3c 13.66de 10.83d
A. sativum 38.00a 40.03a 13.33 ab - 30.00a -
V. negundo 26.67b 30.03bc 11.03 bc 13.17b 18.00bcd -
B. balsamifera 10.03f - 10.03 c 11.00c 21.00bc 19.16c
C.citratus L. 13.33ef - - - 15.66cde 11.66d
L. camara 11.33ef - 10.03c - 10.00e 19.83c
Z. officinale 18.33cd - - 10.03c 30.50a 20.50
A. cepa 11.67ef 26.50cd 13.33ab - 18.00bcd -
C. longa 15.33de - 15.33a - 18.00bcd 17.66c
A. muricata L. 15.50de 24.17cd - 10.03c - -
L. glauca - - - - 18.16bcd -
M oleifera 18.33cd - 15.03a - 15.00cde -
O. vulgare 19.17cd - 10.03 c 14.17b 23.33b 14.00d
A. barbadensis 30.03b 26.67cd 10.03c 16.33a - 28.16a
P. betle 11.67ef - - - 12.00de 23.66b
Level of
significance
** ** ** ** ** **
CV (%) 13.3 38.00a 12.4 7.0 20.4 9.9
3. In vitro evaluation of bio-agents against A. solani, A. porri and C. duddiae
All the tested antagonists inhibited the growth of the tested pathogens in vitro, Table 10
and Plate 4. The antagonist effectively showed maximum inhibition (81.20% to 83.30%) against
A. solani. High antagonistic activity was also observed against A. porri, while lower antagonistic
activity against C. duddiae.
29
Table 10. Effect of the different antagonists on the growth of A. solani.A. porri and C. duddiae.
Antagonist
A. solani A. Porri B. Duddiae
Colony
diameter
(mm)
Inhibition
(%)
Colony
diameter
(mm)
Inhibition
(%)
Colony
diameter
(mm)
Inhibition
(%)
Trichoderma sp. 15.10 81.20
(64.30)
30.0 62.50a
(52.24)
23.3
36.26b
(36.15)
B. subtilis 13.33 83.30
(65.96)
28.3 64.61b
(53.49)
23.3
37.12a
(37.12)
GM bacteria
(Bacillus sp.)
13.33 83.30
(65.960)
28.3 64.61b
(953.49)
21.6
36.10b
(35.89)
Control 80.00 0(00) 80.00 0(00) 36.6 0(00)
Level of
significance
** **
CV (%) 3.8 15.2
*Data in parenthesis are arc sine transformed values
Plate 4. In vitro preliminary evaluation of the fungal antagonist.
Plate 5.In vitro evaluation of Bacillus sp. isolated from goat manure tea
showing inhibition of test pathogen.
30
The most effective is MMSU Tricho-Guard and MMSU DF4Tm, which was comparable
with Mancozeb. These products had lesser disease intensity, more disease reduction and
eventually higher yield (Table 11). Lesser reduction of disease intensity was obtained from
MMSU DF2Tm and MMSU DF3Tm and had lower yield (8.6tha-1
-9.4tha-1
) while the control
was most infected and obtained the lowest yield (4.1tha-1
). View of the experiment is shown in
Plate 6.
Table 11. Efficacy of biofungicide products against early blight disease of tomato
plants under screenhouse condition
Treatments Disease
severity (%)
Disease
reduction
(%)
Yield (t/ha)
MMSU DF2Tm
**
52.4b
36.64
**
9.4ab
MMSU DF3Tm 52.0b 37.12 8.6b
MMSU DF4Tm 40.7c 50.74 12.3a
MMSU Tricho-guard 35.9c 56.59 11.5ab
Mancozeb 35.4c 57.68 11.3ab
Infected control 82.7a 4.1c
CV(%) 10.8 12.7
Values in the column followed by different letters indicate significant differences among
treatments according to LSD test (P=05)
Plate 6. View of the greenhouse experiment on early blight of tomato using the different
biofungicides.
Control
31
EFFECTIVENESS OF FORMULATED PRODUCTS UNDER FIELD CONDITIONS
BIOINSECTICIDES
Tomato
Formulated botanical insecticides MMSU Bio-In 3 obtained a lower damage rating scale
against tomato fruit worm as compared to control plants although insignificant differences was
noted with the other treatments in both on- station and on- farm trials (Fig. 5). The yield
followed similar trend wherein plants sprayed with botanical products obtain numerically higher
yield as compared to plants sprayed with insecticide and control plants., although it was higher
from on-farm trial (Fig. 6).
Figure 5. Damage rating scale of tomato fruitworm sprayed with botanicals planted on
farm and on station
Rating Scale Guide for Tomato Fruitworm
Scale Description Level
1 No damage Clear
2 ≥5% of the total leaf area eaten up Very low
3 5-10% of total leaf area eaten up Low
4 11-25% of total leaf area eaten up Moderate
5 26-50% of total leaf area eaten up High
6 51-75% of total leaf area eaten up Very High
7 76-100% of total leaf area eaten up Severe
32
Figure 6. Marketable yield of tomato sprayed with formulated pesticides planted on
station and on farm
Garlic
During the first trial no significant differences were obtained among the treatments in
terms of yield in garlic plants sprayed with MMSU Bio-In 6 as compared to those plants sprayed
with insecticide including the control in both trials (Fig. 7). It was also observed that the
occurrence of tangle top was negligible during the conduct of experiment on station and minimal
on farmer’s field. Instead purple blotch and cercospora leaf spot were prevalent during the later
stage of the plant. Thrips was also observed after the bulbs were already formed and the lowest
damage rating scale was observed by plants treated by Bio-In 6. (Fig. 8).
Figure 7. Yield of garlic planted at on- farm and on- station fields
33
Figure 8. Percent damage of thrips and yield of garlic sprayed with MMSU BioIn 6 and
insecticide
During the second trial, two biopesticide product coded MMSU Bio-In 7 and MMSU
Bio-In 8 was included in the evaluation in garlic as a result of product reformulation. Higher
yield was noted on plants sprayed with MMSU Bio-In8 on both trials with a yield increase of
19.4% over the control although highest yield was obtain on station trial while the yield on farm
is comparable among the treaments (Table 12). Lowest yield was obtained on plants with
MMSU Bio-In 7 in both trials. This can be attributed to the garlic waste particularly on the
leaves as one of the component of the product that might favor the development of the disease,
however this product is potential in pepper leaf curling. MMSU Bio-In 8 have also a garlic waste
component but it was combined with neem tree leaves to enhance the toxicity of the products. It
was also noted that yield of the garlic is better as compared to the first trial and this might due to
the fact related to the weather conditions tha favor growth an devlopment of bulbs,In addittion
purple blocth infestation was very low this year as compared to previous years and only
noticaeable during the later stage of the plants. The same is true with the incidence of tangle top
although higher percentage was observed in control plants in the on farm trial. Garlic productin
is a hit and miss venture, pest is always there and this should be prevented to become damaging
in order to obtained a good yield.
34
Table 12. Garlic yield and pest damage rating sprayed with differrent biopesticide
Treatment/
Biopesticide
Purple Blotch (%) Tangle Top (%) Yield (tha-1
) Yield
Increase
(%)
On
Station
On
Farm
On
Station
On
Farm
On
Station
On
Farm
MMSU Bio-In 6 16 12 8 8 4.10 3.50 2.7
MMSU Bio-In 7 20 24 8 8 2.85 3.00 -26.0
MMSU Bio-In 8 16 12 8 8 5.25 4.00 19.4
Carbaryl 16 16 16 24 4.30 3.85 9.35
Control 20 20 12 20 4.00 3.40
Finger pepper
Yield of finger pepper was higher in plants treated with MMSU Bio-In 6 and MMSU
Bio-In 7, as compared in plants applied with the chemical pesticide and the control. The same
findings were noted on leaf curling on the plants but no significant differences. (Fig.9).
Figure 9. Curling of leaves and yield of finger pepper sprayed with insecticide
Rating Scale Guide
Scale Description Level
1 No damage Clear
2 ≥5% of the total leaf area curl up/down Very low
3 5-10% of the total leaf area curl up/down Low
4 11-25% of the total leaf area curl up/down Moderate
5 26-50% of the total leaf area curl up/down High
6 51-75% of the total leaf area curl up/down Very High
7 76-100% of the total leaf area curl up/down Severe
35
During the second trial biopesticide products that was evaluated in garlic was also tried in
pepper since both crops have the same type of damage feeding (sucking type) specifically on leaf
curling due to thrips mites and aphids (Table 13). It was noted that the plants sprayed with the
three MMSU biopesticide products including the chemical insecticide have low to moderate
damage as compared to the control from moderate to high damage. Highest yield was noted in
plants treated with MMSU Bio-In 7 followed by MMSU Bio-In 6 with a percent yield increase
of 24.85 and 12.25% respectively over the control which is 4.0%.
Table 13. Pepper yield and damage rating on the curling of leaves sprayed with different
biopesticide
Treatment/
Biopesticide
Curling of Leaves Rating
Scale
Yield (tha-1
) Yield
Increas
e (%)
On Station On Farm On Station On Farm
MMSU Bio-In 6 4.40 9.2 6.0 4.00 12.25
MMSU Bio-In 7 4.10 10.8 7.0 3.50 24.85
MMSU Bio-In 8 4.25 8.0 4.0 3.50 10.00
Carbaryl 4.30 8.2 5.9 3.50 4.00
Control 5.00 7.1 5.9 4.50 0
Eggplant
Comparable yield was obtained from eggplant treated with biopesticides, although
numerically, plants treated with MMSU Bio-In 6 obtained the highest yield and the lowest was
the control plants. High damage rating for shoot/fruitborer was observed in control plants as
compared to the formulated product- and insecticide- treated plants. (Fig.10)
Figure 10. Shoot/fruit borer, Epilachna damage and yield of eggplant sprayed with insecticide
36
The same findings during the second trial (Table 14) that the MMSU formulated
botanical insecticides was comparable with the commercial insecticide in terms of damage rating
and the highest was observed in the control plants. Plants treated with MMSU Bio-In 6
consistently gave numerical higher yield in both trials. Overall the different products showed no
significant differences when tested under field conditions and comparable to chemical
insecticides, however numerically control plants obtained the lower yield as compared plant
sprayed with botanical pesticides. The different products can be useful for managing the target
pest and a suitable alternative of chemical pesticide.They can be integrated with other pest
management strategies and systems.
Table 14. Epilachna, shoot/fruit borer, leafhopper damage rating and yield of eggplant sprayed
with different biopesticide.
Treatment/
Biopesticide
Epilach Damage Shoot/Fruit
Borer
Leafhopper
Damage
Yield
(tha-1
)
Yield
Increase
(%)
On
Farm
On
Station
On
Farm
On
Station
On
Farm
On
Station
On
Station
MMSU Bio-In
6WP50 1.0 1.5 1 1 4.0 4.0 10.0 10
MMSU Bio-In
7WP50 1.5 1.5 1 2 4.1 4.0 10.0 10
MMSU Bio-In
6 1.0 1.5 1 2 4.4 3.9 12.0 8.3
MMSU Bio-In
7 1.0 3.0 3 3 4.1 4.0 10.2 7.8
Carbaryl 1.0 1.0 1 2 4.4 4.2 11.7 5.9
Control 3.0 3.0 3 5 7.6 4.5 11.0
37
BIOFUNGICIDES
Tomato (On Station)
Efficacy of botanical fungicides against early blight of tomato
The biofungicide-treated plots significantly lower disease index ranging from 49.3% to
53.3% in October planting which was comparable to the chemical fungicide (48.3 %). The
control exhibited higher disease severity with 76.0%, (Table 15).
In November planting, lesser PDI values (29.8 % -31.7%) were observed in the treated
plots but comparable to the chemical fungicide (29.3%) in comparison to that of untreated plots
(43.4%). Higher numerical values were obtained in October planting due to high relative
humidity and some rains during the growing season whereas November planting preceded drier
condition.
All the botanical products significantly lower PDI which is comparable to the chemical
fungicide. The control significantly higher PDI. PDI was lower in case of the botanical-treated
and the fungicide- treated plants in comparison to the control plants.
In terms of fruit infection in both plantings, using the MMSU biopesticide products and
chemical fungicide spray showed significantly lesser fruit infection range (1.68% to .78%) for
October planting and (1.00% to .65%) for November planting with comparable results while the
control obtained the highest number of fruits infected at 6.23% and 4.02% respectively.
No. of fruits was not affected by the different treatments in between planting while the
treated plants in November planting showed significant increase of fruits harvested compared to
the control.
Tomatoes planted in November and sprayed with botanical fungicides and the chemical
fungicides showed comparable significant yield increase from 39.95tha-1
to 41.26tha-1
.
Apparently, higher yield was attributed to bigger fruits in the November planting and
consequently heavier fruit weight. Lower yield was obtained from the control at 26.05 tha-1
.
38
Table 15. Severity of early blight caused by A. Solani on growth and yield of tomato sprayed
with different biofungicides at different planting.
Treatment PDI at 75 DAP Fruit Infection (%) No. of fruits per
plant
Fruit Yield (tha-1
)
Oct.
Planting
Nov.
Planting
Oct.
Planting
Nov.
Planting
Oct.
Planting
Nov.
Planting
Oct.
Planting
Nov.
Planting
** ** ** * ns ** ns **
Control 76.0b 43.4b 6.23b 4.02b 18.60 18.51b 23.30 26.05b
MMSU
DF2Tm
53.3a 31.4a 1.68a 1.00a 21.86 25.98a 26.71 39.95a
MMSU
DF3Tm
53.3a 32.5a 1.04a 0.75a 24.77 24.90a 29.42 39.22a
MMSU
DF4Tm
49.3a 29.8a 0.78a 0.65a 25.58 25.54a 34.07 41.26a
Mancozeb 48.3a 29.3a 0.99a 0.68a 26.40 27.62a 30.15 40.00a
CV (%) 6.8 8.6 26.8 73.1 20.0 9.3 13.1 10.6
Means in a column followed by a common letter are not significantly different at 5% level by
DMRT.
*- significant at 5% level
**- significant at 1% level
Garlic (On Station)
Efficacy of formulated products against foliar disease of garlic (on station)
Purple Blotch
In plant disease management, time of planting is a part of the strategy to manage diseases
particularly fungal diseases.
At 75 DAP, the biofungicide were at par lower in PDI with comparable result to the
chemical fungicide compared to the control in both plantings (Table 16). Generally, however,
October planting had lower infection as manifested by the numerical values while November
planting had higher numerical values. The disease increment in November planting is due to the
source of inoculum developed from infected plants in the October planting favored by the dry
humid environment. This result confirms previous studies by Pascua et. al, 2002 that October 15
planting obtained a disease rating of 2 in which 30- 50 % leaf area was infected with purple
blotch.
39
Average Bulb Weight
Heavier bulbs (9.35g to 9.98g) were observed in the biofungicide-treated plots and were
comparable to the chemical fungicide- treated plants (9.75g) in the October planting, in contrast
to the control with only 7.38g. Similar results were obtained from November planting where
bigger and heavier bulbs were obtained from using bio-fungicide which was comparable to the
chemical fungicide- treated plots.
Yield
In October planting, higher yields (2.94 to 3.10 tha-1
) were obtained in the biofungicide-
sprayed plants which was comparable to the chemically- sprayed plants (2.89 tha-1
). Lower yield
was obtained from the control with only 2.35tha-1
. The same findings during November planting
wherein the use of MMSU DF4Ga (2.63tha-1
) and MMSU DF3Ga (2.42 tha-1
) obtained
comparable yield results to the chemical fungicide- sprayed plants with 2.49 tha-1
but not
significantly different using the botanical DF2Ga with 2.19 tha-1
. Lowest yield was obtained
from the control plants with only 1.78tha-1
. Disease is a limiting factor in garlic cultivation,
where most of the photosynthetic area of the plant is destroyed which resulted to lower yield in
plants with more intense disease damage.
Though disease severity was reduced by spraying with the biofungicide formulations and
the chemical check, apparently, October planting had lower disease intensities as manifested in
the lower numerical values. On the other hand, the use of biofungicide formulations also
reduced disease intensity during November planting though disease severity was more apparent.
Generally, the biofungicide- sprayed plants including that of chemical fungicide obtained
higher and comparable yield in both plantings due to bigger bulbs which resulted in higher yield.
Table 16. Field evaluation of the biofungicide products against purple blotch of garlic
Treatment
PDI at 75 DAP Ave. bulb wt.( (g) Yield (tha-1
)
Oct. Nov. Oct. Nov. Oct. Nov.
** ** * * * *
Control 38.86a 46.26a 7.38b 5.68b 2.35b 1.78b
MMSU DF2Ga 28.04b 34.93b 9.35a 6.98ab 2.94a 2.19ab
MMSU DF3Ga 27.34b 34.50b 9.50a 7.73a 3.06a 2.42a
MMSU DF4Ga 25.96b 34.73b 9.98a 8.38a 3.10a 2.63a
Mancozeb 25.60b 32.93b 9.75a 8.23a 2.89a 2.49a
CV (%) 8.2 7.2 9.3 12.0 8.2 10.1
40
Cercospora Leaf Spot
The effect of the plant based botanical products on Cercospora leaf spot infection,
expressed in percent disease index were taken at 75 DAP is shown in Table 17.
Percent Disease Index
Disease development in October planting was only apparent at the later stage of the
plants and was quite low. However, significant difference were noted where MMSU DF4Ga
obtained lowest infection of 2.22 % which was comparable to MMSU DF3Ga (2.29 %). Higher
infections were noted on the control (2.35 %), MMSU DF2Ga (2.35 %) and the use of standard
fungicide (Mancozeb) with 2.38 %. For the November planting no significant differences in
disease severity were noted among the treatments.
Average Bulb Yield
Generally, the biofungicide treated plants obtained heavier bulbs with comparable results
from the chemical fungicide in both plantings (October and November)
The use of formulated products, namely, MMSU DF4Ga and MMSU DF2Ga obtained
higher yield of 3.13 t/ha and 2.89 t/ha respectively, chemical fungicide (3.04 t/ha) in October
planting (Table 17). Lesser yield was obtained from the control (2.43 t/ha) and MMSU DF2Ga
(2.43 t/ha).
This was also November planting wherein the formulated products obtained higher yields
(2.79 t/ha-3.03 t/ha) which was comparable to the chemical fungicide (3.00 t/ha). Lowest yield
was obtained from the control with only 2.35 t/ha.
Table 17. Field evaluation of bio fungicide products against cercospora leaf spot of garlic
Treatment PDI at 75 DAP Ave. bulb weight (g) Yield (tha-1
)
Oct. Nov. Oct. Nov. Oct. Nov.
** Ns ** ** ** **
Control 2.35a 2.48 7.90b 7.49b 2.43b 2.35b
MMSU DF2Ga 2.35a 2.37 9.23a 8.88a 2.89a 2.79a
MMSU DF3Ga 2.29ab 2.49 8.13b 8.98a 2.55b 2.83a
MMSU DF4Ga 2.22b 2.46 9.94a 9.65a 3.13a 3.03a
Mancozeb 2.38a 2.44 9.70a 9.56a 3.04a 3.00a
CV (%) 2.5 5.1 5.8 4.9 5.9 5.0
41
ANTAGONIST (MMSU Tricho-guard)
Another product was developed as antagonist (coded as MMSU Tricho-guard) as a result
of the improvement of the reformulated product against the target diseases and they were also
tested under field conditions for tomato and garlic.
For tomato, all the treatments significantly reduced early blight of tomato under field
condition (Table 18). However, greatest reduction of the disease was achieved in treatments
MMSU DF4Tm (35%), Tricho-guard (34.3%) which was comparable to standard fungicide
(Mancozeb) with 37.2%. In terms of yield, formulated products manifested higher number of
fruits per plant (16-18) compared to the control of only 8. Tomato plants sprayed with MMSU
DF4Tm obtained the highest yield of 26.1 t/ha with an increased fruit yield by 60.91% due to
bigger fruits. In contrast, MMSU DF3Tm, MMSU Tricho-guard and Mancozeb increased yield
moderately ranging from 54.5% to 57.67% compared to the control.
Table 18. Efficacy of the different improved biofungicides on early blight disease and yield of
tomato under field condition.
Treatments
Percent
Disease
index(%)
Disease
Reduction (%)
No. of fruits
/plant
Yield
(t/ha)
Yield
Increase (%)
MMSU DF2Tm
**
51.5b
20.1
**
16a
**
19.7b
48.22
MMSU DF3Tm 47.5c 26.4 16a 22.2ab 54.05
MMSU DF4Tm 41.9d 35.0 19a 26.1a 60.91
MMSU Tricho-
guard
42.4d 34.3 18a 24.1ab 57.67
Mancozeb 40.5d 37.2 17a 22.5ab 54.66
Control 64.5a 8b 10.2c
CV (%) 3.8 12.5 12.6
Values in the same column by different letters indicate significant differences among treatments
to the LSD test
For garlic, there was a disease reduction of purple blotch using treatments MMSU
DF4Ga, MMSU Tricho-guard and the standard fungicide ranging from 23.78% to 25.81%
compared to the control (Table 19). All the treatments increased yield over the control however
highest yield was obtained from plants sprayed with the bio- fungicide MMSU DF4Ga and
MMSU Tricho-guard at 4.2 tha-1
respectively which was due to bigger bulbs. The control
obtained the lowest yield of 3.4 tha-1
due to smaller bulbs and higher disease intensity.
42
Table 19. Efficacy of the improved biofungicide against purple blotch and yield of garlic
Treatments Percent
disease
Index (%)
Disease
Reduction
(%)
Ave. bulb
wt.(g)
Yield
tha-1
Yield
increase
(%)
MMSU DF2Ga
**
42.8b
13.0
*
15.8ab
*
4.1ab
17.07
MMSU DF3Ga 43.3b 11.99 15.2b 3.8ab 10.52
MMSU DF4Ga 36.5c 25.81 16.7ab 4.2a 19.04
MMSU Tricho-guard 36.8c 25.20 16.8a 4.2a 19.04
Mancozeb 37.5c 23.78 15.6ab 3.9ab 12.82
Control 49.2a 0 13.6c 3.4b 0
CV(%) 2.7 5.7 7.0
Values in the column followed by different letters indicate significant differences among
treatments according to LSD test
ON FARM TRIAL of BIOFUNGICIDE PRODUCTS
TOMATO
Tomato Percent Disease Index
Early blight symptoms were assessed 75 DAT in both sites (Table 20). The organic
practice had lower PDI in Paoay and Batac with 35.1% and 31.47 % respectively. The farmers
practice obtained higher PDI at 42.7% and 43.4%. Using the organic practice, there was a
disease reduction of 17.7% in Paoay and 27.4% in Batac site.
Yield
Marketable yield was also higher in Paoay using organic products (26.9 tha-1
) and Batac
(20.5tha-1) as compared to Farmers’ practice in Paoay obtained lower marketable yield of 22.2
t/ha and 18.5 in Batac site. Yield increase of 17.2% (Paoay) and 9.7% (Batac) was realized using
the organic technology. These results of a farm trial indicated that the application of organic
fertilizer enhances vigor and in combination with the plant –based biofungicides effectively
reduced disease severity and an increase in yield.
The benefit of incorporating organic fertilizer and biofungicides in the management of
early blight was considerable. It can be used successfully as an alternative tool for disease
management thereby reduced chemical fungicide application. Biopesticide is safe and no serious
hazard and pesticide toxication.
43
Table 20. Disease severity, marketable yield and plant vigor of tomatoes using biofungicide
products and biofertilizers in Batac and Paoay site.
Particulars
PDI at 75
DAT (%)
Disease
reduction (%) Marketable
yield (t/ha)
Yield
Increase (%)
Plant Vigor*
Paoay
Batac
Paoay
Batac
Paoay
Batac
Paoay
Batac
Paoay
Batac
Organic
practice
35.1 31.4 17.7 27.4 26.9 20.5 17.2 9.7 1 2
Farmers’ practice
42.7 43.4 22.2 18.5 3 3
*- taken 30 DAT
Vigor rating: 1= very vigorous, 2=vigorous, 3=mod. vigorous, 4= weak, 5=very weak
Plate 7. Field view of the on- farm trial (Tomato and garlic) farmers’ fields in Paoay.
Garlic
Plate 8. Field view of the on-farm trial (Tomato and garlic) in farmers’ fields of Batac
Tomato Garlic
Tomato
Farmers Practice
Organic Practice
44
GARLIC
Purple blotch of garlic
Between the two sites (Paoay and Batac), lower infection (Table 21) caused by purple
blotch was observed in organic practice (27.26%-35.7%) than the farmers practice (29.60%-
39.5%) at 75 DAP. A disease reduction of 7.9% (Paoay) and 9.6% (Batac) was observed in the
organic practice. Bulb yield was also higher in organic practice (Paoay) with 3.45 tha-1
while the
farmers practice was lower at 2.80t ha-1
. In the case of Batac site, organic practice obtained lower
yield of only 2.9 tha-1
. For the two sites there was a considerable increase in yield of 18.8 %
(Paoay) and 20.6 % (Batac) using the organic practice. Generally, the benefits of incorporating
biopesticides and the use of organic fertilizer in the management of purple blotch and
Cercospora leaf spot are considerable.
Table 21. Disease severity of purple blotch and bulb yield of garlic in organic and farmers
practice, DS 2012-2014.
Particulars
PDI of Purple blotch at
75 DAP (%)
Reduction
(%)
Bulb Yield (tha-1
) Increase
(%)
Paoay Batac Paoay Batac Paoay Batac Paoay Batac
Organic
Practice
27.26 35.7 7.9 9.62 3.45 4.8 18.8 6.25
Farmers
Practice
29.60 39.5 2.80 4.5
COST OF PRODUCING BIOPESTICIDE PRODUCTS
The cost of the produced biopesticides was based on material inputs, labor cost and
other miscellaneos costs, (Appendix Table 8). The cost of biopesticide was Php155.60,
biofungicde was Php 175.50 while the antagonist Php 93.00. The availability of the material in
the locality is a contributory factor in producing the different products.
COST AND RETURN ANALYSIS
TOMATO
Tomato treated with MMSU Bio-In 3 obtained the highest net income of Php 369,776
followed by MMSU Bio-In F3WP50, Halt, MMSU Bio-In 7 and Control. Moreover, it was
observed that it gave also lowest production cost of Php 4.33/kg.
45
Table 23. Cost and return analysis of growing tomato per hectare sprayed with different
bioinsecticides (On Station)
TREATMENT/
BIOINSECTICIDE GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET
INCOME
(per hectare)
ROI
(%)
UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio-In 3 447,250 77,474 369,776 477.3 4.33
MMSU Bio-In
F3WP50
426,250 90,039 336,211 373.4 5.28
MMSU Bio-In 7 307,250 78,028 229,222 293.8 6.35
Halt(BT) 399,000 79,637 319,363 401.0 4.99
Control 371,750 74,704 297,046 397.6 5.02
In table 24 it also shows that MMSU Bio-In 3 in wettable powder form gave the highest
net income per hectare. It was perceived that the use of bioinsecticides gave highest percentage
investment return and the lowest production cost per kg. as compared to the other treaments.
Highest production cost of Php 4.49/kg was noted in the use of chemical pesticide.
Table 24. Cost and return analysis of growing tomato per hectare sprayed with different
bioinsecticides (On Farm)
TREATMENT/
BIOINSECTICIDE
GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET INCOME
(per hectare)
ROI
(%)
UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio-In 3 503,250
77,474
425,776
549.6
3.85
MMSU Bio-In
3WP50
545,500
90,039
455,461
505.8
4.13
MMSU Bio-In 7 503,000
78,028
424,972
544.6
3.88
Halt(BT) 443,250
79,637
363,613
456.6
4.49
Control 439,000
74,704
364,296
487.7
4.25
46
GARLIC
Comparative cost and return analysis shows the profitability of growing garlic sprayed
with different bioinsecticides (On Station) (Table 25). Garlic sprayed with MMSU Bio-In 8 was
noted to have the highest return of income of Php 371,942. The ROI was at 243.0%. Lowest net
return was obtained from plants treated with MMSU Bio-In 7 with a generated income of Php
131,942 per hectare.
Table 25. Cost and return analysis of growing garlic per hectare sprayed with different
bioinsecticides (On Station) TREATMENT/
BIOINSECTICIDE GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET INCOME
(per hectare)
ROI
(%)
UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio-In 6 410,000 153,295 256,705 167.5 37.4
MMSU Bio-In 7 285,000 153,058 131,942 86.2 53.7
MMSU Bio-In 8 525,000 153,058 371,942 243.0 29.2
Carbaryl 430,000 154,667 275,333 178.0 36.0
Control 400,000 149,734 250,266 167.1 37.4
Garlic treated with MMSU Bio-In 8 obtained the highest net return per hectare of Php
246,942 and ROI of 161.3% as shown in Table 26. Additionally, it was noted that it has the
lowest production cost/kg of Php 38.26. This was followed by carbaryl with a production cost/kg
of Php 40.2 and net income per hectare of Php 230,333 and ROI of 148.9 %. The least income
was obtained from MMSU Bio-In 7 with a net income per hectare of Php 146,942 and ROI of
96.0%, thus, it has the highest production cost/kg of Php 51.02.
Table 26. Cost and return analysis of growing garlic per hectare sprayed with different
bioinsecticides (On Farm)
TREATMENT/
BIOINSECTICIDE
GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET INCOME
(per hectare)
ROI
(%)
UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio In-6 350,000 153,295 196,705 128.3 43.8
MMSU Bio In-7 300,000 153,058 146,942 96.0 51.0
MMSU Bio In-8 400,000 153,058 246,942 161.3 38.3
Carbaryl 385,000 154,667 230,333 148.9 40.2
Control 340,000 149,734 190,266 127.1 44.0
47
PEPPER
The cost and return analysis in growing pepper per hectare sprayed with different
bioinsecticides was showed in Table 27. Results revealed that the pepper treated with Bio-In 7
gave the highest net income of Php 51,972 followed by Control, Bio-In 6, Insecticide and Bio In
8. In terms of ROI, Bio in 7L gave the highest percentage of 59.0.
Table 27. Cost and return analysis of growing pepper per hectare sprayed with different
bioinsecticides (On Station)
TREATMENT/
BIOINSECTICIDE
GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET
INCOME
(per hectare)
ROI
(%)
UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio-In 6 120,000 88,266 31,734 40.0 14.7
MMSU Bio-In 7 140,000 88,028 51,972 59.0 12.6
MMSU Bio-In 8 80,000 88,028 -8,028 -9.1 22.0
Carbaryl 118,000 89,637 28,363 31.6 15.2
Control 118,000 84,704 33,296 39.3 14.4
Table 28 shows the cost and return analysis of pepper on farm. The highest gross
income, net income per hectare and ROI were obtained from treatment applied with MMSU Bio
In-7. On the other hand, the lowest gross income, production cost, profit per hectare, and ROI
were derived from no pesticide application (control).
Table 28. Cost and return analysis of growing pepper per hectare sprayed with different
bioinsecticides (On Farm)
TREATMENT/
BIOPESTICIDE
GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET INCOME
(per hectare)
ROI UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio In-6 184,000 88,266 95,734 108.5 9.6
MMSU Bio In-7 216,000 88,028 127,971 145.4 8.2
MMSU Bio In-8 160,000 88,028 71,972 81.8 11.0
Carbaryl 164,000 89,637 74,363 83.0 10.9
Control 142,000 84,704 57,296 67.6 11.9
EGGPLANT
The cost incurred and return generated in growing eggplant sprayed with different
bioinsecides was computed and presented in Table 29. Eggplant treated with MMSU Bio-In 6
gave the highest net return with a computed net income of Php 268,070 and has a Php 7.7
production cost/kg. The result was followed by the carbaryl with a generated ROI of 273.4 %.
48
The least net income was obtained from the MMSU Bio-In 6WP50 and MMSU Bio-In 7WP50
with Php 208,242 and with 226.9% ROI thus it has a Php 9.2 production cost/kg.
Table 29. Cost and return analysis of growing eggplant per hectare sprayed with different
bioinsecticides.(On station)
TREATMENT/
BIOINSECTICIDE
GROSS
INCOME
(Php)
PRODUCTION
COST
(Php)
NET
INCOME (per
hectare)
ROI UNIT OF
PRODUCTION
COST
(Php/kg)
MMSU Bio-In
6WP50 300,000 91,758 208,242 226.9
9.2
MMSU Bio-In
7WP50 300,000 91,758 208,242 226.9
9.2
MMSU Bio In 6 360,000 91,930 268,070 291.6 7.7
MMSU Bio In 7 306,000 91,693 214,307 233.7 9.0
Control 330,000 88,369 241,631 273.4 8.0
Carbaryl 348,000 93,302 254,698 273.0 8.0
Cost and return analysis of growing tomato using biofungicide products (On Station)
In Table 30, biofungicides as a component in integrated disease management for organic
tomato production, obtained better net income from Php 417,784 to Php 577,283 compared of
not using biopesticide with only Php 186,334. Also, higher return on investment was also
realized.
Table 30. Cost analysis per hectare growing tomato on-station using the different formulated
biofungicides products on stations
TREATMENTS YIELD
Kg
GROSS
INCOME
(Php)
NET
INCOME
(Php)
PRODUCTIO
N COST/
HECTARE
(Php)
UNIT COST
OF
PRODUCTION
(Php/kg)
ROI
(%)
MMSU DF2Tm 19,700 492,500 417,784 74,716 15.2 559.2
MMSU DF3Tm 22,200 555,000 480,283 74,716 13.5 642.8
MMSU DF4Tm 26,100 652,500 577,283 74,716 11.5 772.6
MMSU Tricho-
guard
24,100 602,500 528,063 74,437 12.4 709.4
Mancozeb 22,500 562,500 485,434 77,066 17.7 629.9
Control 10,200 255,000 186,334 68,666 26.9 271.4
*Price= Php 25/kg
49
Cost and return analysis of growing garlic using biofungicide products (on station)
Garlic production (on station) using the biofungicides in the production system obtained
higher net income particularly DF4Ga (Php 261,583.00) as MMSU Tricho-guard (Php
261,863.00) lesser unit of production cost per kg of Php 37.7 and the highest PDI of 165.1% and
165.6% respectively.
Table 31. Cost analysis per hectare of growing garlic using the biofungicide product (on-station)
MMSU, 2013-2014.
TREATMENTS YIELD
Kg
GROSS
INCOME
(Php)
NET
INCOME
(Php)
PRODUCTION
COST/
HECTARE
(Php)
UNIT COST
OF
PRODUCTION
(Php/kg)
ROI
(%)
MMSU DF2Ga 4,100 410,000 251,583 158,417 38.6 158.8
MMSU DF3Ga 3,800 380,000 221,583 158,417 41.7 139.9
MMSU DF4Ga 4,200 420,000 261,583 158,417 37.7 165.1
MMSU Tricho-
guard
4,200 420,000 261,863 158,137 37.7 165.6
Mancozeb 3,900 390,000 232,633 157,367 40.4 147.8
Control 3,400 340,000 188,033 151,967 44.7 123.
Comparative economic analysis of organic and farmers practice in tomato production
In the on-farm trials for tomato (Table 32), organic practice obtained higher net income in
Paoay(Php 597,481.10) and Batac ( Php437,491.100). Lower net income was obtained in the
farmers,field of Paoay (Php 487,531.10) and Batac (Php 181,031.10) . Higher ROI was also
realized in organic practice.
Table 32. Comparative cost and return analysis of organic production practice and farmers
practice for organic tomato production in Batac and Paoay
Items Organic practice Conventional practice
Paoay Batac Paoay Batac
I.Return
Price/kg(P) 25.00 25.00 25.00 25.00
Yield(kg) 26,900.00 20,500.00 22,260.00 10,000
Gross income(P) 672,500.00 512,500.00 556,500.00 250,00
II. Variable Cost (P)
A. Labor 41,000.00 41,000.00 39,000.00 39,000.00
B. Materials 32,598.00 32,598.00 28,548.00 28,548.00
C. Fixed Cost 1,420.90 1,420.90 1,420.90 1,420.90
D. Total prod’n. cost 75,018.90 75,018.90 68,968.90 68,968.90
E. Net income/ha 597,481.10 437,491.10 487,531.10 181,031.10
F. ROI 7.96 5.83 7.06 2.62
50
Shelf life of the formulated products
After 6 months of storage no observation in the changes of color, presence of bubbles and
foul odor was noted and in the best biopesticide products against the target pest. In terms of
efficacy preliminary evaluation showed no significant differences when the product was stored
from 1 week to 3 months of storage, although numerically percentage of larva left on the treated
leaf on 1 week of storage was lower by 40% as compared to 1 to 3 months of storage after 16
hours of observation. Further validation is continuous until the product will not deteriorate.
Figure 11 shows the antifungal activity of the botanical formulations was assayed at two
concentrations in the laboratory for their efficacy after 6 months of storage against A. solani
using agar well diffusion method.
Results revealed that botanical product MMSU DF4Tm (20 mm) using 20% was
effective in inhibiting mycelia growth at 6 MAS and were significantly superior over 10% at
11mm only and holds through after 12 months of storage though lower in numerical value. For
both MMSU DF2Tm and MMSU DF3TM, lower values were taken and with decreasing
inhibition with decreased concentration.
Figure 11. In vitro evaluation of botanical formulation against A solani
using 2 concentrations at 6 and 12 months after storage.
After 6 months of storage, MMSU DF3Ga at 20% was most effective in inhibiting A.
porri with24 mm inhibition zone followed by MMSU DF2Ga and MMSU DF4Ga with 19 mm
respectively. Inhibition decreases as the concentration decreases to10%. At 12 MAS, both
concentrations decreased inhibition zones for all the botanical products.
51
Figure 12.In vitro evaluation of botanical formulations using 2 concentrations against A
porri at 6 and 12 months after storage.
52
SUMMARY AND CONCLUSION
Preliminary screening indicated pesticidal effect of the following plants: C. viscosa, A.
mexicana, E.hirta, T. pandacaqui. C. longa, Ocimum sp P.betle A.barbadensis, L.Camara and
garlic waste. Product formulation and reformulation by combining the plants that showed
positive pesticidal effect were done to enhance the toxicity of the different extract. The different
potential products were MMSU Bio-In 3, MMSU Bio-In 6, MMSU Bio-In 7 MMSU Bio-In 8 as
bioinsecticide and MMSU DF4Tm, and MMSU DF4Ga as fungicide in liquid and powdered
form, plus microbial antagonist isolated from extactants. Growth inhibition on the test insect was
noted in regards to reduced number of larval and pupal days as well as the premature mortality
of treated larvae. Reduced disease severity, fruit infection was noted and comparable to
chemical fungicides thus increasing fruit yield over the control. Microbial antagonists identified
in vitro showed a potential antagonistic activity as a potential biocontrol agents against A. solani
in tomato, A. porri and C. duddiae in garlic.
Similarly, insect and disease control was noted when the formulated products were tested
under field conditions both on station and on farm trials. The formulated products had
comparable toxicity with chemical insecticides. Shelf life of the potential products showed that
after six months of storage at room temperature, no sign of deterioration or putrefaction was
observed.
Cost and return analysis showed that the different biopesticide products is comparable
with chemical insecticides and can be combined with other control methods to be more
effective. Goals should focus on the development of pest management products, strategies and
systems which can be an alternative control for conventional synthetic pesticide that is safe for
farmers’ use, and economical.
53
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