Methodology

Statement of the Study
This research investigates the masculinization of the betta splendens (betta fish) through
immersion in different concentrations of commercial honey.
1,) How does the sex of the betta fish determined?

2.) How does honey influence the sex hormones?
3) Is there a significant relationship between the gender of the betta fish and the commercial
honey?
4.) Is there a significant difference on the gender of the betta fish before and after immersion
with commercial honey?
5.) Based on the findings, on what concentrations of commercial honey does the betta fish prefer
and thrives the most?
Methodology
2.1 Subject of the study
Betta splendens (betta fish) are popular because of their color and shapes, specifically the male
betta fish. The betta fish became in demand, but because of the low male percentage of male
betta fish some proposed methods to increase the male population through masculinization.
Following the same method of (A W Soumokil et al 2020) which proved to be effective, the
researchers will randomly choose a breed of betta fish larvae of 5 days (D5) as test subjects. For
two days, the larvae were acclimatized in 50 L of fresh water in two glass aquariums (30x40x45
cm) supplied with aerators. This study employed a laboratory experiment with three treatments
and three replications. Betta larvae are immersed in commercial honey at various concentrations,
namely A (5ml/L), B (6ml/L), and C (7ml/L) treatment and control (without commercial honey).
After acclimatization for 2 days, 7 days (D7) betta fish larvae are placed in the treatment
container, which is a plastic jar (vol. 5 L) filled with 3 L treatment media with a larvae density of
30 ind./treatment. Larvae are immersed in commercial honey for 12 hours before being reared
for 60 days. At the start of larval rearing, egg yolks are served as food, followed by artemia and
mosquito larvae, which are fed ad libitum. Siphoning and water exchanges were performed once
a day in the morning before data collection. For all treatments, the water was replenished by
50%.
2.2 Design
In this study, the researchers will conduct a true experimental design. The researcher will use a
simple random sampling because the researchers want to know the reaction of the randomly
selected betta fish larvae on different concentration of commercial honey and how does it affect
its pre-determined sex.
2.3 Environment
The researchers will be conducting the experiment on the biology laboratory at Babag National
High School, because the researchers will be using some of the equipment such as the
microscope to observe the betta fish larvae. The biology lab is also a suitable environment to
conduct the experiment as the researchers wants to conduct the experiment with minimal
interaction with other organisms as to prevent damaging the larvae and to achieve the desired
results of the study.
2.4 Data Gathering
The researchers will observe the growth and behavior of the betta fish all through out the experiment and
record the data. The variables measured in this study were the survival rate and the percentage of
male and female fish. In the study of (Effendi 1979 as cited by A W Soumokil) similar method is
used.
Rate of Survival (SR). Survival is calculated as the percentage of fish that survive at the
conclusion of rearing divided by the total number of fish larvae at the start of rearing in a
container using the formula from [1], as follows:
𝑆𝑅 (%) =Number of fish at the end research 𝑥 100

Number of fish at the beginning research
To calculate the male and female percentage of betta fish, the calculation can be done using the
formula according to [1], as follows:
𝑴𝒂𝒍𝒆 𝒇𝒊𝒔𝒉 (𝑴)(%)=𝐍𝐮𝐦𝐛𝐞𝐫 𝐨𝐟 𝐦𝐚𝐥𝐞 𝐟𝐢𝐬𝐡 𝒙𝟏𝟎𝟎

𝐍𝐮𝐦𝐛𝐞𝐫 𝐨𝐟 𝐬𝐚𝐦𝐩𝐥𝐞𝐬
𝑭𝒆𝒎𝒂𝒍𝒆 𝒇𝒊𝒔𝒉 (𝑭)(%) = 𝐍𝐮𝐦𝐛𝐞𝐫 𝐨𝐟 𝐟𝐞𝐦𝐚𝐥𝐞 𝐟𝐢𝐬𝐡 𝒙𝟏𝟎𝟎

𝐍𝐮𝐦𝐛𝐞𝐫 𝐨𝐟 𝐬𝐚𝐦𝐩𝐥𝐞𝐬
2.5 Treatment of data

The researchers will use bar graph in presenting the collected data of Survival Rate (SR), male
and female percentage to emphasize the difference of the results then explained thoroughly and
descriptively. In the study of (A W Soumokil 2020) similar method is used. Therefore, the
method on presenting the data is fitting the experiment.
Ethics
Before conducting the experiment on live betta fish larvae, a letter will be sent to Bureau of
Fisheries and Aquatic Resources (BFAR) to conduct this study. Also, before conducting the
experiment the researchers will ask the principal of the school to use the biology laboratory when
it is time to conduct the experiments. Lastly the researchers will ask permission to the classroom
advisor, and since the advisor of the researchers is a major in biology and is expert in this
specific field the researchers will ask for guidance in conducting the experiments and handling
the equipment in the biology laboratory.

Methodology

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Statement of the Study

1,) How does the sex of the betta fish determined?

2.1 Subject of the study

2.4 Data Gathering

𝑆𝑅 (%) =Number of fish at the end research 𝑥 100

𝑴𝒂𝒍𝒆 𝒇𝒊𝒔𝒉 (𝑴)(%)=𝐍𝐮𝐦𝐛𝐞𝐫 𝐨𝐟 𝐦𝐚𝐥𝐞 𝐟𝐢𝐬𝐡 𝒙𝟏𝟎𝟎

𝑭𝒆𝒎𝒂𝒍𝒆 𝒇𝒊𝒔𝒉 (𝑭)(%) = 𝐍𝐮𝐦𝐛𝐞𝐫 𝐨𝐟 𝐟𝐞𝐦𝐚𝐥𝐞 𝐟𝐢𝐬𝐡 𝒙𝟏𝟎𝟎

2.5 Treatment of data

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