Next Article in Journal
Phylogeny and Biogeographic History of Parnassius Butterflies (Papilionidae: Parnassiinae) Reveal Their Origin and Deep Diversification in West China
Next Article in Special Issue
Synanthropic Flies—A Review Including How They Obtain Nutrients, along with Pathogens, Store Them in the Crop and Mechanisms of Transmission
Previous Article in Journal
Monitoring Trends in Distribution and Seasonality of Medically Important Ticks in North America Using Online Crowdsourced Records from iNaturalist
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Insects
Volume 13
Issue 5
10.3390/insects13050405
insects-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Wing Phenotypic Variation among Stomoxys calcitrans (Diptera: Muscidae) Populations in Thailand

by
Tanawat Chaiphongpachara
1,
Gerard Duvallet
2 and
Tanasak Changbunjong
3,4,*
1
Department of Public Health and Health Promotion, College of Allied Health Sciences, Suan Sunandha Rajabhat University, Bangkok 10300, Thailand
2
UMR5175, Université Paul-Valéry Montpellier, 34090 Montpellier, France
3
Department of Pre-Clinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand
4
The Monitoring and Surveillance Center for Zoonotic Diseases in Wildlife and Exotic Animals (MoZWE), Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand
*
Author to whom correspondence should be addressed.
Insects 2022, 13(5), 405; https://doi.org/10.3390/insects13050405
Submission received: 23 March 2022 / Revised: 6 April 2022 / Accepted: 21 April 2022 / Published: 23 April 2022
(This article belongs to the Special Issue Biology, Ecology and Control of Synanthropic Flies)
Browse Figures
Versions Notes

Abstract

:

Simple Summary

The stable fly, Stomoxys calcitrans (Diptera: Muscidae), is the predominant Stomoxys spp. in Thailand and is considered a pest for livestock, pets, wildlife, and occasionally humans. This study investigated the phenotypic variation in the wing size and shape of S. calcitrans populations from different geographical regions in Thailand using landmark-based geometric morphometric analysis. Results showed strong spatial variation in wing shape among S. calcitrans populations and thus suggested the existence of phenotypic plasticity in this fly.

Abstract

Stomoxys calcitrans (Linnaeus, 1758) (Diptera: Muscidae) is a cosmopolitan hematophagous ectoparasite of veterinary and medical importance. It is an important mechanical vector of several animal pathogens and can cause significant economic losses. However, the morphological variation of this species remains unknown. This study aimed to investigate the phenotypic variation in the wing size and shape of S. calcitrans populations in Thailand based on a landmark-based geometric morphometric approach. Specimens were collected from five populations in five geographical regions in Thailand. A total of 490 left wings of S. calcitrans (245 female and 245 male individuals) were used for geometric morphometric analysis. Wing size differences were detected between some populations of S. calcitrans, whereas wing shape differences were found among populations. Therefore, the phenotypic variation in S. calcitrans populations indicated that these populations are adaptive responses to local environmental pressures, suggesting the presence of phenotypic plasticity in this species.

1. Introduction

The stable fly, Stomoxys calcitrans (Linnaeus, 1758) (Diptera: Muscidae), is one of the 18 species within the subfamily Stomoxyinae and genus Stomoxys [1]. This fly is widely distributed worldwide, causing serious health problems to animals, especially livestock, but occasionally humans [2]. Both sexes of S. calcitrans are hematophagous and recognized as a mechanical vector of several animal pathogens, including viruses (i.e., equine infectious anemia virus, African swine fever virus, African horse sickness virus, bovine leukemia virus, bovine herpes virus, bluetongue virus, and lumpy skin disease virus [2,3,4,5,6]), protozoa (i.e., Trypanosoma spp. and Besnoitia besnoiti [7,8]), bacteria (i.e., Bacillus anthracis [9] and Anaplasma marginale [2]), and helminths (i.e., Habronema microstoma [10]). Since S. calcitrans can transmit many pathogens, understanding its biology is very important as key knowledge for effective surveillance and control [11].
Phenotypic plasticity, which is also called phenotypic responsiveness, is the capacity of an organism to exhibit distinct phenotypes in response to stimuli from different environments [12]. Environmental heterogeneity affects phenotypic patterns, and these changes can increase the organism’s fitness [13]. Previous studies reported that measurable phenotypic variation could represent the phenotypic plasticity of insect vectors [12,14,15,16]. Currently, wing size and shape are valuable indicators for evaluating the phenotypic variation of insects to adapt to spatially different environments [13,15,17]. Furthermore, variation of the wings of insect vectors is also related to host-feeding sources and flight [12]. In fact, the change in the wing shape of insects can affect their flight capacity, whereas the change in the wing size can be used to estimate the change in body size [18,19]. The longevity of insect vectors has the strongest influence on vectorial capacity due to increased chances of pathogen transmission [20,21]. Barreaux et al. [21] reported the relation between size and longevity in some environments of malaria vector Anopheles gambiae (Diptera: Culicidae). Furthermore, Costanzo et al. [22] found the size-fecundity relationship of Aedes albopictus (Diptera: Culicidae) when their larvae were reared at high temperatures and low resource levels. Recently, Baleba et al. [23] reported that larval density and substrate quality affected the wing size and shape of S. calcitrans and the physical change in wings could significantly affect their flight and dispersion.
Thailand is a Southeast Asian country with a tropical climate [24]. Each geographical region has a varied topography and biodiversity of organisms [24,25]. There are six species of Stomoxys spp. distributed in Thailand, but S. calcitrans is the most predominant species [26,27]. Muenworn et al. [26] surveyed the distribution of stable flies within six geographical regions in Thailand and indicated that environmental conditions in each collection site were related to the density of flies. However, morphological variation in the wing size and shape of S. calcitrans in each geographical region of Thailand remains unknown. The lack of knowledge about insect phenotypic plasticity can be a major obstacle to species identification. Investigators are hesitant about specimens with unusual characteristics, leading to ineffective control measures [14,28].
Geometric morphometrics is a valuable tool and is becoming popular in the study of insect vectors in evaluating size and shape variations. The correlation between size and shape is known as allometry [14]. In insects, wings are the preferred structure for morphometric analyses due to their two-dimensional configurations reducing digitizing error [14]. Geometric morphometric analysis can be performed using three methods: landmark, semi-landmark, and outline-based [14,29,30]. The landmark-based method is most widely used for insect vector species to distinguish morphological closely related species [31,32], identify species [14,32], examine the phenotypic variation among populations [14,33,34], and determine sexual dimorphism [35]. Therefore, this study aimed to estimate the phenotypic variation in the wing size and shape of S. calcitrans among five populations in different geographical regions in Thailand based on a landmark-based geometric morphometric approach.

2. Materials and Methods

2.1. Ethical Statement

All study protocols were conducted according to the guidelines for biomedical research involving animals. This study was approved and endorsed by the Faculty of Veterinary Science, Mahidol University Animal Care and Use Committee (ethical approval no. MU-IACUC 2018/008).

2.2. Stable Fly Specimens

Male and female S. calcitrans specimens were collected from five populations (also called collection sites) representing five different geographical regions in Thailand, i.e., Mae Hong Son Province (MH; northern region), Nakhon Ratchasima Province (NR; northeastern region), Nakhon Pathom Province (NP; central region), Kanchanaburi Province (KB; western region), and Songkhla Province (SK; southern region; Table 1; Figure 1). Five Nzi traps [36] were used to collect S. calcitrans from each population for 2 consecutive days (from 06:00 to 18:00) between February and July 2018. The traps were made locally, using blue and black fabric named Solon® (Bangkok, Thailand) being 100% polyester. The specimens were collected at 2 or 3 h intervals to prevent specimen damage for morphological identification. All specimens were immediately euthanized by freezing at −10 °C, individually placed in 1.5 mL microcentrifuge tubes, and sent to the Vector-Borne Diseases Research Unit, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand. The species were identified based on morphological characters by the taxonomic keys of Zumpt [1] and Tumrasvin and Shinonaga [37] under a stereomicroscope (Nikon SMZ745; Nikon Corp., Tokyo, Japan). The specimens were stored at −20 °C until they were used for geometric morphometric analysis.

2.3. Sample Preparation and Landmark Digitization

The left wings of male and female S. calcitrans were detached from the thorax using a sterilized blade and mounted between a microscope slide and cover glass with Hoyer’s medium [31]. Each mounted wing slide was photographed using a digital camera coupled to a stereomicroscope (Nikon AZ 100; Nikon) at 10× magnification and embedded in all wing images at a 1 mm scale unit. Then, ten anatomical landmarks on the intersections of wing veins and intersection with the wing borders [31] were digitized (Figure 2) using XYOM (XY Online Morphometrics) version 2 software [30], which was freely accessed at https://xyom.io/me, accessed on 15 February 2022. The geometric and statistical analyses and graphic outputs were also performed by XYOM.

2.4. Repeatability and Allometry

Before wing size and shape analyses, repeatability and allometry were examined. A repeatability test is important to assess the accuracy of landmark digitization based on comparing two sets of wing images. Ten images per population of male and female S. calcitrans were randomly selected and digitized twice by the same user. The repeatability index was computed based on the Procrustes analysis of variance (ANOVA) method to examine the measurement error of landmark digitization [38]. As for allometry, the estimation of the allometric effect is also important to assess the effect of the wing size on wing shape variation. Linear regression based on the first (shape-derived) discriminant factor (DF) on wing size was used in this investigation and estimated by the determination coefficient (r2).

2.5. Wing Size Analyses

Centroid size (CS) was calculated from the square root of the sum of squared distances between the centroid and each landmark to represent the global wing size of S. calcitrans to determine size variation among different populations [39]. Graphic quantile boxes were built to display wing CS variations of S. calcitrans in each population. Differences in the average wing CS of male and female S. calcitrans between populations were compared using one-way ANOVA followed by Bonferroni post hoc test. A nonparametric procedure (1000 permutations) was used to estimate statistical significance at p < 0.05.

2.6. Wing Shape Analyses

The wing shape variables were obtained through a Procrustes superimposition according to the Generalized Procrustes Analysis. The principal components of shape variables were used as final shape variables for wing shape analysis. Wing shape variations of male and female S. calcitrans among populations were estimated by discriminant analysis according to the first two DFs and illustrated by factor maps. The Mahalanobis distance was calculated to estimate the metric distance of shape divergence between groups. The statistical significance of average wing shape differences based on Mahalanobis distances of male and female S. calcitrans between populations was calculated by a nonparametric permutation test (1000 permutations) at p < 0.05. Furthermore, to assess the relationships of wing shape of S. calcitrans among populations, a UPGMA algorithm based on the Mahalanobis distances was used to illustrate a hierarchical clustering tree. Branch support was estimated based on 1000 bootstrap replicates for each data set.

2.7. Validated Classification

A cross-validated classification test was used to analyze the accuracy of wing size and shape that may be specific to different populations. Each individual sample was sequentially removed from the total sample and assigned to the most likely (for size) and closest group (for shape) based on the maximum likelihood method and Mahalanobis distance, respectively.

3. Results

In this study, 490 S. calcitrans (245 female and 245 male individuals) collected from five populations representing five geographical regions in Thailand were used to examine the phenotypic variation of wing size and shape using the landmark-based geometric morphometric method.

3.1. Repeatability of Wing Image

The quality in digitizing landmarks of our wing image set based on testing repeatability revealed that the repeatability score of male and female S. calcitrans was high (95% for shape). Meanwhile, the measurement error was low (5% for shape).

3.2. Allometric Effect

An analysis of the relationship between the size and shape of male and female S. calcitrans showed that wing size variation was significantly correlated to wing shape changes (p < 0.05; Figure 3). Linear regression prediction revealed a negative correlation, meaning a smaller wing size correlated with a greater difference in wing shape (r2 = 15.0% for females and r2 = 24.0% for males).

3.3. Wing Size Variation

Overall, the wing CS of female S. calcitrans (ranged from 4.09 to 4.43 mm) was larger than that of males (ranged from 3.97 to 4.30 mm; Figure 4). The analysis of the wing CS variation of female S. calcitrans among different populations in Thailand showed that the MH population was the largest (4.43 mm), followed by NR and NP (4.40 mm), KB (4.34 mm), and SK (4.09 mm), whereas the wing CS variation of male S. calcitrans showed that the NP population was the largest (4.30 mm), followed by MH (4.29 mm), NR (4.23 mm), KB (4.14 mm), and SK (3.97 mm; Table 2). A statistically significant difference in the wing CS of female S. calcitrans was found between the SK population and all population groups (p < 0.05), whereas statistical significance in the wing CS of male S. calcitrans was found between the SK population and all population groups (p < 0.05), MH and KB (p < 0.05), and NP and KB (p < 0.05).

3.4. Wing Shape Variation

After the generalized Procrustes analysis, graphic constructions of the wing shape of female and male S. calcitrans were built from the superimposition of aligned mean configurations. These graphic wing constructions revealed the most visible displacement at 1, 2, 7, and 10 landmark positions (Figure 5).
The analysis of the wing shape variations among S. calcitrans populations in factor maps based on discriminant analysis defined by DF axes showed that the first two DF axes accounted for 89% of the total wing shape variation for female S. calcitrans (DF1 = 73% and DF2 = 16%) and 91% for male S. calcitrans (DF1 = 77% and DF2 = 14%; Figure 6). All female S. calcitrans populations represented overlapping, and no distinct populations were separated from each other. In contrast, all-male populations represented a majority overlap, and nearly all populations were not separated, except for the SK population separated from MH, NR, and NP (Figure 6). However, comparing pairwise Mahalanobis distances of male and female S. calcitrans between populations showed statistically significant differences (p < 0.05, Table 3). A hierarchical clustering tree based on Mahalanobis distances of S. calcitrans populations showed the same pattern based on the wing shape between females and males (Figure 7). The wing shape of MH was similar to NR than NP, whereas KB was similar to SK and separated from other populations supported by 100% bootstrap values. The tree also showed clear wing shape differences between females and males based on group separation.
Cross-validated classification yielded low and high correctly assigned scores for wing size (0–58%) and shape (51.11–80%), respectively (Table 4).

4. Discussion

The study of phenotypic variation is important to understand the influence of environmental and/or genetic factors in a population. This study investigated wing size and shape variations among S. calcitrans populations representing five different geographical regions in Thailand based on a geometric morphometric approach. The wing size of female and male S. calcitrans in SK was significantly smaller than in other populations. SK is a coastal province in Thailand located near the Gulf of Thailand. The influence of coastal climates may affect the wing size of S. calcitrans. Previous studies indicated that some insects in coastal areas have smaller wing sizes than those in mainland areas. Sumruayphol et al. [40] studied Phlebotomus stantoni (Diptera: Psychodidae) from different provinces in Thailand and found that the wing size of the population in Lang Ga Jiew Island, Chumphon Province, was the smallest. Demari-Silva et al. [41] found that Culex coronator (Diptera: Culicidae) populations in Brazil’s Rio de Janeiro Municipality lowland coastal areas had significantly smaller wing sizes than in other areas. However, some dipteran insects exhibited the opposite effect depending on their suitability for life in coastal areas. Chaiphongpachara et al. [17] found that the average wing size of female Aedes aegypti (Diptera: Culicidae) in coastal areas was significantly larger than in the residential and cultivated areas in Samut Songkhram Province, Thailand.
Furthermore, high-quality food sources and their population density in each area are key factors in the wing size changes of insects [42]. High food quality and suitable population density result in larger wing sizes of insects than in food-poor areas and high population density [23,42]. Consequently, different wing sizes in each area can account for the degree of suitability of their habitat [23]. In this study, the habitat of S. calcitrans in southern Thailand was a zoo located in an urban area. Changbunjong et al. [27] studied stomoxyine flies across Thailand and found that zoos have a denser S. calcitrans population than livestock farms. Although zoos have a wide variety of animal hosts, they have limited spatial restrictions and are located in urban areas, resulting in a low level of biodiversity of stomoxyine flies and making the S. calcitrans population the most predominant species in zoos without competing species [27].
Wing shape is related to flying capability [43]. This capacity of insect vectors is used for host-seeking. Data analyses revealed that the wing shape of female and male S. calcitrans indicates the variation between different populations of Thailand. Wing shape differences are caused by different environmental influences. Previous studies reported that certain environmental factors affect the insect wing shape [15,44,45]. Phanitchat et al. [46] studied the change in Ae. albopictus wings with temperature and found that wing shape changed with increasing temperature. In addition, larval density and the nature of the developmental substrate are also natural factors to influence the wing shape of S. calcitrans [23]. Meanwhile, the seasonal variations could have an impact on wing shape as well as wing size of S. calcitrans in our study. Prudhomme et al. [34] found that seasonal environments affect wing shape and size variations in Phlebotomus ariasi (Diptera: Psychodidae). However, environmental factors can influence wing size much more than wing shape [14].
A hierarchical clustering tree revealed the proximity of wing shape among S. calcitrans populations in Thailand. It was supposed that wing shape variation might be related to the altitude of the areas. The altitude causes different ecosystems due to different environmental factors, such as levels of sunlight, temperature, wind, relative humidity, host species, and resident plant species [47]. The wing shapes of northern (MH) and northeastern (NR) populations were similar. These results may be because both sites are located at high altitudes (452 and 498 m). The wing shapes of western (KB) and southern (SK) populations were similar. Both sites are moderate-altitude areas (174 and 112 m). The wing shape of the central (NP) population as a low-altitude area (11 m) was sandwiched between those populations. These results were consistent with Lorenz et al. [47] that wing shapes of the malaria vector Anopheles cruzii (Diptera: Culicidae) in Brazil were distinct between lowland (altitude of 5–20 m) and hilltop (altitude of 81–263 m) populations. The comparison of pairwise Mahalanobis distances in this study indicated that populations in southern regions had a different wing shape than other regions. The southern region of Thailand is the most different area from other regions. This region is along the Andaman Sea and the Gulf of Thailand and has only two seasons, summer and rainy, whereas other regions have three seasons, summer, rainy, and winter [48]. This phenomenon might explain the highest wing shape variation among populations of S. calcitrans in Thailand. The results from the hierarchical clustering tree also revealed a sexual dimorphism in the wing shape of S. calcitrans in Thailand. These results indicated that the phenotypic expression of wing shape was a sex-specific difference. These results were consistent with a previous report on other Stomoxys spp. (Stomoxys indicus (Diptera: Muscidae), Stomoxys pullus (Diptera: Muscidae), and Stomoxys uruma (Diptera: Muscidae)) [31]. The sexual dimorphism of wing shape was also reported in other insect vectors, such as mosquitoes of the genera Aedes (Ae. aegypti and Ae. albopictus), Anopheles (Anopheles albitarsis (Diptera: Culicidae), Anopheles cruzii (Diptera: Culicidae), Anopheles homunculus (Diptera: Culicidae), Anopheles strode (Diptera: Culicidae), and Anopheles triannulatus (Diptera: Culicidae)), Culex (Culex quinquefasciatus (Diptera: Culicidae) and Culex nigripalpus (Diptera: Culicidae)), and Ochlerotatus (Ochlerotatus scapularis (Diptera: Culicidae)) [49] and the biting fly Haematobosca aberrans (Diptera: Muscidae) [35]. Furthermore, sexual shape dimorphism in this study was correlated with sexual size dimorphism (female, 15%; male, 24%). This suggested that allometry is an important factor in sexual shape dimorphism in S. calcitrans wings.
In this study, cross-validated classification scores showed that the wing shape of S. calcitrans was more specific to each population than wing size (18.37% for size and 61.63% for shape). These results indicated that wing shapes of S. calcitrans expressed phenotypes in response to the unique topography of each geographical region more than wing size. Wing size is a very sensitive factor to the environment compared to wing shape [50]. Therefore, wing size was highly variable and did not reflect interregional specificity in this study. All results were supported by linear regression prediction of the relationship between size and shape based on allometry. Linear prediction explained that a smaller wing size correlated with a greater difference in wing shape. This relationship can explain the natural pressures from environmental differences. However, non-allometric effects, such as flight behavior and mating systems, may be related to variations in wing shape [51].
Although phenotypic variation results from environmental factors, it can also result from genetic factors [14]. However, a previous study on genetic differences based on cytochrome c oxidase subunit I in S. calcitrans populations from Thailand showed that low intraspecific divergence ranged from 0 to 3.2% (mean = 0.8%) [52]. In addition, a study of the population structure of S. calcitrans from nine provinces of Thailand using allele variation frequencies of isozymes demonstrated no significant genetic difference among the nine populations [53]. Hence, these results indicated that the phenotypic plasticity of the S. calcitrans population in this study might not be related to genetic variation. In contrast, genetic factors affected the phenotypic variation of some insect species, such as Drosophila melanogaster (Diptera: Drosophilidae) [54].

5. Conclusions

This study provided phenotypic information on S. calcitrans populations in Thailand based on a landmark-based geometric morphometric analysis of wing size and shape variations. Results showed that wing size differences were detected between some populations of S. calcitrans, whereas wing shape differences were detected among populations. Wing size variation had a significant effect on wing shape variation. These results indicated that phenotypic variation in S. calcitrans is an adaptive response to local environmental pressures in the study areas. It was speculated that the effects of this physical change might affect the biology of this species in the aspects of dispersion and spread of the diseases. However, the relationship between phenotypic variation and environmental factors and/or pathogen transmission should be considered in further studies.

Author Contributions

Conceptualization, T.C. (Tanawat Chaiphongpachara), T.C. (Tanasak Changbunjong) and G.D.; methodology, T.C. (Tanawat Chaiphongpachara) and T.C. (Tanasak Changbunjong); validation, T.C. (Tanawat Chaiphongpachara), T.C. (Tanasak Changbunjong) and G.D.; investigation, T.C. (Tanawat Chaiphongpachara) and T.C. (Tanasak Changbunjong); resources, T.C. (Tanasak Changbunjong); data curation, T.C. (Tanawat Chaiphongpachara) and T.C. (Tanasak Changbunjong); writing—original draft preparation, T.C. (Tanawat Chaiphongpachara) and T.C. (Tanasak Changbunjong); writing—review and editing, T.C. (Tanawat Chaiphongpachara), T.C. (Tanasak Changbunjong) and G.D.; project administration, T.C. (Tanasak Changbunjong); funding acquisition, T.C. (Tanasak Changbunjong). All authors have read and agreed to the published version of the manuscript.

Funding

This study was financially supported by the Faculty of Veterinary Science, Mahidol University.

Institutional Review Board Statement

This study was approved by the Faculty of Veterinary Science, Mahidol University Animal Care and Use Committee (Ref. MU-IACUC 2018/008).

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Acknowledgments

We would like to thank the authorities of the Monitoring and Surveillance Center for Zoonotic Diseases in Wildlife and Exotic Animals (MoZWE) for their kind cooperation and assistance in the study.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Zumpt, F. The Stomoxyine Biting Flies of the World; Gustav Fisher Verlag: Stuttgart, Germany, 1973; pp. 97–137. [Google Scholar]
  2. Baldacchino, F.; Muenworn, V.; Desquesnes, M.; Desoli, F.; Charoenviriyaphap, T.; Duvallet, G. Transmission of pathogens by Stomoxys flies (Diptera, Muscidae): A review. Parasite 2013, 20, 26. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  3. Gibbs, E.P.J.; Johnson, R.H.; Gatehouse, A.G. A laboratory technique for studying the mechanical transmission of bovine herpes mammillitis virus by the stable fly (Stomoxys calcitrans L.). Res. Vet. Sci. 1973, 14, 145–147. [Google Scholar] [PubMed]
  4. Foil, L.D.; Meek, C.L.; Adams, W.V.; Issel, C.J. Mechanical transmission of equine infectious anemia virus by deer flies (Chrysops flavidus) and stable flies (Stomoxys calcitrans). Am. J. Vet. Res. 1983, 44, 155–156. [Google Scholar] [PubMed]
  5. Sohier, C.; Haegeman, A.; Mostin, L.; De Leeuw, I.; Campe, W.V.; De Vleeschauwer, A.; Tuppurainen, E.S.M.; van den Berg, T.; De Regge, N.; De Clercq, K. Experimental evidence of mechanical lumpy skin disease virus transmission by Stomoxys calcitrans biting flies and Haematopota spp. horseflies. Sci. Rep. 2019, 9, 20076. [Google Scholar] [CrossRef]
  6. Vergne, T.; Andraud, M.; Bonnet, S.; De Regge, N.; Desquesnes, M.; Fite, J.; Etore, F.; Garigliany, M.M.; Jori, F.; Lempereur, L.; et al. Mechanical transmission of African swine fever virus by Stomoxys calcitrans: Insights from a mechanistic model. Transbound. Emerg. Dis. 2021, 68, 1541–1549. [Google Scholar] [CrossRef]
  7. Desquesnes, M.; Holzmuller, P.; Lai, D.H.; Dargantes, A.; Lun, Z.R.; Jittaplapong, S. Trypanosoma evansi and surra: A review and perspectives on origin, history, distribution, taxonomy, morphology, hosts, and pathogenic effects. Biomed. Res. Int. 2013, 2013, 194176. [Google Scholar] [CrossRef] [Green Version]
  8. Sharif, S.; Jacquiet, P.; Prevot, F.; Grisez, C.; Raymond-Letron, I.; Semin, M.O.; Geffré, A.; Trumel, C.; Franc, M.; Bouhsira, É.; et al. Stomoxys calcitrans, mechanical vector of virulent Besnoitia besnoiti from chronically infected cattle to susceptible rabbit. Med. Vet. Entomol. 2019, 33, 247–255. [Google Scholar] [CrossRef] [Green Version]
  9. Turell, M.J.; Knudson, G.B. Mechanical transmission of Bacillus anthracis by stable flies (Stomoxys calcitrans) and mosquitoes (Aedes aegypti and Aedes taeniorhynchus). Infect. Immun. 1987, 55, 1859–1861. [Google Scholar] [CrossRef] [Green Version]
  10. Traversa, D.; Otranto, D.; Iorio, R.; Carluccio, A.; Contri, A.; Paoletti, B.; Bartolini, R.; Giangaspero, A. Identification of the intermediate hosts of Habronema microstoma and Habronema muscae under field conditions. Med. Vet. Entomol. 2008, 22, 283–287. [Google Scholar] [CrossRef]
  11. Cook, D. A historical review of management options used against the stable fly (Diptera: Muscidae). Insects 2020, 11, 313. [Google Scholar] [CrossRef]
  12. Nattero, J.; Leonhard, G.; Gürtler, R.E.; Crocco, L.B. Evidence of selection on phenotypic plasticity and cost of plasticity in response to host-feeding sources in the major Chagas disease vector Triatoma infestans. Acta Trop. 2015, 152, 237–244. [Google Scholar] [CrossRef] [PubMed]
  13. Bai, Y.; Dong, J.J.; Guan, D.L.; Xie, J.Y.; Xu, S.Q. Geographic variation in wing size and shape of the grasshopper Trilophidia annulata (Orthoptera: Oedipodidae): Morphological trait variations follow an ecogeographical rule. Sci. Rep. 2016, 6, 32680. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  14. Dujardin, J.P. Morphometrics applied to medical entomology. Infect. Genet. Evol. 2008, 8, 875–890. [Google Scholar] [CrossRef]
  15. Alves, V.M.; Moura, M.O.; de Carvalho, C.J.B. Wing shape is influenced by environmental variability in Polietina orbitalis (Stein) (Diptera: Muscidae). Rev. Bras. Entomol. 2016, 60, 150–156. [Google Scholar] [CrossRef] [Green Version]
  16. Oguz, G.; Kasap, O.E.; Alten, B. Wing morphology variations in a natural population of Phlebotomus tobbi Adler and Theodor 1930. J. Vector Ecol. 2017, 42, 223–232. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  17. Chaiphongpachara, T.; Juijayen, N.; Chansukh, K.K. Wing geometry analysis of Aedes aegypti (Diptera, Culicidae), a dengue virus vector, from multiple geographical locations of Samut Songkhram, Thailand. J. Arthropod-Borne Dis. 2018, 12, 351–360. [Google Scholar] [PubMed]
  18. Gilchrist, A.S.; Partridge, L. The contrasting genetic architecture of wing size and shape in Drosophila melanogaster. Heredity 2001, 86, 144–152. [Google Scholar] [CrossRef] [Green Version]
  19. Dellicour, S.; Gerard, M.; Prunier, J.G.; Dewulf, A.; Kuhlmann, M.; Michez, D. Distribution and predictors of wing shape and size variability in three sister species of solitary bees. PLoS ONE 2017, 12, e0173109. [Google Scholar] [CrossRef]
  20. Barreaux, A.M.G.; Barreaux, P.; Thievent, K.; Koella, J.C. Larval environment influences vector competence of the malaria mosquito Anopheles gambiae. Malar. World J. 2016, 7, 1–6. [Google Scholar]
  21. Barreaux, A.M.G.; Stone, C.M.; Barreaux, P.; Koella, J.C. The relationship between size and longevity of the malaria vector Anopheles gambiae (s.s.) depends on the larval environment. Parasit. Vectors 2018, 11, 485. [Google Scholar] [CrossRef] [Green Version]
  22. Costanzo, K.S.; Westby, K.M.; Medley, K.A. Genetic and environmental influences on the size-fecundity relationship in Aedes albopictus (Diptera: Culicidae): Impacts on population growth estimates? PLoS ONE 2018, 13, e0201465. [Google Scholar] [CrossRef] [PubMed]
  23. Baleba, S.B.S.; Masiga, D.; Torto, B.; Weldon, C.W.; Getahun, M.N. Effect of larval density and substrate quality on the wing geometry of Stomoxys calcitrans L. (Diptera: Muscidae). Parasit. Vectors 2019, 12, 222. [Google Scholar] [CrossRef] [PubMed]
  24. Baimai, V. Biodiversity in Thailand. J. R. Inst. Thail. 2010, 2, 107–114. [Google Scholar]
  25. Kermel-Torres, D. Atlas of Thailand: Spatial Structures and Development; IRD Editions: Paris, France, 2004; pp. 13–33. [Google Scholar]
  26. Muenworn, V.; Duvallet, G.; Thainchum, K.; Tuntakom, S.; Tanasilchayakul, S.; Prabaripai, A.; Akratanakul, P.; Sukonthabhirom, S.; Chareonviriyaphap, T. Geographic distribution of stomoxyine flies (Diptera: Muscidae) and diurnal activity of Stomoxys calcitrans in Thailand. J. Med. Entomol. 2010, 47, 791–797. [Google Scholar] [CrossRef]
  27. Changbunjong, T.; Weluwanarak, T.; Ratanakorn, P.; Maneeon, P.; Ganpanakngan, M.; Apiwathnasorn, C.; Sungvornyothin, S.; Sriwichai, P.; Sumruayphol, S.; Ruangsittichai, J. Distribution and abundance of Stomoxyini flies (Diptera: Muscidae) in Thailand. Southeast Asian J. Trop. Med. Public Health 2012, 43, 1400–1410. [Google Scholar]
  28. Gibert, P.; Debat, V.; Ghalambor, C.K. Phenotypic plasticity, global change, and the speed of adaptive evolution. Curr. Opin. Insect Sci. 2019, 35, 34–40. [Google Scholar] [CrossRef]
  29. Dujardin, J.P.; Kaba, D.; Solano, P.; Dupraz, M.; McCoy, K.D.; Jaramillo-O, N. Outline-based morphometrics, an overlooked method in arthropod studies? Infect. Genet. Evol. 2014, 28, 704–714. [Google Scholar] [CrossRef]
  30. Dujardin, S.; Dujardin, J.P. Geometric morphometrics in the cloud. Infect. Genet. Evol. 2019, 70, 189–196. [Google Scholar] [CrossRef]
  31. Changbunjong, T.; Sumruayphol, S.; Weluwanarak, T.; Ruangsittichai, J.; Dujardin, J.P. Landmark and outline-based geometric morphometrics analysis of three Stomoxys flies (Diptera: Muscidae). Folia Parasitol. 2016, 63, 37. [Google Scholar] [CrossRef] [Green Version]
  32. Changbunjong, T.; Prakaikowit, N.; Maneephan, P.; Kaewwiset, T.; Weluwanarak, T.; Chaiphongpachara, T.; Dujardin, J.P. Landmark data to distinguish and identify morphologically close Tabanus spp. (Diptera: Tabanidae). Insects 2021, 12, 974. [Google Scholar] [CrossRef]
  33. Morales Vargas, R.E.; Phumala-Morales, N.; Tsunoda, T.; Apiwathnasorn, C.; Dujardin, J.P. The phenetic structure of Aedes albopictus. Infect. Genet. Evol. 2013, 13, 242–251. [Google Scholar] [CrossRef] [PubMed]
  34. Prudhomme, J.; Cassan, C.; Hide, M.; Toty, C.; Rahola, N.; Vergnes, B.; Dujardin, J.P.; Alten, B.; Sereno, D.; Bañuls, A.L. Ecology and morphological variations in wings of Phlebotomus ariasi (Diptera: Psychodidae) in the region of Roquedur (Gard, France): A geometric morphometrics approach. Parasit. Vectors 2016, 14, 578. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  35. Changbunjong, T.; Ruangsittichai, J.; Duvallet, G.; Pont, A.C. Molecular identification and geometric morphometric analysis of Haematobosca aberrans (Diptera: Muscidae). Insects 2020, 16, 451. [Google Scholar] [CrossRef] [PubMed]
  36. Mihok, S. The development of a multipurpose trap (the Nzi) for tsetse and other biting flies. Bull. Entomol. Res. 2002, 92, 385–403. [Google Scholar] [CrossRef]
  37. Tumrasvin, W.; Shinonaga, S. Studies on medically important flies in Thailand. V. On 32 species belonging to the subfamilies Muscinae and Stomoxyini including the taxonomic keys (Diptera: Muscidae). Bull. Tokyo Med. Dent. Univ. 1978, 25, 201–227. [Google Scholar]
  38. Arnqvist, G.; Martensson, T. Measurement error in geometric morphometrics: Empirical strategies to assess and reduce its impact on measures of shape. Acta Zool. Academ. Sci. Hung. 1998, 44, 73–96. [Google Scholar]
  39. Bookstein, F.L. Morphometric Tools for Landmark Data. Geometry and Biology; Cambridge University Press: Cambridge, UK, 1991. [Google Scholar]
  40. Sumruayphol, S.; Chittsamart, B.; Polseela, R.; Sriwichai, P.; Samung, Y.; Apiwathnasorn, C.; Dujardin, J.P. Wing geometry of Phlebotomus stantoni and Sergentomyia hodgsoni from different geographical locations in Thailand. C. R. Biol. 2017, 340, 37–46. [Google Scholar] [CrossRef]
  41. Demari-Silva, B.; Suesdek, L.; Sallum, M.A.M.; Marrelli, M.T. Wing geometry of Culex coronator (Diptera: Culicidae) from South and Southeast Brazil. Parasit. Vectors 2014, 7, 174. [Google Scholar] [CrossRef] [Green Version]
  42. Lin, X.; Xu, Y.; Jiang, J.; Lavine, M.; Lavine, L.C. Host quality induces phenotypic plasticity in a wing polyphenic insect. Proc. Natl. Acad. Sci. USA 2018, 115, 7563–7568. [Google Scholar] [CrossRef] [Green Version]
  43. Johansson, F.; Söderquist, M.; Bokma, F. Insect wing shape evolution: Independent effects of migratory and mate guarding flight on dragonfly wings. Biol. J. Linn. Soc. 2009, 97, 362–372. [Google Scholar] [CrossRef] [Green Version]
  44. Chaiphongpachara, T.; Laojun, S. Wing morphometric variability of the malaria vector Anopheles (Cellia) epiroticus Linton et Harbach (Diptera: Culicidae) for the duration of the rainy season in coastal areas of Samut Songkhram, Thailand. Folia Parasitol. 2020, 67, 1–7. [Google Scholar] [CrossRef] [PubMed]
  45. Morales Vargas, R.; Tsunoda, T.; Noda, J.; Bousses, P.; Nguyen, T.Y.; Hasebe, F.; Dujardin, J.P. Shape relatedness between geographic populations of Culex tritaeniorhynchus, the primary vector of Japanese encephalitis virus: A landmark study. Infect. Genet. Evol. 2021, 90, 104764. [Google Scholar] [CrossRef] [PubMed]
  46. Phanitchat, T.; Apiwathnasorn, C.; Sungvornyothin, S.; Samung, Y.; Dujardin, S.; Dujardin, J.P.; Sumruayphol, S. Geometric morphometric analysis of the effect of temperature on wing size and shape in Aedes albopictus. Med. Vet. Entomol. 2019, 33, 476–484. [Google Scholar] [CrossRef] [PubMed]
  47. Lorenz, C.; Marques, T.C.; Sallum, M.A.M.; Suesdek, L. Altitudinal population structure and microevolution of the malaria vector Anopheles cruzii (Diptera: Culicidae). Parasit. Vectors 2014, 7, 581. [Google Scholar] [CrossRef] [PubMed]
  48. Marks, D. Climate change and Thailand: Impact and response. Contemp. Southeast Asia 2011, 33, 229–258. [Google Scholar] [CrossRef]
  49. Virginio, F.; Oliveira Vidal, P.; Suesdek, L. Wing sexual dimorphism of pathogen-vector culicids. Parasit. Vectors 2015, 8, 159. [Google Scholar] [CrossRef] [Green Version]
  50. Lorenz, C.; Almeida, F.; Almeida-Lopes, F.; Louise, C.; Pereira, S.N.; Petersen, V.; Vidal, P.O.; Virginio, F.; Suesdek, L. Geometric morphometrics in mosquitoes: What has been measured? Infect. Genet. Evol. 2017, 54, 205–215. [Google Scholar] [CrossRef]
  51. Gidaszewski, N.A.; Baylac, M.; Klingenberg, C.P. Evolution of sexual dimorphism of wing shape in the Drosophila melanogaster subgroup. BMC Evol. Biol. 2009, 9, 110. [Google Scholar] [CrossRef] [Green Version]
  52. Changbunjong, T.; Weluwanarak, T.; Samung, Y.; Ruangsittichai, J. Molecular identification and genetic variation of stomoxyine flies (Diptera: Muscidae) in Thailand based on cox1 barcode. J. Asia Pac. Entomol. 2016, 19, 1117–1123. [Google Scholar] [CrossRef]
  53. Tainchum, K.; Sukonthabhirom, S.; Duvallet, G.; Akratanakul, P.; Muenworn, V.; Chareonviriyaphap, T. Population structure of Stomoxys calcitrans (Diptera: Muscidae) from nine regions of Thailand. J. Econ. Entomol. 2010, 103, 1012–1018. [Google Scholar] [CrossRef]
  54. Long, A.; Mullaney, S.; Reid, L.; Fry, J.; Langley, C.; Mackay, T.F.C. High resolution mapping of genetic factors affecting abdominal bristle number in Drosophila melanogaster. Genetics 1995, 139, 1273–1291. [Google Scholar] [CrossRef] [PubMed]
Insects 13 00405 g001 550
Figure 1. Map of S. calcitrans populations in Thailand: Mae Hong Son (1), Nakhon Ratchasima (2), Nakhon Pathom (3), Kanchanaburi (4), and Songkhla (5).
Figure 1. Map of S. calcitrans populations in Thailand: Mae Hong Son (1), Nakhon Ratchasima (2), Nakhon Pathom (3), Kanchanaburi (4), and Songkhla (5).
Insects 13 00405 g001
Insects 13 00405 g002 550
Figure 2. Ten anatomical landmarks selected from the left wing to estimate the phenotypic variation of S. calcitrans based on landmark-based geometric morphometric analysis.
Figure 2. Ten anatomical landmarks selected from the left wing to estimate the phenotypic variation of S. calcitrans based on landmark-based geometric morphometric analysis.
Insects 13 00405 g002
Insects 13 00405 g003 550
Figure 3. Linear regression between wing size (CS) and shape (DF) of female (top) and male (bottom) S. calcitrans. Orange dotted lines indicate linear regression prediction.
Figure 3. Linear regression between wing size (CS) and shape (DF) of female (top) and male (bottom) S. calcitrans. Orange dotted lines indicate linear regression prediction.
Insects 13 00405 g003
Insects 13 00405 g004 550
Figure 4. Quantile boxes of wing CS variations of male and female S. calcitrans populations. The horizontal line crossing each box is the median separating the 25th and 75th quartiles.
Figure 4. Quantile boxes of wing CS variations of male and female S. calcitrans populations. The horizontal line crossing each box is the median separating the 25th and 75th quartiles.
Insects 13 00405 g004
Insects 13 00405 g005 550
Figure 5. Superposition of the aligned mean anatomical landmark positions of female (top) and male (bottom) S. calcitrans populations. Enlarged images in small frames showed the parts of wing construction where variation occurred.
Figure 5. Superposition of the aligned mean anatomical landmark positions of female (top) and male (bottom) S. calcitrans populations. Enlarged images in small frames showed the parts of wing construction where variation occurred.
Insects 13 00405 g005
Insects 13 00405 g006 550
Figure 6. Factor maps based on discriminant analysis in wing shape variation of female (top) and male (bottom) S. calcitrans populations. Each point in a polygon represents an individual wing sample, and small squares in a polygon represent the position of the mean group. The horizontal axis was the first DF (DF1), whereas the vertical axis was the second DF (DF2).
Figure 6. Factor maps based on discriminant analysis in wing shape variation of female (top) and male (bottom) S. calcitrans populations. Each point in a polygon represents an individual wing sample, and small squares in a polygon represent the position of the mean group. The horizontal axis was the first DF (DF1), whereas the vertical axis was the second DF (DF2).
Insects 13 00405 g006
Insects 13 00405 g007 550
Figure 7. Hierarchical clustering tree based on Mahalanobis distances of female and male S. calcitrans populations. Numbers at the nodes indicate the percentages of bootstrap values based on 1000 bootstraps.
Figure 7. Hierarchical clustering tree based on Mahalanobis distances of female and male S. calcitrans populations. Numbers at the nodes indicate the percentages of bootstrap values based on 1000 bootstraps.
Insects 13 00405 g007
Table
Table 1. Population, date, and number (n) of wing images of S. calcitrans used for the landmark-based geometric morphometric analysis.
Table 1. Population, date, and number (n) of wing images of S. calcitrans used for the landmark-based geometric morphometric analysis.
PopulationCodeRegionDateBiotopeAltitudeCoordinates (Lat/Long)n
Mae Hong SonMHNorthernJulyBeef cattle farm located in rural area45219°31′45″ N, 98°04′48″ EFemale 50, Male 50
Nakhon RatchasimaNRNortheasternFebruaryBeef cattle and buffalo farm located in rural area49814°22′23″ N, 101°44′51″ EFemale 45, Male 45
Nakhon PathomNPCentralMayBeef cattle farm located in urban area1114°01′10″ N, 99°57′37″ EFemale 50, Male 50
KanchanaburiKBWesternMarchBeef cattle farm located in rural area17414°39′28″ N, 98°32′19″ EFemale 50, Male 50
SongkhlaSKSouthernFebruaryZoo located in urban area11207°08′26″ N, 100°36′20″ EFemale 50, Male 50
Table
Table 2. Mean CS of male and female S. calcitrans populations and statistically significant differences.
Table 2. Mean CS of male and female S. calcitrans populations and statistically significant differences.
PopulationnMean (mm)(Min–Max)VarianceSDSE
Female
MH504.43 a3.88−4.840.060.250.04
NR454.40 ac3.90−4.790.050.240.04
NP504.40 ad3.87−4.730.030.180.02
KB504.34 ab3.77−4.690.070.260.04
SK504.09 h3.37−4.550.080.280.04
Male
MH504.29 be3.71−4.630.050.230.03
NR454.23 efg3.55−4.610.050.220.03
NP504.30 bcdf3.75−4.590.040.200.03
KB504.14 gh3.57−4.590.050.210.03
SK503.97 i3.57−4.310.030.180.03
Statistically significant differences (p < 0.05) are indicated by different letters.
Table
Table 3. Mahalanobis distances (below diagonal) and p-values (above diagonal) among the wing shapes of female and male S. calcitrans populations.
Table 3. Mahalanobis distances (below diagonal) and p-values (above diagonal) among the wing shapes of female and male S. calcitrans populations.
PopulationMHNRNPKBSK
Female
MH-0.005<0.001<0.001<0.001
NR1.55-<0.001 <0.001<0.001
NP1.942.51-<0.001<0.001
KB3.222.963.69-<0.001
SK3.452.993.981.41-
Male
MH-<0.001<0.001<0.001<0.001
NR1.68-<0.001<0.001<0.001
NP2.522.26-<0.001<0.001
KB4.383.994.08-<0.001
SK4.694.354.171.61-
Table
Table 4. Percentage of correctly assigned individuals based on the cross-validated classification of the wing size and shape of female and male S. calcitrans populations.
Table 4. Percentage of correctly assigned individuals based on the cross-validated classification of the wing size and shape of female and male S. calcitrans populations.
PopulationSizeShape
% Correctly Assigned IndividualsNo. of Correctly Assigned Individuals/Total Numbers% Correctly Assigned IndividualsNo. of Correctly Assigned Individuals/Total Numbers
Female
MH5829/505226/50
NR00/4551.1123/45
NP21/507035/50
KB63/506633/50
SK168/505427/50
Male
MH126/505829/50
NR13.336/4553.3324/45
NP42/507236/50
KB189/505829/50
SK5226/508040/50
Total18.3790/49061.63302/490
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Chaiphongpachara, T.; Duvallet, G.; Changbunjong, T. Wing Phenotypic Variation among Stomoxys calcitrans (Diptera: Muscidae) Populations in Thailand. Insects 2022, 13, 405. https://doi.org/10.3390/insects13050405

AMA Style

Chaiphongpachara T, Duvallet G, Changbunjong T. Wing Phenotypic Variation among Stomoxys calcitrans (Diptera: Muscidae) Populations in Thailand. Insects. 2022; 13(5):405. https://doi.org/10.3390/insects13050405

Chicago/Turabian Style

Chaiphongpachara, Tanawat, Gerard Duvallet, and Tanasak Changbunjong. 2022. "Wing Phenotypic Variation among Stomoxys calcitrans (Diptera: Muscidae) Populations in Thailand" Insects 13, no. 5: 405. https://doi.org/10.3390/insects13050405

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop