3.

0 CHAPTER 3

3.1 MATERIALS AND METHOD

3.1 STUDY SITE

The study was carried out at post office in Ilorin, kwara state, Nigeria.

3.2 PRELIMINARY STUDY

Preliminary study was carried out to determine the presence of Notonectidae In the study site.

Their abundance were determined and the statistics would be taken and the collected

`Notonectidae were transported to the laboratory after it is from different micro aquatic habitats

in the same location. The collected predators would be identified for confirmation in the

laboratory with aid of field guide by Garber and Gabriel (2002). A number ofanopheles (prey)

would be disclosed to the predator prior to the study proper to ascertain Back swimmer as

predator.

3.3 NOTONECTIDAE SAMPLING AND IDENTIFICATION

Notonectidae will be collected from the study site: Asa River, along post office, Ilorin, kwara

state, Nigeria. They will be sampled, using standard dipper of 300ml. collected predators were

put in labeled buckets, filled with water from the site of collection. They will then be transported

to the University of Ilorin research laboratory for morphological identification to the family level

using identification key. Notonectidae were used for the work, after starving some for 24 hours

before the experiment but without starving others.

3.4 ANOPHELES COLLECTION

Anopheles (prey) were collected from the mosquitoes breeding sites, Asa river, along post office,

Ilorin, kwara state, Nigeria. Anopheles larvae were distinguished or identified by their

positioning. They lay parallel to the surface. They were collected using dipper along with water

from the location, they transported to the University of Ilorin research laboratory. The larvae

collected are sorted to different instars, using pipette. For sorting, the sizes of the capsule are

considered. Till the experiment begins, the larvae are given yeast and powdered biscuit.

3.5. 24 HOURS EVALUATION DESIGN FOR THE COMPARISON OF PREDATORY

ABILITY OF NOTONECTIDAE DURING THE DAY AND AT NIGHT.

An experiment will be setup to compare the rate of feeding of Notonectidae on anopheles during

the day and at night. The experiment were carried out in three different ratios (1 : 50, 2: 50 and

5:50) for both starved and unstarved Notonectidae. In each of the ratios, three different replicates

will be for the starved and another three for the unstarved and a control, making a total set up of

19. 1, 2, and 5Notonectidae were offered 50 Anopheles in the ratio of 1: 50, 2: 50 and 5:50

respectively. The number of anopheles consumed will be noted at interval of 3 hours of

predation. The number of preys consumed will be replenished in other to keep the prey density

constant. The PH, dissolved oxygen and electricity in due finity were noted. Predating rate

between 7:00am – 7:00pm for the day and 7:00 pm – 7: 00 am for the night will be monitored

and compared.
3.6 24 HOURS EVALUATION DESIGN FOR COMPARING THE PREDATING RATE

OF STARVED AND UNSTARVED NOTONECTIDAE ON ANOPHELES.

There will be experiment to compare the rate at which starved and unstarved Notonectidae will

prey when exposed in ratios 1:50, 2:50 and 5: 50 for anopheles and Notonectidae respectively,

each ratio having three different replicators for starved and unstarved. Compiled with a control,

there was a total setup of 19. The number of anophelesconsumed by starved and unstarved

Notonectidae will be noted separately and replenish to keep the number of anopheles the same.

PH dissolved oxygen and electrical conductivity were measured. The experiment runs for 24

hours (i.e., 7:00am – 7: 00am the next day). The rate at which starved Notonectidae prey were

compared with that of the unstarved.

3.7 24 HOURS EVALUATION DESIGN FOR COMPARING THE EFFECT OF

DENSITY (NUMBER) OF NETONECTIDAE IN THEIR RATE OF CONSUMPTION AS

PREDATOR OF ANOPHELES.

An experiment were be set to compare the rate at which nectonectidae at their different density

(numbers) were prey on the same (fixed) number of anopheles The experiment will use three

different densities (numbers) of netonectidae to a fixed number of anopheles. The numbers/

density of netonectidae are 1, 2 and 5, each to a fixed number of anopheles which will be 50. 50

anopheles will be exposed to 1, 2 and 5 netonectidae 1.e 1:50, 2: 50, and 5: 50 , with each of the

ratios having 3 replicators in their starved and unstarved fashions. The numbers of anopheles

consumed in each replicates of each ratios were be recorded and replenished to leap the

anopheles at 50 although. The record is taken at the interval of 3 hours. The experiment runs
from 7: am – 7:am the next day, marking its duration to be 24 hours the next day, marking its

duration to 24 hours. The consumption rate of the ratios were be compared.

3.8 STATISTICAL ANALYSIS

A two-way analysis of variance (ANOVA) and Turkey’s multiple comparison test was used to

evaluate the significant differences in the feeding rates and feeding habits of the Notonectidea,

and computer program Windows 10 Excel 365 with least significant difference (LSD) was used

to compare means at p<0.05(statistically significant).

4.0 CHAPTER FOUR

4.1 RESULTS AND ANALYSIS

Figure 1: Shows the graphical illustration of the feeding habits of starved and unstarved

Notonectidea.
Figure 2: shows the feeding rates of starved and unstarved Notonectidea at 3 hours regular

intervals.
 Fig (1-2) above are the graphical illustration of the feeding habits and feeding rate of both

starved and unstarved Notonectidea with respect to mosquitoes at the rates of 1:50, 2:50 and 5:50

respectively. From figure 1 above, it can be deduced that, for 1:50 starved Notonectidea,

between, 10pm to 1am, they had the greatest feeding habits as compared to other feeding

duration i.e they feed most at night. Between 4pm to 7pm, their feeding habits were increased

but lesser as compared to their feeding habit between 10pm to 1 am. Between 7pm to 10pm and

4am to 7 am, their feeding habits were at the same rates. Between 1pm to 4pm, their feeding

habits were lesser as compared to their feeding habits between 7pm to 10pm and 4am to 7 am.

Between 10am to 1 pm and 1am to 4 am, their feeding rates were also equal. They had the least

feeding rates between 7am to 10am i.e. they feed less in the morning. For 1:50 unstarved

Notonectidea, between 1pm to 4 pm,4pm to 7pm and 1am to 4 am, they had the greatest feeding

habits as compared to other feeding durations i.e. they feed more during the day and middle part

of the night. Between 7pm to 10pm, their feeding habits also increased but lesser as compared to

their feeding habits between 1pm to 4 pm, 4pm to 7pm and 1am to 4 am respectively. Between

10pm to 1 am, their feeding habits also increased but lesser as compared to their feeding habits

between but lesser as compared to their feeding habits between7pm to 10pm. between 10am to 1

pm was lesser as compared to their feeding habits between 10pm to 1 am. Between 7am to 10am

and 4am to 7 am, their feeding rates were equal and they had the least feeding rates i.e. they feed

less in the morning.

In the realm of 2:50 starved Notonectidea, between, 7pm to 10pm, they had the greatest

feeding habits as compared to other feeding duration i.e. they feed more at night. Between 7am

to 10am, their feeding habits were increased but lesser as compared to their feeding habit

between7pm to 10pm. between 1pm to 4pm, their feeding habits were lesser as compared to their
feeding habit to between 7am 10am. Between10pm to 1 am and 4am to 7 am, their feeding rates

were equal and lesser as compared to their feeding habit between1pm to 4pm. between10am to 1

pm and 4pm to 7pm, their feeding rates were equal and lesser as compared to their feeding habit

between10pm to 1 am and 4am to 7 am respectively. The least feeding habits were observed

between 1am to 4am i.e. they feed less in the mid night. For 2:50 unstarved Notonectidea,

between, 1pm to 4pm, they had the greatest feeding habits as compared to other feeding

durations i.e. the feed more during the day. Between 7am to 10am, 1am to 4am and 4am to 7am,

their feeding rates were equal and lesser as compared to their feeding habit between 1pm to 4pm.

between 10am to 1pm, their feeding habits were lesser as compared to their feeding habit

between 7am to 10am, 1am to 4am and 4am to 7am respectively. Between 4pm to 7pm, their

feeding habits were lesser as compared to the feeding habit between 10am to 1 pm. between

10pm to 1 am, feeding habits were lesser as compared to the feeding habit between 4pm to 7pm

and they had the least feeding habits i.e they feed less in the evening.

In the realm of 5:50 starved Notonectidea, between 7am to 10am, they had the greatest

feeding habits as compared to other feeding duration i.e. they feed more in the morning. Between

7pm to 10pm, their feeding habits were increased but lesser as compared to their feeding habit

between7am to 10am. Between 4pm to 7pm, their feeding habits were lesser as compared to their

feeding habit between 7pm to 10pm. between 10am to 1 pm their feeding habits were lesser as

compared to their feeding habit between 4pm to 7pm. between 1pm to 4pm, their feeding habits

were lesser as compared to the feeding habit between 10am to 1 pm. Between 4am to 7 am their

feeding habits were lesser as compared to the feeding habit between 1pm to 4pm. Between 1am

to 4 am, their feeding habits were lesser as compared to the feeding habit between 4am to 7 am.

Between 10pm to 1 am, feeding habits were lesser as compared to the feeding habit between 1am
to 4 am and they had the least feeding habits i.e. they feed less in the middle of the night. For

5:50 unstarved Notonectidea similar to 2:50, between 7am to 10am, they had the greatest feeding

habits as compared to other feeding durations i.e. they feed more during the day. Between 4pm

to 7pm, their feeding habits were increased but lesser as compared to their feeding habit between

7am to 10am. Between 10am to 1 pm their feeding habits were lesser as compared to the feeding

habit between 4pm to 7pm. between 1pm to 4pm their feeding habits were lesser as compared to

the feeding habit between 10am to 1 pm. Between 10pm to 1 am, feeding habits were lesser as

compared to the feeding habit between 1pm to 4pm. between 7pm to 10pm feeding habits were

lesser as compared to the feeding habit between 10pm to 1 am. Between 1am to 4 am, their

feeding habits were lesser as compared to the feeding habit between 7pm to 10pm and they had

the least feeding habits i.e they feed less in the middle of the night.

From figure 2, At 3hrs, there was no significant difference in the feeding rates between

Starved 1:50 vs. Unstarved 1:50 and Starved 1:50 vs. Starved 2:50. But there was a reasonable

significant difference in the feeding habits and feeding rates between Starved 1:50 vs. UnStarved

2:50, Starved 1:50 vs. Starved 5:50, Starved 1:50 vs. UnStarved 5:50, Starved 1:50 vs. Control

and UnStarved 1:50 vs. Starved 2:50. But no significant difference was recorded in feeding rates

between UnStarved 1:50 vs. UnStarved 2:50. There was a reasonable significant difference in the

feeding rates between UnStarved 1:50 vs. Starved 5:50 and UnStarved 1:50 vs. UnStarved 5:50

but no significant difference was recorded in the feeding rates between UnStarved 1:50 vs.

Control and Starved 2:50 vs. UnStarved 2:50. A reasonable significant difference was observed

in feeding rates between Starved 2:50 vs. Starved 5:50, Starved 2:50 vs. UnStarved 5:50,

Starved 2:50 vs. Control, UnStarved 2:50 vs. Starved 5:50, UnStarved 2:50 vs. UnStarved 5:50,
UnStarved 2:50 vs. Control, Starved 5:50 vs. UnStarved 5:50, Starved 5:50 vs. Control and

UnStarved 5:50 vs. Control.

At 6 hours, there was no record of significant difference in the feeding rates among

Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 2:50 and Starved 1:50 vs. UnStarved

2:50. There was significant difference in the feeding rates among Starved 1:50 vs. Starved 5:50,

Starved 1:50 vs. UnStarved 5:50 and Starved 1:50 vs. Control group but no significant difference

was recorded in the feeding rates between UnStarved 1:50 vs. Starved 2:50. A reasonable

significant difference was observed in the feeding rates among the following groups of

UnStarved 1:50 vs. UnStarved 2:50, UnStarved 1:50 vs. Starved 5:50, UnStarved 1:50 vs.

UnStarved 5:50 and UnStarved 1:50 vs. Control but no significant difference was recorded in the

and feeding rates between Starved 2:50 vs. UnStarved 2:50. A reasonable significant difference

between was observed in the and feeding rates between Starved 2:50 vs. Starved 5:50, Starved

2:50 vs. UnStarved 5:50, Starved 2:50 vs. Control, UnStarved 2:50 vs. Starved 5:50, UnStarved

2:50 vs. UnStarved 5:50, UnStarved 2:50 vs. Control, Starved 5:50 vs. UnStarved 5:50, Starved

5:50 vs. Control and UnStarved 5:50 vs. Control respectively.

At 9 hours, there was no record of significant difference in the feeding rates among

Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 2:50 and Starved 1:50 vs. UnStarved

2:50 but records of significant difference was observed in the feeding rates among Starved 1:50

vs. Starved 5:50, Starved 1:50 vs. UnStarved 5:50, Starved 1:50 vs. Control, UnStarved 1:50 vs.

Starved 2:50, UnStarved 1:50 vs. UnStarved 2:50, UnStarved 1:50 vs. Starved 5:50, UnStarved
1:50 vs. UnStarved 5:50 and UnStarved 1:50 vs. Control groups respectively but no record of

significant difference was observed in the feeding rates of Starved 2:50 vs. UnStarved 2:50,

Starved 2:50 vs. UnStarved 5:50, UnStarved 2:50 vs. UnStarved 5:50 and Starved 5:50 vs.

UnStarved 5:50. A reasonable significant difference was observed in the feeding rates among the

following groups of UnStarved 1:50 vs. Control, Starved 2:50 vs. Starved 5:50, Starved 2:50 vs.

Control, UnStarved 2:50 vs. Starved 5:50, UnStarved 2:50 vs. Control, Starved 5:50 vs. Control

and UnStarved 5:50 vs. Control respectively.

At 12 hours, there was no record of significant difference in the feeding rates among

Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 2:50 and Starved 1:50 vs. UnStarved

2:50 but a reasonable significant difference was observed in the feeding rates among the

following groups of Starved 1:50 vs. Starved 5:50, Starved 1:50 vs. UnStarved 5:50, Starved

1:50 vs. Control, UnStarved 1:50 vs. Starved 2:50, UnStarved 1:50 vs. UnStarved 2:50,

UnStarved 1:50 vs. Starved 5:50, UnStarved 1:50 vs. UnStarved 5:50 and UnStarved 1:50 vs.

Control respectively. there was no record of significant difference in the feeding rates among

Starved 2:50 vs. UnStarved 2:50, Starved 2:50 vs. UnStarved 5:50, UnStarved 2:50 vs.

UnStarved 5:50 and Starved 5:50 vs. UnStarved 5:50 but a reasonable significant difference was

observed in the feeding rates among the following groups of Starved 2:50 vs. Starved 5:50,

Starved 2:50 vs. Control, UnStarved 2:50 vs. Starved 5:50, UnStarved 2:50 vs. Control, Starved

5:50 vs. Control and the UnStarved 5:50 vs. Control.

 At 15 hours, there was no record of significant difference in the feeding rates among

Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 2:50, Starved 1:50 vs. UnStarved

2:50, UnStarved 1:50 vs. UnStarved 2:50, UnStarved 1:50 vs. UnStarved 5:50, Starved 2:50 vs.

UnStarved 2:50, Starved 2:50 vs. UnStarved 5:50, UnStarved 2:50 vs. UnStarved 5:50 and

Starved 5:50 vs. UnStarved 5:50 but a reasonable significant difference was observed in the

feeding rates among the following groups of Starved 1:50 vs. Starved 5:50, Starved 1:50 vs.

UnStarved 5:50, Starved 1:50 vs. Control, UnStarved 1:50 vs. Starved 2:50, UnStarved 1:50 vs.

Starved 5:50, UnStarved 1:50 vs. Control, Starved 2:50 vs. Starved 5:50, Starved 2:50 vs.

Control, UnStarved 2:50 vs. Starved 5:50, UnStarved 2:50 vs. Control, Starved 5:50 vs. Control

and UnStarved 5:50 vs. Control.

At 18 hours, there was no record of significant difference in the feeding rates among

Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 2:50, Starved 1:50 vs. UnStarved

2:50, Starved 2:50 vs. UnStarved 2:50, Starved 2:50 vs. UnStarved 5:50, UnStarved 2:50 vs.

UnStarved 5:50 and Starved 5:50 vs. UnStarved 5:50 but reasonable significant difference was

observed in the feeding rates among the following groups Starved 1:50 vs. Starved 5:50, Starved

1:50 vs. UnStarved 5:50, Starved 1:50 vs. Control, UnStarved 1:50 vs. Starved 2:50, UnStarved

1:50 vs. UnStarved 2:50, UnStarved 1:50 vs. Starved 5:50, UnStarved 1:50 vs. UnStarved 5:50,

UnStarved 1:50 vs. Control, Starved 2:50 vs. Starved 5:50, Starved 2:50 vs. Control, UnStarved

2:50 vs. Starved 5:50, UnStarved 2:50 vs. Control, Starved 5:50 vs. Control and UnStarved 5:50

vs. Control.
 At 21 hours, there was no record of significant difference in the feeding rates among

Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 2:50, Starved 1:50 vs. UnStarved

2:50, UnStarved 1:50 vs. UnStarved 2:50, Starved 2:50 vs. UnStarved 2:50, Starved 2:50 vs.

UnStarved 5:50 and Starved 5:50 vs. UnStarved 5:50 but reasonable significant difference was

observed in the feeding rates among the following groups Starved 1:50 vs. Starved 5:50, Starved

1:50 vs. UnStarved 5:50, Starved 1:50 vs. Control, UnStarved 1:50 vs. Starved 2:50, UnStarved

1:50 vs. Starved 5:50, UnStarved 1:50 vs. UnStarved 5:50, UnStarved 1:50 vs. Control, Starved

2:50 vs. Starved 5:50, Starved 2:50 vs. Control, UnStarved 2:50 vs. Starved 5:50, UnStarved

2:50 vs. UnStarved 5:50, UnStarved 2:50 vs. Control, Starved 5:50 vs. Control and UnStarved

5:50 vs. Control.

At 24 hours, reasonable significant difference was observed in the feeding rates among

the following groups Starved 1:50 vs. UnStarved 1:50, Starved 1:50 vs. Starved 5:50, Starved

1:50 vs. UnStarved 5:50, Starved 1:50 vs. Control, UnStarved 1:50 vs. Starved 5:50, UnStarved

1:50 vs. UnStarved 5:50, UnStarved 1:50 vs. Control, Starved 2:50 vs. Starved 5:50, Starved

2:50 vs. UnStarved 5:50, Starved 2:50 vs. Control, UnStarved 2:50 vs. Starved 5:50, UnStarved

2:50 vs. UnStarved 5:50, UnStarved 2:50 vs. Control, Starved 5:50 vs. Control, UnStarved 5:50

vs. Control but was no record of significant difference was observed in the feeding rates among

Starved 1:50 vs. Starved 2:50, Starved 1:50 vs. UnStarved 2:50, UnStarved 1:50 vs. Starved

2:50, UnStarved 1:50 vs. UnStarved 2:50, Starved 2:50 vs. UnStarved 2:50 and Starved 5:50 vs.

UnStarved 5:50 respectively. Thus, this implies that, 5:50 starved Notonectidea had the highest

feeding rates while 1: 50 unstarved Notonectidea had the lowest feeding rate throughout the 24

hours experimental duration period.

 CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND REFERENCES

5.1 DISCUSSION

Aquatic mosquito predators have direct role in immature predation and sub lethal effect

through physiological and ovipositor deterrence. Among the aquatic predators are the species of

Notonectidae. Notonectidae is a cosmopolitan family of aquatic insects in the order Hemiptera,

commonly called backswimmers because they swim "upside down" (inverted). They are all

predators. They are similar in appearance to Corixidae (water boatmen), but can be separated by

differences in their dorsal-ventral coloration, front legs, and predatory behavior. (Julianna et al.,

2012). Xenobiotics also play a major role in inducing genetic variations among populations of

mosquitoes which are responsible for inducing different resistance mechanisms such as the

exposure of mosquito larvae to fluoranthene and copper increased the level of resistance to

permethrin the most widely used insecticide against mosquitoes (Poupardin et al., 2008). It has

been observed that the incidence of increasing resistance in different mosquitoes is directly

associated with increased urbanization.

Unfortunately, most of the arthropods pests including mosquitoes have developed

resistance against the commonly used chemical insecticides. There are four main mechanisms

through which these pests have developed resistance. The first one is kdr mediated target site

insensitivity which is also known as knockdown resistance. This type of resistance is specifically

identified in mosquitoes, which is caused by the mutations in Voltage-gated Sodium Channel

(VGSC) genes. So far, there are 11 VGSC mutations identified in the mosquitoes only. The
second mechanism is the enhanced metabolic detoxification which results in the detoxification of

chemical insecticides through various metabolic activities. For instance, the mosquito

populations which became resistant to DDT by adapting the mechanism of enhanced metabolic

detoxification also became resistant to deltamethrin due to cross resistance. The third mechanism

is the development of a thick cuticle layer which reduces the penetration of insecticide in the

body. The results of this experiments has shown that there were variations in the feeding habits

of Notonectidea at different durations ranging from 7am - 10am, 10am – 1pm, 1pm – 4pm, 4pm

– 7pm, 7pm- 10pm, 10pm – 1am, 1am – 4am, 4am – 7am respectively.

We also affirmed that, the 1:50 starved Notonectidea feed most at night and they feed less

in the morning. For 1:50 unstarved Notonectidea, they feed more during the day and middle part

of the night and they feed less in the morning. In the realm of 2:50 starved Notonectidea, they

feed more at night while they feed less in the mid night. For 2:50 unstarved Notonectidea, they

feed more during the day and they feed less in the evening. In the realm of 5:50 starved

Notonectidea, they feed more in the morning and feed less in the middle of the night. For 5:50

unstarved Notonectidea similar to 2:50, they feed more during the day and feed less in the

middle of the night.

In this present study, throughout the 24 hour of the record observations, it was observed

that, there was a sequential increase in the feeding habits of Notonectidea in following trend

from 1:50 unstarved Notonectidea, 1:50 starved Notonectidea, 2:50 unstarved Notonectidea, 2:50

starved Notonectidea, 5:50 unstarved Notonectidea to 5 :50 starved Notonectidea.. Thus, this

implies that, 5:50 starved Notonectidea had the highest feeding rates while 1: 50 unstarved

Notonectidea had the lowest feeding rate throughout the 24 hours experimental duration period.
5.2 CONCLUSION

From the outcome of this present study, we can conclude on the fact that, the more the number of

starved Notonectidea, the better the efficacy of the control of mosquitoes, therefore reducing the

outbreak of malaria while the lesser the number of starved Notonectidea, the lesser the efficacy

to the control of mosquitoes. This also implies that, starved predators are best use in the control

of mosquito than the unstarved predators.