Research consultancy

CHAPTER THREE

RESEARCH METHODOLOGY

3.1 Scope of the study

The study will not be restricted to any geographical area. However, experimental bean seeds will be obtained from a known farmer in Kumi District, near Kanyum Comprehensive Secondary School, where the experiment will be conducted. The study will focus only on the efficacy of cow dung ash and chilli pepper powder in controlling bean bruchid weevils. The study experiment shall be conducted for three months, from October 2022 to December 2022, and the finding shall be obtained at the end of the experiment.

3.2 Research design

A research design guides a researcher on which research methods to use to accomplish the study objectives. An experimental Randomized Complete Block Design (CRBD) will be used for the study. The randomized controlled experiment will allow for the rigor to determine whether a cause-effect relationship exists between treatments and outcomes. The study will ensure random allocation of treatments to the different blocks. In this research, RCBD is used because of its level of precision and numerous advantages. It allows for the homogeneity of the setup of the blocks. However, variation may likely exist between blocks, which is the basis for evaluation (Rudhra et al., 2022). This type of design is flexible with respect to the different number of treatments and blocks, and it provides more convincing results than that of a complete randomized design (CRD) due to the introduction of blocking, which allows the computation of unbiased error for specific treatment.

3.3.0 Collection of the chill pepper plant and Cow dung

Fresh chilli pepper plant’s plant parts (leaves, fruits, roots, and bark) will be collected from a known farmer’s garden between 28^th to 29^th September 2022. Different polyethylene peppers will be used to collect different chili plant parts. Personal protective equipment will be put on gloves, masks, and aprons. Mature parts used in this study will be harvested and sorted from other plant parts. The harvested parts will then be transported to a clean place in the school laboratory, where they are picked on subsequent days for drying

Fresh cow dung will be collected from the kraal of the indigenous cattle breeds owned by a local farmer in Kumi District near Kanyum Comprehensive Secondary School. The research will put on personal protective equipment like gloves, gum boots, and an apron before entering the cattle kraal. Only the fresh cow dung will be collected to take a uniform drying period, and no contamination will have occurred to destroy its quality. All the collected cow dung will be put in polyethylene paper and transported to the school laboratory, where it will be picked up in the subsequent days for drying.

3.3.1 Collection of bean bruchids

Live mature mixed-sex bean bruchids weevils will be collected from the infested bean store in Kumi District at Kanyum Comprehensive Secondary School from 4th October to 5^th October 2022. The researcher will get a clean container, and upon reaching the store, personal protective equipment like gloves, mask, and apron will be put on, and bruchids will be picked. After collecting the required number for this study, bruchids will be taken to Kanyum Comprehensive Secondary School Agricultural Laboratory. The bruchid’s colony will be maintained in both Laboratories at room temperature of 21°C to 28°C and relative humidity (RH), which will be measured by thermometer and hygrometer.

3.3.2 Collection of bean seeds

Bean seeds of the NABE 17 variety will be used in this study as they are commonly grown in Uganda (Lukas, 2018). The seeds will be bought from a known farmer in Kumi District near Kanyum Comprehensive Secondary School. This farmer will be chosen to have the ability for the researcher to monitor the harvesting, threshing, and pick the seeds before the insecticide is put. All the broken beans will be sorted from whole grain beans and then dried until the moisture content is reduced to normal. Clean seeds will be brought to Gulu University Multifunctional Research Laboratory and Kanyum Comprehensive Secondary School Agricultural Laboratory. They will be subjected to controlled heating in an oven at 40°C for four hours to ensure that existing eggs, larvae, or live weevils are dead. The heated beans will then be cooled at room temperature of 21°C to 28°C for 2 hours.

3.3.4 Inclusion and exclusion criteria

Beans that have already been harvested and no synthetic chemicals dusted on them with no physical damage will be included in the experiment. Beans that ot yet been harvested mature in the garden or harvested, and synthetic chemicals dusted on them or broken or decaying will be excluded from the study.

3.4 Experimental design

Each treatment will have three replications in a Randomized Complete Block Design (Rosulu et al., 2022). A total of three blocks, each having three treatment levels(replicates), will be used to test for the efficacy of treatments for bean bruchid weevils.

The Randomized Complete Block Design.

Twenty adults live mixed sex bean bruchids will be counted and introduced to each container containing 10g of beans. Each sample will be subjected to a different treatment. Thirty-three containers will be used in the Randomized Complete Block Design experiment. The treatments levels (2g, 4g and 6g) will consist of weighed chilli pepper powder, cow dung ash, and a combination of chilli pepper powder and cow dung ash, and will be added to the beans and shaken until the seeds are fully coated. The control will consist of bean seeds and 10 introduced bruchids, but no chili pepper powder or cow dung ash will be added. The container that will allow experimental material will be put in transparent jars. The tips of jars will be perforated to allow airflow and to confine the bean bruchids in the jars.

3.5.0 Preparation of chili pepper and cow dung

The chili pepper parts and cow dung will be dried using clean materials under sunshine until the moisture content is reduced. The dried materials of chili pepper parts will be pounded using a mortar and pestle into powder. The powder will be stored in airtight plastic containers and put in a refrigerator at four °C to maintain the quality before application.

A clean iron sheet will be used where cow dung will be burnt to ash and left to cool. Cooled cow dung ash will be obtained and packed in a clean container to keep its quality before use as treatment.

3.5.1 Experimental containers and cods

Requ    ired transparent containers with lids will be bought from plastic shop. The containers will be cleaned with soap and water dried in Gulu University Biology laboratory. Dried containers will be perforated by patching holes in the lids and sides  and labelled

Cods will be generated that represents all treatments and blocks using masking tap and a marker like Root powder at treatment level one in block A (RP.1a), Bark powder at treatment level one in block A (Bp. 1a), leaf powder at treatment level one in block A (LP. 1a), combination of bark powder and cow dung ash (BP+CD. 1a) each of these will have replication at three levels of treatments in three blocks. 10g of bean sample will be weighed using digital balance and place in containers.

Three different levels of powders will be weighed and introduced separately into labelled containers.

For treatment level one each powder or powder combination will be introduced like 2g of single powder will be weighed. 1g each double powder combination will be weighed. For four powder combination 0.5g each will be weighed.

10 live mixed sex will be starved for one day in order to start feeding when introduced to the samples  and introduced to the container with weighed treatments and beans

3.5.2 Efficacy test

The set experiment will be monitored and checked on daily bases. The efficacy of the different treatments will be determined based on mortality rate, survival rate, emerged rate, eggs laid, and the number of holes made by bruchids per bean seed for 12 weeks (3 months).

3.6.0 Data quality control

Data quality control will be achieved by increasing control experiment blocking to reduce bias between blocks ensuring proper counting of bean bruchid weevil as either live, dead, young, or emerged, and counting number of holes made by weevils on bean seeds and egg laid. This will be observed by the researcher starting from the experiment setup, and the data obtained will be saved on Google drive with known pass ward

 3.6.1 Statistical analysis

Data obtained will be entered into a data collection sheet and transferred to Microsoft excel. One-way ANOVA statistical method will be used to test the efficacy of individual chili pepper parts on the sample within blocks, providing answers to Objective 1. The same analysis will be done on cow-dang ash to test its efficacy on the sample, thus providing answers to objective three since the two factors (powders in combination) will be applied to the samples to achieve objectives 2 and 4. If a considerable variation is released between blocks, a two-way ANOVA method will be used to test their efficacy on the bruchid population dynamics. Comparisons will be conducted using a least significant difference (LSD) test (p≤ 0.5).

3.7 Ethical consideration

The Gulu University Research Ethical Committee (GUREC) will seek ethical clearance to conduct the study. Permission letter shall be obtained from Gulu University Multifunctional Research Laboratory and Kanyum Comprehensive Secondary School laboratory to experiment.

The laboratory technicians and researchers will be protected by wearing Personal protective equipment like gloves, overalls, glasses, aprons, masks, gum boots, or closed shoes will be used to prevent the effect of chili pepper on their health. All used materials after every work will be washed and stored under safe custody to prevent the chilli pepper effect.

3.8 Limitations/Anticipated problems

The major anticipated problem will be block variation due to temperature and humidity. However, this will be solved by control and using one-way ANOVA in analysis, but where the variation will not be a result of blocking, a two-way ANOVA will be used

They may also be delays in conducting the laboratory experiment. The laboratory may not have all the necessary tools like containers, jars, thermometers, and hygrometers. However, this will be solved by talking to the laboratory assistant in time so that missing tools and equipment can be bought using the proposed budget.

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APPENDIX

Table I: Proposed Budget for research as of September 2022

Table II: Proposed work plan for research