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Evaluation of the wing cell contour to distinguish between Stomoxys bengalensis and Stomoxys sitiens (Diptera: Muscidae) using outline-based morphometrics.

ABSTRACT: The blood-sucking flies of the genus Stomoxys Geoffroy, 1762 (Diptera: Muscidae) are significant ectoparasites that can cause irritation and transmit pathogens to both animals and humans. Within the genus Stomoxys, two species, Stomoxys bengalensis and Stomoxys sitiens, have similar morphology and coexist in the same habitat. Accurate species identification of these flies is crucial for understanding disease vectors and implementing effective control measures. In this study, we assessed the effectiveness of outline-based geometric morphometrics (GM) by analyzing the wing cell contour of the first posterior cell (R₅) to distinguish between species and sexes of S. bengalensis and S. sitiens. Our results demonstrate that the outline-based GM method is highly effective in distinguishing between species and sexes of these flies based on contour shape, with accuracy scores ranging from 90.0% to 97.5%. Therefore, outline-based GM emerges as a promising alternative to landmark-based GM or as a supplementary tool in conjunction with traditional morphology-based methods for species identification.

SUBMITTER: Weluwanarak T

PROVIDER: S-EPMC11342272 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature

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Evaluation of the wing cell contour to distinguish between Stomoxys bengalensis and Stomoxys sitiens (Diptera: Muscidae) using outline-based morphometrics.

Weluwanarak Thekhawet T Chaiphongpachara Tanawat T Changbunjong Tanasak T

Current research in parasitology & vector-borne diseases 20240725

The blood-sucking flies of the genus Stomoxys Geoffroy, 1762 (Diptera: Muscidae) are significant ectoparasites that can cause irritation and transmit pathogens to both animals and humans. Within the genus Stomoxys, two species, Stomoxys bengalensis and Stomoxys sitiens, have similar morphology and coexist in the same habitat. Accurate species identification of these flies is crucial for understanding disease vectors and implementing effective control measures. In this ...[more]

PMID: 39185326

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Project description:Background and aimStomoxys and tabanid flies are of medical and veterinary importance because they play crucial roles in disease transmission as mechanical vectors of various hemopathogens. However, its role as a hemoprotozoan parasite vector in horse farms has not been studied. Therefore, we investigated the occurrence of hemoprotozoan parasites belonging to the genera Babesia, Theileria, and Trypanosoma in Stomoxys and tabanid flies using conventional polymerase chain reaction (PCR) and DNA sequencing.Materials and methodsAll samples (Stomoxys and tabanid flies) were collected using an Nzi trap for three consecutive days each month from November 2022 to March 2023. The flies were morphologically identified to the species level and separated according to sex. Individual (for tabanid flies) or pooled samples (consisting of three specimens of Stomoxys flies of the same species and sex collected from the same site) were used for DNA extraction. Conventional PCR was used to screen for hemoprotozoan parasite DNA, followed by Sanger sequencing to identify the species.ResultsIn total, 189 biting flies were collected, including four species of Stomoxys (Stomoxys bengalensis, Stomoxys calcitrans, Stomoxys indicus, and Stomoxys sitiens) and five species of tabanids (Atylotus cryptotaxis, Chrysops dispar, Tabanus megalops, Tabanus mesogaeus, and Tabanus rubidus). Stomoxys calcitrans was the most prevalent species, accounting for 58.7% (n = 111) of the collected flies. Ten (12.4%) of the 81 samples (individuals and pools) analyzed by PCR were positive for the 18S rRNA gene of the Theileria/Babesia species. Trypanosoma DNA was not detected in any sample. After performing Basic Local Alignment Search Tool searches and a phylogenetic analysis, only six samples (7.4%), including S. calcitrans (n = 2), S. sitiens (n = 2), T. megalops (n = 1), and A. cryptotaxis (n = 1), were found to be infected with Theileria sinensis. Furthermore, apicomplexan parasites, namely, Mattesia spp. and Colpodella spp., were found on S. indicus, the fungus Meira spp. was found on S. calcitrans, and the pathogenic green alga Helicosporidium spp. was found on A. cryptotaxis.ConclusionThis study is the first to report a variety of Stomoxys and tabanid flies collected from horse farms in Thailand, which were found to be infected with Theileria and Colpodella species that affect mammals, suggesting that Stomoxys and tabanid flies can be used to confirm the presence of hemoprotozoan parasites in the study area. Understanding the presence of hemoprotozoa in flies could help design vector control programs and manage various diseases in the study area.

Project description:BackgroundFast and reliable species identification of black flies is essential for research proposes and effective vector control. Besides traditional identification based on morphology, which is usually supplemented with molecular methods, geometric morphometrics (GM) has emerged as a promising tool for identification. Despite its potential, no specific GM techniques have been established for the identification of black fly species.MethodsAdult female black flies collected using human bait, as well as those reared from pupae, were used in this study. Here, landmark-based GM analysis of wings was assessed for the first time to identify human-biting black fly species in Thailand, comparing this approach with the standard morphological identification method and DNA barcoding based on the mitochondrial cytochrome c oxidase subunit I (COI) gene. To explore genetic relationships between species, maximum likelihood (ML) and neighbor-joining (NJ) phylogenetic trees were built. Additionally, three different methods of species delimitation, i.e., assemble species by automatic partitioning (ASAP), generalized mixed yule coalescent (GMYC), and single Poisson tree processes (PTP), were utilized to identify the morphologically defined species. The effectiveness of a COI barcode in identifying black fly species was further examined through the best match (BM) and best close match (BCM) methods.ResultsSeven black fly species, namely Simulium tenebrosum Takaoka, Srisuka & Saeung, 2018 (complex), S. doipuiense Takaoka & Choochote, 2005 (complex), S. nigrogilvum Summers, 1911, S. nodosum Puri, 1933, S. asakoae Takaoka & Davies, 1995, S. chamlongi Takaoka & Suzuki, 1984, and S. umphangense Takaoka, Srisuka & Saeung, 2017 were morphologically identified. Compared with the standard method, the GM analysis based on wing shape showed high success in separating species, achieving an overall accuracy rate of 88.54%. On the other hand, DNA barcoding surpassed wing GM for species identification with a correct identification rate of 98.57%. Species delimitation analyses confirmed the validity of most nominal species, with an exception for S. tenebrosum complex and S. doipuiense complex, being delimited as a single species. Moreover, the analyses unveiled hidden diversity within S. asakoae, indicating the possible existence of up to four putative species.ConclusionsThis study highlights the potential of wing GM as a promising and reliable complementary tool for species identification of human-biting black flies in Thailand.

Dataset Information

Evaluation of the wing cell contour to distinguish between Stomoxys bengalensis and Stomoxys sitiens (Diptera: Muscidae) using outline-based morphometrics.

Publications

Evaluation of the wing cell contour to distinguish between <i>Stomoxys bengalensis</i> and <i>Stomoxys sitiens</i> (Diptera: Muscidae) using outline-based morphometrics.

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