==== Front PLoS One PLoS One plos plosone PLoS ONE 1932-6203 Public Library of Science San Francisco, CA USA 10.1371/journal.pone.0242315 PONE-D-20-30451 Research Article Biology and Life Sciences Zoology Entomology Insects Biology and Life Sciences Organisms Eukaryota Animals Invertebrates Arthropoda Insects Biology and Life Sciences Zoology Animals Invertebrates Arthropoda Insects Physical Sciences Physics Electromagnetic Radiation Light Artificial Light Earth Sciences Geography Human Geography Urban Geography Urban Areas Social Sciences Human Geography Urban Geography Urban Areas Earth Sciences Geography Geographic Areas Urban Areas Earth Sciences Geography Geographic Areas Rural Areas Biology and Life Sciences Zoology Entomology Insects Moths and Butterflies Biology and Life Sciences Organisms Eukaryota Animals Invertebrates Arthropoda Insects Moths and Butterflies Biology and Life Sciences Zoology Animals Invertebrates Arthropoda Insects Moths and Butterflies Biology and Life Sciences Ecology Biodiversity Ecology and Environmental Sciences Ecology Biodiversity Biology and Life Sciences Zoology Entomology Insects Diptera Biology and Life Sciences Organisms Eukaryota Animals Invertebrates Arthropoda Insects Diptera Biology and Life Sciences Zoology Animals Invertebrates Arthropoda Insects Diptera Biology and Life Sciences Zoology Entomology Insects Hemiptera Biology and Life Sciences Organisms Eukaryota Animals Invertebrates Arthropoda Insects Hemiptera Biology and Life Sciences Zoology Animals Invertebrates Arthropoda Insects Hemiptera Impact of artificial light intensity on nocturnal insect diversity in urban and rural areas of the Asir province, Saudi Arabia Effect of artificial light intensity on insect faunaHakami Abdulrahim Refdan ConceptualizationResourcesSoftwareWriting – review & editing1 https://orcid.org/0000-0002-4734-6338Khan Khalid Ali ConceptualizationData curationFormal analysisInvestigationMethodologySupervisionWriting – original draftWriting – review & editing234* Ghramh Hamed A. Formal analysisFunding acquisitionProject administrationResourcesSupervision234 Ahmad Zubair Data curationFormal analysisWriting – original draft235 AL-zayd Adil Ali Ahmad Data curationInvestigationMethodology4 1 Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, Abha, Saudi Arabia 2 Research Center for Advanced Materials Science (RCAMS), King Khalid University, Abha, Saudi Arabia 3 Unit of Bee Research and Honey Production, Faculty of Science, King Khalid University, Abha, Saudi Arabia 4 Biology Department, Faculty of Science, King Khalid University, Abha, Saudi Arabia 5 Biology Department, Faculty of Sciences and Arts, King Khalid University, Dhahran Al-Janoub, Saudi Arabia Farooq Shahid Editor Harran University, TURKEY Competing Interests: The authors have declared that no competing interests exist. * E-mail: khalidtalpur@hotmail.com 1 12 2020 2020 15 12 e024231527 9 2020 30 10 2020 © 2020 Hakami et al2020Hakami et alThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Continuous urban developments have resulted in increased demand for street furniture, one of which is street light columns. Artificial light at night (ALAN) pose significant impacts on insect diversity in urban and rural areas. The ALAN is a significant driver of decline in insect diversity. This study evaluated the impact of light intensity and sky quality at night on insect diversity in rural and urban areas of the Asir province, Saudi Arabia. Insect traps were installed in both areas during night. Light intensity of nearby road lamps was measured using light meter, while sky quality was measured using sky quality meter. Rural areas exhibited low light intensity (10.33 flux/f.candle) and good sky quality (18.80 magnitude/arcsec2). Urban areas exhibited intense light (89.33 flux/f.candle) and poor sky quality (15.49 magnitude/arcsec2). Higher insect diversity was recorded for rural areas where insects belonging to seven orders (i.e., Diptera, Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, Neuroptera, and Dermaptera) were collected. However, insects of four orders (i.e., Diptera, Lepidoptera, Hemiptera, and Neuroptera) were found in urban areas indicating low diversity. Lepidopteran insects were frequently recorded from rural areas indicating they are attracted to artificial light. It is concluded that excessive ALAN and poor sky quality at night disrupt insect biodiversity. Therefore, ALAN and sky quality must be considered responsible for decline in insect biodiversity along with other known factors. Research Center for Advanced Materials Science, King Khalid UniversityRCAMS/KKU/001/20This study was supported by the Research Center for Advanced Materials Science (RCAMS), King Khalid University, Abha Saudi Arabia (RCAMS/KKU/04/20). Data AvailabilityAll relevant data are within the manuscript.Data Availability All relevant data are within the manuscript. ==== Body Introduction Global insect diversity is declining and >40% of the world's insect species could go extinct over the next several decades if declining trends are not halted/reversed [1]. The most common drivers of these declines are intensive agriculture, excessive pesticide use, urbanization, climate change and habitat destruction [1, 2]. Dramatic loss of insect diversity in many parts of the world has attracted the attention of scientists to consider detrimental impacts on human health and livelihoods. Drastic decline of flying insects “Ecological Armageddon” was reported by Hallmann et al. [3]. Malaise traps were used by Hallmann et al. [3] to collect data from an agricultural landscape and >75% decline in insect biomass was recorded over 27 years. This study on decline in insect biodiversity gained significant attention in the scientific community. The substantial loss of insect populations can not only be explained by changes in habitat, climate or land use [3], but there are some other large-scale factors which could also be involved. According to Grubisic et al. [2], artificial light could be an overlooked driver of insect declines. Urbanization has resulted with a new phenomenon associated with cities and towns called ‘light pollution” [4], which has significant impact on animals and insects throughout the world [5]. Increasing artificial light at night (ALAN) is the primary cause of this phenomenon. It is mainly connected to general population growth, industrial development and rising economic prosperity. Installation of lamps with higher luminous efficiency has also played a negative role in insect diversity declines. The ALAN is known to have strong impacts on population dynamics of insects. Globally it has been increasing at an annual rate of 2%–6% over the last decades [6, 7], imposing an unprecedented alteration of natural light regimes and threatening biodiversity [8]. Despite its ubiquity, the importance of artificial light as an agent of global change is often overlooked when analyzing insect population declines. Insects perform many functions and provide essential support in agroecosystems. They provide ecosystem regulation services such as decomposition of organic material, nutrient and energy flow regulation, seed and pathogen dispersal, pollination, control of pests and conservation of biodiversity [9]. Therefore, decreasing number of insects may substantially affect maintenance of these functions and services, with consequences for food production and biodiversity health. Light is an important visual and non-visual cue for insects [10]. More than 60% of all invertebrates are nocturnal [8] and utilize nocturnal light for orientation, navigation, avoidance of predators, location of food and reproduction [11]. Many nocturnal and crepuscular insects use celestial light sources such as stars and the moon as visual cues for dispersal across landscapes [12]. The effect of sky quality and light intensity on nocturnal insects have never been studied in Saudi Arabia. The aim of this study was to examine the impact of ALAN on insect diversity in rural and urban areas. We emphasized the impact of artificial light on insects and compared the effect of sky quality at night and light intensity on nocturnal insects both in urban and rural areas of Saudi Arabia. We investigated the impact of ALAN on insect population declines in light-polluted areas and rural agricultural areas. Materials and methods The experiment was set up according to Manfrin et al. [13] with some modifications. Data relating to diversity of nocturnal flying insect in rural and urban sites were collected to evaluate the impact of light intensity and sky quality on insect diversity in rural and urban areas. Study area This field experiment was conducted in Asir province, Saudi Arabia. No specific permits were required to execute this study and the work did not involve any endangered species. Two different locations, i.e., Mohayil and Abha were selected for this study. We delimited locations according agricultural (Mohayil) and urban area (Abha). Mohayil is characterized by an agricultural landscape, whereas Abha is an urban area. An agricultural farm in Mohayil (18° 12ʹ 24.76ʺ N; 42° 32ʹ 17.51ʺ E) with no prior exposure to artificial light and another location in Abha city (18° 31ʹ 45.31ʺ N; 42° 2ʹ 42.95ʺ E) with light exposure were selected. Environmental conditions Data were collected during February and March, 2020. Light intensity was measured in lux through a light meter, Digitech, QM1587 (Reduction Revolution Pty Ltd, Parramatta, Australia) (Fig 1A). The light meter reads ambient light or direct light from an artificial source and calculates the correct shutter speed and aperture values required to capture an accurate exposure. The sky quality was measured in visual magnitudes per square arc second (magnitude/arcsec2) using sky quality meter (Unihedron, Ontario, Canada) (Fig 1B). Readings of the sky quality meter are inversely proportional to light; hence, higher digits indicate less light intensity. We compared sky quality measurements at different sites quantitatively. 10.1371/journal.pone.0242315.g001Fig 1 Devices used in the experiment. (A) light meter to measure the light intensity from nearby street lights, and (B) sky quality meter to determine the darkness of sky at night. Insect collection and identification Insects were collected from both sites using DynaTrap insect trap (Dynamic Solutions/MII Equipment Inc. Milwaukee, USA). It collects nocturnal flying insects including mosquitoes. Its ultraviolet light and whisper-quiet fan attracts and captures insects without zapping or buzzing. The trap does not use pesticides or chemicals or need an attractant or propane (Fig 2A). Traps were installed from evening to morning (19:00 to 7:00 local time) on every Thursday to Saturday. Sampling was carried out weekly during 1st February to 31st March 2020. The collected specimens were stored in papilon paper in the multipurpose plastic boxes and brought to laboratory for pinning and identification. Order level identification was primarily based on the description by Johnson and Triplehorn [14]. The collected specimens were kept in the Zoological Museum of the College for future reference (Fig 2B and 2C). 10.1371/journal.pone.0242315.g002Fig 2 Experimental steps. (A) DynaTrap insect trap for insect collection, (B) insect trap installed at a rural site and (C) counting and identification of captured insects. Statistical analysis Data were analyzed using the ‘pacman’ package Rinker et al. [15] in R (version 3.6.3; R Core Team, 2019). The analysis included boxplot structure, scatter plot and summary. Only night samples were analyzed for the comparisons of artificial light intensity and sky quality. Results Environmental conditions and measurements Temperature, relative humidity, sky quality and light intensity were 26 to 27°C, 62% to 63%, 18.09 to 19.20 magnitudes/arcsec2 and 10 to 11 flux/f.candle, respectively in rural area. Similarly, a temperature between 18 and 19°C, 70% to 74% relative humidity, 15.18 to 16.10 magnitudes/arcsec2, and 88 to 90 flux/f.candle were recorded for urban area. The altitude of rural area was 530 m above sea level, while urban area is located at an elevation of 2215 m. Table 1 summarizes environmental data of both experimental sites. A significant difference (p ≤ 0.05) was observed in all environmental variables, i.e., light intensity, sky quality, altitude, relative humidity and temperatures between rural and urban area. 10.1371/journal.pone.0242315.t001Table 1 Environmental data of rural and urban sites included in the study. Location Light Intensity (flux/f.candle) Sky quality (msgs/arcsec2) Altitude (m) Relative Humidity (%) Temperature (°C) Mohayil (rural) 10.33 ± 0.58B 18.80 ± 0.61B 530 ± 0.0B 62.33 ± 0.58B 26.67 ± 0.58B Abha (urban) 89.33 ± 1.15A 15.49 ± 0.53A 2215 ± 0.0A 72.33 ± 2.08A 18.67 ± 0.58A Means followed by same letters within a column are statistically non-significant. Light intensity at night was high (89.33 flux/f.candle) with low sky quality (15.49 msgs/arcsec2) in urban area. On the other side, rural area recorded limited manmade intervention (10.33 flux/f.candle) and better sky quality (18.80 msgs/arcsec2) (Fig 3). 10.1371/journal.pone.0242315.g003Fig 3 Scatter plot showing the relationship between light intensity and sky quality at rural and urban areas. Diversity of nocturnal flying insects The insect traps installed at both areas collected insects belonging to seven different orders. Insects from rural site belonged to Diptera, Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, Neuroptera, and Dermaptera orders. The insects collected from urban area belonged to four orders namely Diptera, Lepidoptera, Hemiptera and Neuroptera. The number of insects collected from Diptera, Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, Neuroptera, and Dermaptera orders ranged between 0–43, 37–156, 0–55, 0–5, 11–66, 0–2 and 0–7, respectively in rural area. Similarly, 1–9, 2–9, 0–1 and 0–2 insects were collected for Diptera, Lepidoptera, Hemiptera and Neuroptera, respectively from urban areas. The average number of insects collected from rural area in orders Diptera, Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, Neuroptera and Dermaptera were 18.67, 89.67, 22.00, 1.67, 30.67, 1.00 and 3.00, respectively. In urban area average number of insects belonging to Diptera, Lepidoptera, Hemiptera and Neuroptera were 5.33, 5.00, 0.33 and 0.67, respectively (Fig 4). Lower population of Lepidopterans in urban area suggests that these insects were attracted to the uncountable number of the city white light LEDs. The peak of Lepidopteran insects’ attraction is attained when blue or white light emits shorter wavelengths around 400 nm [16]. It has been observed in Abha that a huge number of insects were attracted to newly-lit areas that have not been lit before. This attracts bats to feed on them. No attraction was observed to the high pressure sodium lamps within the city that are still installed in many roads, although white LEDs are now increasing in an alarming rate. 10.1371/journal.pone.0242315.g004Fig 4 Box plot showing the population of insect orders in response to light intensity and sky quality. (A) box plot for insect diversity in rural area and (B) box plot for insect diversity in urban area. Discussion Nocturnal insects are highly sensitive to light and their behavioral responses like dispersal, foraging, defense against predation, location of food, hosts, oviposition sites, resting sites and mate searching are governed by vision [17–25]. These behavioral responses of insects are very much related with light intensity (adaptation), circadian periodicity and photoperiodism [20, 24]. Nocturnal behavior of insects, their growth and physiology can be disrupted by the use of artificial light at night (ALAN) [23, 24]. However, the reason for this phototactic behavior, especially for ALAN in nocturnal insects remains unclear and may be driven by multiple factors [18, 20, 26]. The effects of environmental (e.g. light intensity, exposure time of light, polarization, weather and season) and physiological (e.g. sex, mating status, age and adaptation to the dark) factors as well as other anthropogenic activities (e.g. intensive agricultural practices), which leads are prominent examples that may affect insect’s population in rural and urban areas [27, 28]. As a consequence, long-term exposure of nocturnal insect communities to ALAN can lead to changes compared to communities that are less exposed to light [29, 30]. Most of the studies on the effect of light intensities have focused on birds, larger vertebrate animals and only a few studies have devoted on insect population affection light intensities. This is first study in the kingdom of Saudi Arabia investigating the impact of ALAN on nocturnal flying insects in urban and rural areas. The current study confirmed our hypothesis that sky quality and artificial light intensity influences composition and abundance of the insects. Phototactic behavior of insects is also affected by sky quality, light quality and external shape of light source [17, 18, 21, 31]. Species diversity and abundance of insects caught by light traps can change according to the light source, for example, LED bulb, compact fluorescent bulb or incandescent bulb [31]. In rural area sky was clear and light intensity from artificial sources was lower than urban areas. This phenomenon proved good for insect diversity and populations [17, 18, 21]. Indeed, natural areas with low-level of artificial light are valuable to maintain healthy ecosystem and balanced insect diversity. In rural area seven insect orders were observed compared to four insect orders in the urban area. Artificial illumination attracted large number of nocturnal flying insects. Light sources function as an ecological trap [32] for many insects. This occurs during swarming events, when very large numbers of population are attracted to artificial light sources [33]. If not killed immediately, insects are often unable to disperse and migrate elsewhere [32, 34]. Several studies have shown how artificial illumination disrupts dispersal patterns in arthropods, confounding natural sources of orientation (e.g., moonlight) and attracting positively phototactic insects [35, 36]. In this study, majority of insects collected in the light traps were of nocturnal nature suggesting that these might be sensitive; thus, more vulnerable to light intensity [37]. In urban areas, increased light intensity at night distracts significant number of insects. This could be a possible reason for the low diversity and the reduced number of insets. These results are in accordance with Horváth et al. [35], Meyer and Sullivan [36]. Moreover, the presence of artificial light may affect the distance that organisms and nocturnal creatures move away [34, 38]. The ALAN has substantial effects on insect fauna, including moths. Macgregor et al. [39] reported that moth abundance was reduced to half and flight activity was 70% greater at lit sites as compared to dark areas. They further reported a considerable reduction in pollen transport by moths at areas with artificial light. Their findings supported the disruptive impact of ALAN on moth activity, which is a one proposed mechanism driving moth declines, and suggest that street lighting may potentially affect pollination of nocturnal invertebrates. In our study, the lepidopteron insects were significantly higher in an agricultural farmland (that has not experienced lighting projects before) than urban area. These findings are in agreement with Macgregor et al. [39] who reported that street lights have disruptive impact on moth behavioral activities. Considering the overall scenario, our results demonstrate that the intensity of light in urban environment might be a potential threat to the nocturnal insects’ diversity. Conclusions Natural darkness at night is beneficial for insect species and entire wildlife. This may be understood by the high diversity of insects in rural areas. On the other hand, well-lit areas at night would negatively affect insect population, and this has been proved in this study. The right type of light and the intensity required for urban life are suggested to be considered carefully. Warm light that emits the wavelengths of yellow light may reduce some of the negative impacts and can reduce the number of insects attracted to irresistible artificial light. Authors would like to acknowledge the support of the Research Center for Advanced Materials Science (RCAMS), King Khalid University, Abha Saudi Arabia for funding this work through research program RCAMS/KKU/04/20. Insect images in Fig 4 were created with BioRender.com. 10.1371/journal.pone.0242315.r001 Decision Letter 0 Farooq Shahid Academic Editor © 2020 Shahid Farooq2020Shahid FarooqThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Submission Version0 13 Oct 2020 PONE-D-20-30451 Impact of artificial light intensity on nocturnal insects found in urban and rural areas of the Asir province, Saudi Arabia PLOS ONE Dear Dr. Khan, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. 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The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ********** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ********** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ********** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ********** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: General comment: The paper reports interesting and important findings on Impact of artificial light intensity on insect population. It is well executed paper, clearly written and sufficiently illustrated. The manuscript contains several minor inconsistencies which are commented in the following lists Introduction: Introduction is a good review of the knowledge on the subject. However, the presentation of the light impact behavior with different factors involved like declining insect population, impact of artificial light, local impact etc., need to be improved/ arranged paragraph wise. Also, the introductory part should be more elaborative with latest references in order to provide more comprehensive presentation of the problem. Recent reports of insect population declines might be representing a global phenomenon in which the many stressors of the Anthropocene are pushing insects over the edge of population viability. This interesting aspect of artificial light use is defiantly stressing insects, we need to understand such patterns on insect decline and mechanisms. the authors must provide for these assertions/assumptions while writing introduction. Methodology: The methodology is succinct presented but correct. The figures are satisfactory, except Figure 1 which is poorly structured. The main purpose of these figures is to show the localization of rural and urban region of study area. Author must replace this with better one. I guess the work is very limited to the number of locations and even why this location is specifically chosen is not well expressed. Also the number of samples replicates have not been mentioned. The devices used to perform light intensity data and insect collection traps are standard one, I did not find the methods of insect identification used in this paper. Results: It is the best part of the manuscript. The results are focused and well presented on the insect population dynamics from both rural and urban areas. Discussion: The discussion consists in a good comparison of the insect populations from both urban and rural areas. However, there is a fewer reference in the discussion section. The authors must provide more recent citations to justify the current study with previous one. Reviewer #2: In the present manuscript the author did an effort to explore the “Impact of artificial light intensity on nocturnal insects found in urban and rural areas of the Asir province, Saudi Arabia”. The subject is really an interesting to read because the growing urbanization is adversely damaging the ecosystem of entire life in the world including insects. Insects are very beneficial for the life security as their contribution in crop pollination, honey production, and predator’s pf several other insect species are very important for the agriculture sector and food availability. In the present study author, collected several insect species form the rural and urban areas in the Asir province of Saudi Arabia by using the latest techniques of insect collection. Present work highlights the importance of urban ecology study with reference to insect population and biodiversity in general. Reviewer #3: The article is well written according to the title. However, the article has some grammatical issues. Therefore, it is my recommendation to revise the manuscript well. I have highlighted some issues here; Abstract Line 19: Change“street light columns” by “street-light columns” Line 23: Change“Light intensity” by “The light intensity” Line 24: Change“light meter” by “a light meter” Line 25: Change“darkness the sky” by “dark the sky” Introduction Line 36: Change“Decline in insect” by “A decline in insect” Line 49: Change“new phenomenal” by “new phenomenon” Line 60: Change“decreasing number” by “the decreasing number” Line 60: Change“maintenance” by “the maintenance” Line 67: Change“Arabia the effect” by “Arabia, the effect” Line 68: Change“ALAN on insect” by “ALAN on the insect” Materials and methods Line 81: Change“An agricultural farm land” by “Agricultural farmland” Line 104: Change“attracts and captures” by “attract and capture” Line 116: Change“boxplot” by “the boxplot” Results Line 157: Change“high pressure sodium” by “high-pressure sodium” Line 157: Change“installed in many” by “installed on many” Line 158: Change“increasing in an alarming” by “increasing at an alarming” Discussion Line 173: Change“maintain healthy” by “maintain a healthy” Line 173: Change“balanced diversity” by “a balanced diversity” Line 177: Change“of population” by “of the population” Line 184: Change“the possible reason” by “a possible reason” Line 188: Change“fauna including” by “fauna, including” Line 194: Change“in an agricultural farm land” by “in agricultural farmland” Line 196: Change“have disruptive” by “have a disruptive Conclusions Line 200: Change“affect insect” by “affect the insect” Line 202: Change“ Warm light” by “The warm light” On the other hand, there is repetition of figure 5 and table 2, therefore use one thing. Moreover, following reference could be cited; Haider, I., Akhtar, M., Noman, A. and Qasim, M., 2020. Population Trends of Some Insect Pests of Rice Crop on Light Trap and its Relation to Abiotic Factors in Punjab Pakistan. Pakistan Journal of Zoology ********** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No Reviewer #3: Yes: Muhammad Qasim Istitute of Insect Sciences Zhejiang University [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. Attachment Submitted filename: MS_LIGHT_INSECT_27_09_2020.docx Click here for additional data file. 10.1371/journal.pone.0242315.r002 Author response to Decision Letter 0 Submission Version1 27 Oct 2020 Dear Dr. Shahid Farooq. Academic Editor PLOS ONE Subject: SUBMISSION OF REVISED MANUSCRIPT (PONE-D-20-30451) Thank you very much for your comments regarding our manuscript titled: “Impact of artificial light intensity on nocturnal insects found in urban and rural areas of the Asir province, Saudi Arabia”. I have gone through all the changes suggested by the academic editor and reviewers. They pointed out very important points and provided very useful suggestions. I gave revised the manuscript accordingly. I have accepted all the changes suggested by the reviewers. Comments and responses of all the critique are presented in revised manuscript. The detail of responses to the editor/ reviewer’s comments is provided below in a tabular form. ACADEMIC EDITOR 1 The site selection needs clarity. One which basis these sites were selected? The delimitation of the site selection is provided in the text 2 Figure 1 must be replaced with a better one. Replaced 3 Introduction must be rearranged as suggested by the reviewers. Rearranged accordingly. 4 There should be more reference citations in the discussion section to support your claims. Done. Latest reference included 5 In acknowledgment figure 2 is about sky quality meter instead of insects. It is corrected and now this figure 2 replaced with figure 5. 6 Can the scale of Figure 5B be changed? Scale of Fig.5B Changed from 50 to 15. Journal requirements 1 Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. Checked 2 We suggest you thoroughly copyedit your manuscript for language usage, spelling, and grammar. If you do not know anyone who can help you do this, you may wish to consider employing a professional scientific editing service The language of the manuscript has been edited by Dr. Philip Smith, Uludağ University, Turkey. The language editing certificate is attached as other file. 3 In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why? No specific permits were required to execute the study and the work did not involve any endangered species 4 We note that Figure 1 in your submission contains map images which may be copyrighted. The map was created by R statistical environment with open source shape files. Therefore, the figure is not copyrighted. The detailed explanation is provided in the legend of Figure 1. REVIEWER-1 1 Introduction: T Introduction is a good review of the knowledge on the subject. However , the presentation of the light impact behavior with different factors involved like declining insect population, impact of artificial light, local impact etc., need to be improved/ arranged paragraph wise. Also, the introductory part should be more elaborative with latest references in order to provide more comprehensive presentation of the problem. Recent reports of insect population declines might be representing a global phenomenon in which the many stressors of the Anthropocene are pushing insects over the edge of population viability. This interesting aspect of artificial light use is defiantly stressing insects, we need to understand such patterns on insect decline and mechanisms. the authors must provide for these assertions/assumptions while writing introduction. The introduction is thoroughly rearranging with the additions of latest reference as per the suggestion of referee. 2 Methodology: The methodology is succinct presented but correct. The figures are satisfactory, except Figure 1 which is poorly structured. The main purpose of these figures is to show the localization of rural and urban region of study area. Author must replace this with better one. I guess the work is very limited to the number of locations and even why this location is specifically chosen is not well expressed. Also the number of samples replicates have not been mentioned. The devices used to perform light intensity data and insect collection traps are standard one, I did not find the methods of insect identification used in this paper . Methodology is sufficient and the query of reviewer regarding the site delimitation is added in the text. The number of samples replicates have already mentioned in text Insect identification part is corrected in MS with appropriate references. 3 Results: It is the best part of the manuscript. The results are focused and well presented on the insect population dynamics from both rural and urban areas. The section is further improved as suggested by reviewer-1 4 Discussion: The discussion consists in a good comparison of the insect populations from both urban and rural areas. However, there is a fewer reference in the discussion section. The authors must provide more recent citations to justify the current study with previous one This section also rearranged as per the advice of reviewer and also the latest references have been provided in the justification of the present study REVIEWER-2 1 In the present manuscript the author did an effort to explore the “Impact of artificial light intensity on nocturnal insects found in urban and rural areas of the Asir province, Saudi Arabia”. The subject is really an interesting to read because the growing urbanization is adversely damaging the ecosystem of entire life in the world including insects. Insects are very beneficial for the life security as their contribution in crop pollination, honey production, and predator’s pf several other insect species are very important for the agriculture sector and food availability. In the present study author, collected several insect species form the rural and urban areas in the Asir province of Saudi Arabia by using the latest techniques of insect collection. Present work highlights the importance of urban ecology study with reference to insect population and biodiversity in general. The presentation of the paper especially the introduction and discussion part is further improved. REVIEWER-3 1 Abstract Line 19: Change“street light columns” by “street-light columns” Line 23: Change“Light intensity” by “The light intensity” Line 24: Change“light meter” by “a light meter” Line 25: Change“darkness the sky” by “dark the sky” Done 2 Introduction Line 36: Change“Decline in insect” by “A decline in insect” Line 49: Change“new phenomenal” by “new phenomenon” Line 60: Change“decreasing number” by “the decreasing number” Line 60: Change“maintenance” by “the maintenance” Line 67: Change“Arabia the effect” by “Arabia, the effect” Line 68: Change“ALAN on insect” by “ALAN on the insect” Done 3 Materials and methods Line 81: Change“An agricultural farm land” by “Agricultural farmland” Line 104: Change“attracts and captures” by “attract and capture” Line 116: Change“boxplot” by “the boxplot” Done 4 Results Line 157: Change“high pressure sodium” by “high-pressure sodium” Line 157: Change“installed in many” by “installed on many” Line 158: Change“increasing in an alarming” by “increasing at an alarming” Done 5 Discussion Line 173: Change“maintain healthy” by “maintain a healthy” Line 173: Change“balanced diversity” by “a balanced diversity” Line 177: Change“of population” by “of the population” Line 184: Change“the possible reason” by “a possible reason” Line 188: Change“fauna including” by “fauna, including” Line 194: Change“in an agricultural farm land” by “in agricultural farmland” Line 196: Change“have disruptive” by “have a disruptive Done 6 Conclusions Line 200: Change“affect insect” by “affect the insect” Line 202: Change“ Warm light” by “The warm light” Done 7 On the other hand, there is repetition of figure 5 and table 2, therefore use one thing. Done 8 Moreover , following latest reference could be cited Cited Yours truly, 19-October-2020 Khalid Ali Khan, Ph.D. (Corresponding author) Unit of Bee Research and Honey Production, Biology Department, Faculty of Science, King Khalid University Abha 61413, Saudi Arabia 10.1371/journal.pone.0242315.r003 Decision Letter 1 Farooq Shahid Academic Editor © 2020 Shahid Farooq2020Shahid FarooqThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Submission Version1 29 Oct 2020 PONE-D-20-30451R1 Impact of artificial light intensity on nocturnal insect diversity in urban and rural areas of the Asir province, Saudi Arabia PLOS ONE Dear Dr. Khan, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== ACADEMIC EDITOR: I have just gone through the files uploaded by the authors. Unfortunately, the track-changed manuscript and clean copy are different files, which made evaluation difficult. Therefore, I request authors to please upload same file for track-changed and clean copies. ============================== Please submit your revised manuscript by Dec 13 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Shahid Farooq, Ph.D. Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. 10.1371/journal.pone.0242315.r004 Author response to Decision Letter 1 Submission Version2 29 Oct 2020 Dear Dr. Shahid Farooq. Academic Editor PLOS ONE Subject: SUBMISSION OF REVISED MANUSCRIPT (PONE-D-20-30451) Thank you very much for your comments regarding our manuscript titled: “Impact of artificial light intensity on nocturnal insects found in urban and rural areas of the Asir province, Saudi Arabia”. I have gone through all the changes suggested by the academic editor and reviewers. They pointed out very important points and provided very useful suggestions. I gave revised the manuscript accordingly. I have accepted all the changes suggested by the reviewers. Comments and responses of all the critique are presented in revised manuscript. The detail of responses to the editor/ reviewer’s comments is provided below in a tabular form. ACADEMIC EDITOR 1 The site selection needs clarity. One which basis these sites were selected? The delimitation of the site selection is provided in the text 2 Figure 1 must be replaced with a better one. Replaced 3 Introduction must be rearranged as suggested by the reviewers. Rearranged accordingly. 4 There should be more reference citations in the discussion section to support your claims. Done. Latest reference included 5 In acknowledgment figure 2 is about sky quality meter instead of insects. It is corrected and now this figure 2 replaced with figure 5. 6 Can the scale of Figure 5B be changed? Scale of Fig.5B Changed from 50 to 15. Journal requirements 1 Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. Checked 2 We suggest you thoroughly copyedit your manuscript for language usage, spelling, and grammar. If you do not know anyone who can help you do this, you may wish to consider employing a professional scientific editing service The language of the manuscript has been edited by Dr. Philip Smith, Uludağ University, Turkey. The language editing certificate is attached as other file. 3 In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why? No specific permits were required to execute the study and the work did not involve any endangered species 4 We note that Figure 1 in your submission contains map images which may be copyrighted. The map was created by R statistical environment with open source shape files. Therefore, the figure is not copyrighted. The detailed explanation is provided in the legend of Figure 1. REVIEWER-1 1 Introduction: T Introduction is a good review of the knowledge on the subject. However , the presentation of the light impact behavior with different factors involved like declining insect population, impact of artificial light, local impact etc., need to be improved/ arranged paragraph wise. Also, the introductory part should be more elaborative with latest references in order to provide more comprehensive presentation of the problem. Recent reports of insect population declines might be representing a global phenomenon in which the many stressors of the Anthropocene are pushing insects over the edge of population viability. This interesting aspect of artificial light use is defiantly stressing insects, we need to understand such patterns on insect decline and mechanisms. the authors must provide for these assertions/assumptions while writing introduction. The introduction is thoroughly rearranging with the additions of latest reference as per the suggestion of referee. 2 Methodology: The methodology is succinct presented but correct. The figures are satisfactory, except Figure 1 which is poorly structured. The main purpose of these figures is to show the localization of rural and urban region of study area. Author must replace this with better one. I guess the work is very limited to the number of locations and even why this location is specifically chosen is not well expressed. Also the number of samples replicates have not been mentioned. The devices used to perform light intensity data and insect collection traps are standard one, I did not find the methods of insect identification used in this paper . Methodology is sufficient and the query of reviewer regarding the site delimitation is added in the text. The number of samples replicates have already mentioned in text Insect identification part is corrected in MS with appropriate references. 3 Results: It is the best part of the manuscript. The results are focused and well presented on the insect population dynamics from both rural and urban areas. The section is further improved as suggested by reviewer-1 4 Discussion: The discussion consists in a good comparison of the insect populations from both urban and rural areas. However, there is a fewer reference in the discussion section. The authors must provide more recent citations to justify the current study with previous one This section also rearranged as per the advice of reviewer and also the latest references have been provided in the justification of the present study REVIEWER-2 1 In the present manuscript the author did an effort to explore the “Impact of artificial light intensity on nocturnal insects found in urban and rural areas of the Asir province, Saudi Arabia”. The subject is really an interesting to read because the growing urbanization is adversely damaging the ecosystem of entire life in the world including insects. Insects are very beneficial for the life security as their contribution in crop pollination, honey production, and predator’s pf several other insect species are very important for the agriculture sector and food availability. In the present study author, collected several insect species form the rural and urban areas in the Asir province of Saudi Arabia by using the latest techniques of insect collection. Present work highlights the importance of urban ecology study with reference to insect population and biodiversity in general. The presentation of the paper especially the introduction and discussion part is further improved. REVIEWER-3 1 Abstract Line 19: Change“street light columns” by “street-light columns” Line 23: Change“Light intensity” by “The light intensity” Line 24: Change“light meter” by “a light meter” Line 25: Change“darkness the sky” by “dark the sky” Done 2 Introduction Line 36: Change“Decline in insect” by “A decline in insect” Line 49: Change“new phenomenal” by “new phenomenon” Line 60: Change“decreasing number” by “the decreasing number” Line 60: Change“maintenance” by “the maintenance” Line 67: Change“Arabia the effect” by “Arabia, the effect” Line 68: Change“ALAN on insect” by “ALAN on the insect” Done 3 Materials and methods Line 81: Change“An agricultural farm land” by “Agricultural farmland” Line 104: Change“attracts and captures” by “attract and capture” Line 116: Change“boxplot” by “the boxplot” Done 4 Results Line 157: Change“high pressure sodium” by “high-pressure sodium” Line 157: Change“installed in many” by “installed on many” Line 158: Change“increasing in an alarming” by “increasing at an alarming” Done 5 Discussion Line 173: Change“maintain healthy” by “maintain a healthy” Line 173: Change“balanced diversity” by “a balanced diversity” Line 177: Change“of population” by “of the population” Line 184: Change“the possible reason” by “a possible reason” Line 188: Change“fauna including” by “fauna, including” Line 194: Change“in an agricultural farm land” by “in agricultural farmland” Line 196: Change“have disruptive” by “have a disruptive Done 6 Conclusions Line 200: Change“affect insect” by “affect the insect” Line 202: Change“ Warm light” by “The warm light” Done 7 On the other hand, there is repetition of figure 5 and table 2, therefore use one thing. Done 8 Moreover , following latest reference could be cited Cited Yours truly, 19-October-2020 Khalid Ali Khan, Ph.D. (Corresponding author) Unit of Bee Research and Honey Production, Biology Department, Faculty of Science, King Khalid University Abha 61413, Saudi Arabia 10.1371/journal.pone.0242315.r005 Decision Letter 2 Farooq Shahid Academic Editor © 2020 Shahid Farooq2020Shahid FarooqThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Submission Version2 2 Nov 2020 Impact of artificial light intensity on nocturnal insect diversity in urban and rural areas of the Asir province, Saudi Arabia PONE-D-20-30451R2 Dear Dr. Khan, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Shahid Farooq, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): I have evaluated the revised manuscript. All the queries raised by the referees have been properly addressed. Therefore, the current version is accepted for publication. Reviewers' comments: 10.1371/journal.pone.0242315.r006 Acceptance letter Farooq Shahid Academic Editor © 2020 Shahid Farooq2020Shahid FarooqThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 19 Nov 2020 PONE-D-20-30451R2 Impact of artificial light intensity on nocturnal insect diversity in urban and rural areas of the Asir province, Saudi Arabia Dear Dr. Khan: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Shahid Farooq Academic Editor PLOS ONE ==== Refs References 1 Sánchez-Bayo F , Wyckhuys KA . Worldwide decline of the entomofauna: A review of its drivers . Biological conservation . 2019 ;232 :8 –27 . 2 Grubisic M , van Grunsven RH , Kyba CC , Manfrin A , Hölker F . Insect declines and agroecosystems: does light pollution matter ? Annals of applied biology . 2018 ;173 (2 ):180 –9 . 3 Hallmann C , Sorg M , Jongejans E , Siepel H , Hofland N , Schwan H , et al More than 75 percent decline over 27 years in total flying insect biomass in protected areas . 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