Control study of Musca domestica (Diptera, Muscidae) in Misan Province

Background Houseflies are the most common type of Diptera, specifically Muscidae, worldwide, representing more than 90% of all species. This family has over 170 genera and 4200 species, but a few are of medical significance. This study aimed to estimate and assessing the measures to control and prevent grow-up inside houses and flying of the housefly ( Musca domestica Linnaeus, 1758) in Misan. Methods The study occurred over 12 months, from December 2020 to December 2021. Using plastic containers, Musca domestica were collected from all potential breeding sites in the study region (inside and around houses). Sticky oil paper and traps were used to collect the insects. The collected insects were transferred to sealed plastic containers and then to the laboratory of the Department of Microbiology. Out of 200 randomly selected houses, 150 (75%) contained insects. Light traps and sticky oil papers were the most effective control measures, with 26.7% and 25.9% of the Musca domestica collected from these methods, respectively. The ratio of male (233) to female (456) Musca was 1:2, with a significant difference between the frequencies (P<0.05). A large population of houseflies was collected during the hot season (501, 72.7%), whereas fewer Musca were collected during the cold months (188, 27.3%), with a strongly significant difference (P<0.05). The percentage of HI was 54.4%, the CI was 21.9%, and the BI was 79.9%. The overall larval densities (LD) were at a medium level. Conclusions Misan has a high density of Musca domestica, with females being more prominent than males. Hot climate, humid sites, and dirty places are responsible for the breeding of houseflies. The overall larval density was medium. Therefore, the risk of transmitting infectious diseases by houseflies is high within the boundaries of Misan province, and effective control parameters should include measures like light traps and sticky oil.


Introduction
Musca domestica (Order Diptera, family Muscidae) is a major concern for human health because they are vectors of many tropical and subtropical diseases. 1,2Houseflies are abundant in regions like dirty places; they prefer warm environments and occur in moist regions during the daytime. 3They frequently transit around dirty places, animals, and food sources, defecating during feeding, making them ideal disease vectors for spreading microorganisms.Several studies have reported that these vectors transmit several communicable illnesses via pathogens collecting on their body parts, such as female laying eggs on decomposed materials. 4,5Houseflies can transmit leprosy, anthrax, tuberculosis (TB), dysentery, typhoid, diphtheria, and gastrointestinal parasites.Additionally, they play a role as mechanical vectors or intermediate hosts for nematodes and cestodes. 6][9][10][11][12][13] Mullen and Durden 14 and Sales et al. 15 identified different parasites and worm eggs in fly feces, including Diphyllobothrium sp., Trichuris trichiura, Hymenolepis sp., Strongyloides stercoralis, Ascaris lumbricoides, Enterobius vermicularis, Toxocara cani, Giardia sp., Taenia sp., Trichomonas sp., Entamoeba histolytica, and fungi.
Houseflies can be controlled through physical and chemical means.7][18] Traditionally, control processes have focused on killing insects by using different insecticides.Environmental treatment involves removing breeding sites through microbiological ovicides, chemical larvicides and pupacides in regions where endemic-borne diseases occur. 17Musca domestica breeds can live in various habitats, including freshwater habitats, water in mangrove forests, septic tanks, domestic waste, desert coolers, indoor and outdoor environments with stagnant water conditions as humidity, clear roots of aquatic plants, and damp places. 18It takes seven to 10 days for Musca to complete its life cycle under good conditions as median temperature and moderate humidity and not rainy weather, while it can take up to two months under poor conditions like hotter temperatures, heavy rain and colder weather.In temperate locations, twelve generations may occur during one year, whereas in the tropics and subtropics, it may take more than 20 generations. 19[22][23] This study aimed to estimate and assess the measures used to control the housefly (Musca domestica Linnaeus, 1758) in Misan.

Ethics approval
The approval was granted by the University of Misan, Faculty of Medicine Committee Board (ID No. 103/Oct 2020).
• Ethical committee approved the selection of private houses for the study: Faculty of Medicine, University of Misan • The approval number: 103

REVISED Amendments from Version 1
The article has been revised, many of the vocabulary and concepts have been modified, and many definitions have been added to the abbreviations that were in the research in the conclusion and introduction, as well as in the method of work and results.A map of Maysan Governorate has been added, with the percentage of collection and presence of house flies during the research period written on it.This amendment to my research came as a result of reading the valuable comments that the reviewers sent to us.

Insect collection
Using plastic containers, Musca flies were collected from all potential breeding sites in the study region (inside and around houses).Sticky oil paper (insect glue snares) (FLYING, China, Cat.No. 15-B) and traps containing light (Moth UV light trap, Japan, Cat.No. DSCF199055) were used to collect the insects by placing papers and traps randomly indoor and outdoor the houses and in the collecting habitats.They were then placed in Petri dishes (ATACO, China, Cat. No. 34809) which were filled with distilled water (BDH, UK, Cat.No. 45550W) according to Alsaad and Kawan 24 and kept till the time of examination and identification (Figure 2).
The collected insects were transferred to sealed plastic containers and then to the laboratory of the Department of Microbiology.The houseflies were identified based on their shapes, morphologies, and sizes using single concave microscope slides which are helpful in fixing insect (AmScope, US, Cat.No. 660LOO) and anatomical microscope (Olympus, Japan, Cat.No. 2033789).The adult male housefly has reddish eyes positioned closely to each other, and spongious mouthparts.The male is 5-8 mm in length and has four dark stripes on a dull gray thorax, on the dorsal side and pronounced upward bends in the fourth longitudinal wing vein.The basal portions of the abdomen are yellowish, especially on both sides' alignment.Typically, males show a greater laterally yellowish color than females.Dark longitudinal bands run along the median dorsal region of the anterior portion of abdominal segments.Adult female houseflies' eyes are more widely separated than males'.In female houseflies, the bends in the fourth longitudinal wing veins are distinctly more upward than in males.The female is 3-8 mm in length, and have a lightly golden checkered abdomen, more so than males, according to Yeates et al. 25 and Geden et al. 26

Larval densities
Three insect indices, including the Breteau Index (BI), Container Index (CI), and House Index (HI), were measured.In BI, If the percentage of >20%, this represents a high risk of transmission while If it <20% is considered low risk for BI.In CI, if percentage >50%, this represents a high risk of disease transmission, whereas if it <50%, this reflects a low risk for CI.In HI, if percentage ≥5%, this represents a high risk of transmission while that <5% consider low risk for HI. 27The density of the larval index description is a combination of HI, CI, and BI and is rated on a scale of 1-9, as shown in Table 1, according to the Queensland Government. 28The indices scoring is subdivided into three groups: LD = 1 for low, LD = 2-5 for medium, and LD = 6-9 for high (Table 1).

Statistical analysis
All findings were analyzed using SPSS Statistics version 22 (IBM Corporation, New York, US).Categorical data were presented as frequencies and percentages.The chi-square (χ 2 ) test was used to describe the association between categorical variables, with a P-value of <0.05 considered statistically significant.

Results
The houses selection was made randomly, the author visited every city of Misan province by car as well as each town and village of that city.When reaching the place, the author went through the ethics process from their Institution.Additionally, I used my identity card of College of Medicine to take permission for placed papers and traps in collecting  regions) Out of 200 randomly selected houses, 150 (75%) contained insects, while 50 (25%) did not have any larvae.
Houses were the location of samples collection.The housefly collected in this study were collected from these houses by the homeowners using traps and sticky papers supplied by the researcher.I distributed papers and traps to the household owner for free, and collected insects at every visit.The householder gave me consent to collect insects from papers and traps that I bought and gave them freely.Houses were selected for being nearby large water sources and farms or many trees, or regions of good conditions for grow-up of houseflies like moderate humidity and medium temperatures.The distribution of Musca in their habitats was as follows: indoors (48, 6.9%), outdoors (115, 16.8%), plastic cups (69, 10.0%), traps (184, 26.7%), oil papers (179, 25.9%), and sewage water (94, 13.7%) with a high statistical significance (P = 0.001).Light traps and sticky oil papers were the most effective control measures, with 26.7% and 25.9% of the Musca collected using these methods, respectively (Table 2).
In this study, the ratio of male (233) to female (456) Musca was 1:2, with a significant difference between the frequencies (P < 0.05) (Table 3 and Figure 3).
The study period was divided into two parts of the year: hot months (March, April, May, June, July, August, September and October) and cold months (November, December, January and February) (Table 4).A large population of houseflies was collected during the hot season (501, 72.7%), whereas fewer Musca were collected during the cold months (188, 27.3%), with a strongly significant difference (P < 0.05).The percentage of HI was 54.4%, the CI was 21.9%, and the BI was 79.9% (Table 5).The overall LD was at a medium level.

Discussion
The presence of houseflies in different habitats revealed their ability to survive in a particular environment and female oviposition preference in that habitat.Depending on the tolerance range of Musca spp., changes in the physiochemical and biotic-biology features of the habitats may make other environments favorable or unfavorable to successful breeding. 29According to Zulkarnaini and Dameria 30 and Madewell et al., 31 when the HI, CI, and BI are greater than 50%, it indicates a high risk of illness spreading, which is very high in that population.This requires public health professionals and government to use multiple tools to control vectors such as houseflies.
Improving environmental sanitation is fundamental for achieving long-term control of houseflies.Sanitation is the mainstay for controlling houseflies in and around farms or homes.Environmental control involves cleaning garbage areas to reduce odors and prevent housefly breeding. 32Insecticides, natural biological suppression of houseflies, proper management of poultry manure, flytraps, ultraviolet light traps, space sprays containing synergized pyrethrins, baits (excellent selective adulticides), Z-9-triclosan, larvicides, and Neporex are effective in controlling houseflies. 33,34e breeding of flies is closely correlated with the food source where flies forage and breed.In settlements, residents throw garbage daily, attracting flies and contributing to their breeding.The density of flies is closely related to the source of infectious diseases.According to Prabowo, 35 M. domestica is found in very high densities in landfills, markets, and kitchens due to the large quantities of food processed in these areas.Arroyo 36 reported that they are found in many chicken farms, garbage areas, and in animal and human feces.
In this study, only M. domestica was collected in each region because it is commonly found in almost all places, especially food waste and dirt from human activities. 37Additionally, the lifecycle of M. domestica requires ingesting a large amount of food, making it closely related to humans themselves. 38 this study, houses were randomly selected, and 150 (75%) contained insects, while 50 (25%) had no larvae.This is similar to reports from Adenusi and Adewoga, 39 who mentioned that epidemiological investigations found houseflies were carriers of intestinal parasites in dirty places and garbage.Azrul 40 and Sigit et al. 41 documented that the flying distance of houseflies can reach between 200 to 2 km.However, the flying distance from population-dense areas is not more than 500 m, and they do not fly continuously as they often stop to forage in the garbage.Ginanjar 42 stated that houseflies are found in high numbers in dwellings with human activities.
In this study, the male-to-female ratio (M:F) was 1:2, and a large population of houseflies was collected during the hot season.HI, CI, and BI percentages were 54.4%, 21.9%, and 79.9%, respectively.4][45][46][47][48][49][50] Many authors have stated that M. domestica has the highest population among flies and is widely found in residential environments, food sources, dirty cooling places, and landfills near human activities. 41,51,52Musca's larval life history parameters are affected by the prey quantities, types of foods, and densities of rearing of housefly.

Conclusions
Misan province has a high density of houseflies, with females being more prominent than males.Hot climates, humid sites, and dirty places are responsible for the breeding of houseflies.The overall larval density was at a medium level.Therefore, the risk of transmission infectious diseases by houseflies is high within the boundaries of Misan province, and effective control measures such as light traps and sticky oil should be implemented.

If applicable, is the statistical analysis and its interpretation appropriate? Yes
Are all the source data underlying the results available to ensure full reproducibility?Yes

Are the conclusions drawn adequately supported by the results? Yes
Competing Interests: No competing interests were disclosed.
Reviewer Expertise: Molecular biology and genetics.General biology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Muhammad Kashif Zahoor
Department of Zoology, Government College University Faisalabad (GCUF), Faisalabad, Pakistan The article has been improved.I don't have further comments.

Is the work clearly and accurately presented and does it cite the current literature? Yes
Is the study design appropriate and is the work technically sound?Yes

If applicable, is the statistical analysis and its interpretation appropriate? Yes
Are all the source data underlying the results available to ensure full reproducibility?Yes

Are the conclusions drawn adequately supported by the results? Yes
Competing Interests: No competing interests were disclosed.
Reviewer Expertise: Entomology, Genetics, Molecular Biology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Introduction:
The very first word ... Musca domestica ○ Different control measures (chemical control and environmental etc.) have been mentioned in this section.You should add some of the drawbacks/loopholes of these measures in favor of or to justify your methods applied in the present study.

Results:
First paragraph needs to be rephrased (in passive voice).
The house selectionwas made randomly, ethical procedure was followed as prescribed by the institution.Out of 200 selected houses, 150 (75%) contained flies, while 50 (25%) did not have any larvae.The collection was made using traps and sticky papers.It is to mention that the houses were selected for being nearby large water sources and farms or trees, or regions of good conditions for grow-up of houseflies like moderate humidity and medium temperatures.The distribution of house flies in these habitats was as follows: indoors (48, 6.9%), outdoors (115, 16.8%), plastic cups (69, 10.0%), traps (184, 26.7%), oil papers (179, 25.9%), and sewage water (94, 13.7%) with a high statistical significance (P = 0.001).Light traps and sticky oil papers were the most effective control measures, with 26.7% and 25.9% of the house flies collected using these methods, respectively (Table 2).Similarly for the number of Musca domestica your sample size is 6, so how df = 6 is possible?
Tables 3, 4 -Your sample size is 2 in Table 3 (male and female category) and Table 4 (Hot months and cold month) while the author gave df = 2 in both Tables -how?
The df values throughout the article indicate that the author need to confirm the sample size/data for statistical analysis.Then also confirm if the subsequent analysis and other value i.e.P-values and X 2 be correct.
The correct statistical analysis is a key to conclude the study in a pragmatic way; otherwise the results couldn't be the actual presentation of what the study was aimed.I suggest you to correct the statistical analysis.

Discussion:
This section looks like the introduction section in the start while in the remaining part of the discussion, the results of the present study are poorly discussed with previous studies.Latest and recent references should be cited.Overall, this section needs to be improved.

Figure 1 .
Figure 1.Misan province map showing the rates of Musca domestica.

Reviewer Report 14
December 2023 https://doi.org/10.5256/f1000research.158405.r223167© 2023 Zahoor M. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Table 2 .
Distribution of Musca domestica in habitats.

Table 3 .
Musca distribution according to sex. 46

Table 4 .
Musca domestica larvae distribution in relation to weather.

Table 5 .
Distribution of Musca according to indices.

Table 2 :
There is confusion in statistical analysis/values.Your sample size is 6 or 7 for number of houses, how is the df 4?

the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? No
You should correlate your collected population of Musca domestica with these parameters.And if it's reported in previous studies you can discuss in the discussion part instead of making it the conclusion of your present study.
; the author concluded that hot climate, humid sites and dirty places are responsible for the breeding of houseflies.Did the author evaluate these parameters in the present study?Where is the data representing these parameters?Competing Interests: No competing interests were disclosed.Reviewer Expertise: Entomology, Genetics, Molecular BiologyI confirm that I

have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.
The author should include the hypothesis of the study.It can be indexed after corrections and necessary additions are made.Make sure that all references are included in the text.

the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests:
No competing interests were disclosed.