DNA barcoding and phylogenetic analysis of dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae) in wildlife and wildlife-livestock systems of Namibia

DNA barcoding and phylogenetic analysis of dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae) in wildlife and wildlife-livestock systems of Namibia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article DNA barcoding and phylogenetic analysis of dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae) in wildlife and wildlife-livestock systems of Namibia Mukendwa Hosticks Ndozi, Linnet Gohole, Isaac Mapaure, Emily Jepyegon Chemoiwa This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8627044/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 11 You are reading this latest preprint version Abstract Taxonomical keys have been used in the identification of dung beetle species due to its cost effectiveness and accessibilities. However, morphological identification of dung beetles remains a challenging task as a result of variable morphological differences among the species. The identification of dung beetles by the use of molecular markers helps in avoiding inaccuracy and appears promising to solve the problem of identifying species. The aim of this study was to determine the DNA barcoding and phylogenetic analysis of dung beetles in wildlife and wildlife-livestock ecosystems of Namibia. Sampling of dung beetles was done using baited pitfall traps and some species were handpicked from the abundant bovine dung pads. The collected dung beetles were preserved in DNA/RNA shield (Zymo Research), and kept in a freezer (-20 ℃) before the initial transportation. The genomic DNA was isolated using blood and tissue Quick-DNA Miniprep Plus Kit (Zymo Research). Two sets of universal primers based on the cytochrome c oxidase gene I (COI) barcodes selected for use. The sequence FASTA files were aligned using multiple alignments ClustalW software, and the phylogenetic relationships by Neighbor joining analysis was performed using the MEGA12 software. A total of 62 DNA barcodes of dung beetle species were accomplished from which 15 species from 27 sequenced samples were identified with the percentage of 95–100%, while 35 dung beetle sequences were not retrieved from the database. Indicating that most of the sequences found in wildlife and wildlife-livestock systems did not have reference sequences publicly entered into the database. The Neighbour-Joining phylogenetic tree showed 9 species of the genera Onthophagus , Milichus , and Caccobius of the tribe Onthophagini separated from each other and grouped together with the bootstrap value (> 95%). The clustering examination divided the 32 species into three main clades, while two species appeared separately from the three clades. This constituted the first report of DNA barcodes for Namibian dung beetles. The DNA barcoding and phylogenetic analysis application for molecular identification in dung beetles provides valuable and accurate information, which is useful in understanding their diversity. Scarabaeinae Molecular identification COI barcodes Figures Figure 1 Figure 2 Figure 3 Introduction Dung beetles are a group of insects that are regarded as coprophagous (mainly feeding on dung materials), and belong to the largest order of Coleoptera, family Scarabaeidae of the subfamily Scarabaeinae. The diversity of dung beetles is associated with the dung types excreted by mammals (Raine and Slade 2019 ), vegetation types, characteristics of the soil and seasonality of rainfall (Tshikae et al. 2008 ). This group of insects are globally distributed with 442,275 described species in 29,595 genera (Goczał et al. 2024 ), with more than 7,000 described species belonging to the subfamily of Scarabaeinae (Daniel and Davis 2024 ). About 540 species of dung beetles have been reported in the ecological zones of Southern African regions, especially in countries such as Botswana, Namibia and South Africa. Presently, 214 dung beetle species have been recorded in various ecosystems of Namibia, with 22 species have been identified as endemic species and majority of these endemic species are adapted to the Namib Desert (Davis et al. 2020). The presence of high species composition, diversity, abundance and assemblages of dung beetles in different ecosystem of Africa is primarily influenced by the presence of either abiotic or biotic factors (Daniel et al. 2022 ). Rainfall patterns, temperature fluctuations, elevation and soil types are some of the abiotic factors that alters the assemblage structures of dung beetles (Davis et al. 2020; Daniel et al. 2022 ). On the other hand, some of the biotic factors that changes the assemblages of dung beetles includes habitat types, vegetation cover, dung resources that are influenced by the diversity and abundance of mammalian species (Pryke et al. 2022 ; Bruda et al. 2024 ). Dung beetles are sensitive to environmental changes that are inclined as a result of habitat modifications and anthropogenic actions, one of the reasons why these insects are primarily considered as bioindicators (Davis et al. 2001 ; Nichols et al. 2008 ). Environmental disturbances have been causing a serious replacement of native flora by large areas of agriculture and pasture (Nichols et al. 2007 ). Loss in biodiversity as a result in the reduction of natural areas is the main leading factor to loss of genetic variability of the remaining species. Dung beetles are specialist in utilising dung pads for feeding, breeding and nesting (Heddle et al. 2021 ; Thotagamuwa et al. 2023 ). Several ecological functions and ecosystem services are complimented by dung beetles when utilising dung pads. The ecological processes that are driven by the dung beetles during the relocation and consumption of dung materials include the improvement of soil aeration, nutrient cycling, secondary seed burial, parasite suppression, bioturbation and fly control (Andresen 2002 ; Nichols et al. 2008 ; Arellano et al. 2023 ). Morphological identification of dung beetles remains a challenging task as a result of variable morphological differences among the species (Murphy 2020). Some of the morphological differences appears in different life stages of the species, for instance, it is somehow challenging to identify the larval stage of some insects. However, the identification keys on dung beetles using morphological characteristics are more effective on a particular life stage such as the adult stage (Bandral et al. 2022 ; Jagdale et al. 2023 ). Therefore, challenges related to the effective use of the taxonomic keys can be resolved with the use of DNA barcoding. The identification of dung beetles by the use of molecular markers is useful in eliminating inaccuracy and appears promising in solving the problem of misidentification of species (Johnston et al. 2025 ). DNA barcoding has different techniques that have been adopted for the identification and characterization of scarab beetles, which include the use of mitochondrial DNA (mtDNA)-encoded cytochrome oxidase I genes (COI) (Mlambo et al. 2015 ; Baena-Bejarano et al. 2023 ; Watanabe et al. 2024 ). Villalba et al. ( 2002 ) reported the first use of molecular identification in the subfamily of Scarabaeinae, whereas mitochondrial cytochrome oxidase I and II (COI, COII) were considered for PCR and DNA sequencing. The African continent is characterised by the best Scarabaeinae dung beetle diversity; a total of 2,000 species have been documented (Scholtz et al. 2009 ), 670 species inhabit Southern Africa, and 214 species have been reported in Namibia (Davis et al. 2020). Though DNA barcoding and phylogenetic analysis have been performed in Southern Africa (where Namibia is located), research using DNA to identify dung beetles has not been fully exploited in Namibia. Considering that the overall distribution and knowledge of DNA barcoding and phylogenetic analysis in the Namibian dung beetle community on a global scale are poor, the molecular study of the Scarabaeinae is still in need of treatment and recommendation of gene sequences to bridge this gap. The use of molecular data needs attention in order to address the problem of misidentification of each of these insects, focusing on species-level identification and the exploitation of molecular information to make a significant difference. Dung beetles are considered a cosmopolitan insect family with vital service roles in ecosystems (Davis and Scholtz 2001), and phylogenetic determination will help establish their phylogenetic positions and classify them into clades based on their tribal association. Since dung beetles of Namibia are under-investigated in molecular sequencing areas, this creates a significant gap in the global study of biogeography, distribution, and diversity of dung beetles. For example, Namibia has been able to document at most nine dung beetle tribes from morphological analysis (Davis et al. 2020; Ndozi et al. 2025 a). In a similar manner, the relationship of evolution, as reflected in the Bayesian phylogram, showed that there are nine African Scarabaeinae tribes, which include Ateuchini, Coprini, Canthonini, Gymnopleurini, Oniticellini, Onitini, Onthophagini, Scarabaeini, and Sisyphini. This phylogenetic relationship is critical in the study of dung beetles and also in understanding genetic flow, identification, and population organization. Therefore, the study determined the DNA barcoding and phylogenetic analysis of the dung beetles in wildlife and wildlife-livestock systems of Namibia. Thus, phylogeographic studies and biodiversity assessments are therefore supported by the correct species identification, phylogenetic position of the species, and the availability of the DNA barcodes. Such kind of information has not documented with reference to Namibian dung beetles. Methods and Materials Study area The study was conducted at Nkasa Rupara National Park (23.607676 E; 18.364642 S), which is a wildlife system, and Dzoti Conservancy (23.781766 E; 18.252163 S), which is the wildlife-livestock system (Fig. 1 ). Both study sites are situated in the north-eastern part of Namibia. The selection of the two systems was mainly based on the similarities of the vegetation, soil type, and rainfall seasonality. The national park is characterised by the riparian woodland and grassland that mostly dominant in floodplain areas, and the park covers a distance of 900 km 2 (MEFT, 2020). While Dzoti Conservancy is a wildlife-livestock system covering an area of 287 km2 and is home to an estimated human population of over 2,000 (Denker 2020 ). Woodland vegetation is mainly dominated by the broadleaved trees including tree species such as Combretum imberbe (Wawra 1860), Kigelia africana (Lamarck 1849), Senegalia nigrescens (Oliver et al. 1871), Vachellia sieberiana (de Candolle 1825), Albizia versicolour (Oliver 1871), Piliostigma thonningii (Schumach 1947), and Terminalia sericea (Burchell 1828). The floodplain areas are dominated by the grass species of Hyparrhenia rufa (Stapf 1919), other non-woody species adapted in floodplains includes reeds, edges and wild date palms. The vegetation types along the riverbanks and channels are dominated by broadleaved trees such as Diospyros mespiliformis (Hochstetter 1844) and Garcinia livingstonei (Anderson1866). The mammalian species that are common in the national park includes the cape buffalo, elephant, wildebeest, hippopotamus and several others species. The cambisol soil types dominates national park (FAO and ITPS 2015). Rain begins to fall in late November and lasts until mid-March, with an annual rainfall ranging from 400–800 mm and temperatures ranging from 24–28°C. In winter, minimum temperature ranges between 2°C and 8°C, while in summer, maximum temperature can range between 36°C and 40°C (Ndozi et al. 2025 a; 2025 b). Sampling of dung beetles Dung beetles were sampled either by hand picking from the abundant bovine dung pads, whereas some of the dung beetles were collected using pitfall traps. For small (less than 2 mm) dung beetle species, the entire insect was placed in a 2 ml specimen vial with DNA Shield solution, with a maximum of 4 individuals placed in each vial. Dung beetles that had a lot of dung materials on their exoskeletons were either excluded or washed with deionized water to remove the excrements. In large specimens (2 mm – 5 mm), the middle femurs were removed, then sliced into small pieces and placed in a labelled specimen vial. Other parts such as the abdomen, head and thorax were excluded in larger species as they may contain dung materials that might interfere with the process of DNA extraction. Preservation of specimens All the dung beetle species that were collected from both systems were preserved in DNA/RNA shield (Zymo Research). After placing the samples in DNA/RNA shield, samples were kept in a freezer (-20 ℃) before the initial transportation. All the collected samples were transported to the Fish Genomic and Genetic Research Laboratory (FGGRL). At the Fish Genomic and Genetic Research Laboratory (FGGRL), University of Eldoret, samples were transferred to the freezer at -20 ℃ temperature. The DNA extraction and PCR amplification was conducted at the FGGRL. Pretreatment of samples and DNA extraction DNA from each species of dung beetle was extracted separately using the blood and tissue Quick-DNA Miniprep Plus Kit (Zymo Research) according to the protocols of the manufacturer in exception of few modifications as described below. Prior to the extraction, samples (30 mg) were placed in 1.5 ml eppendorf tubes followed by the addition of warm double-distilled water (30°C), and vortexing was done for 15 seconds. After three times of washing, samples were crushed in the eppendorf tubes using Eppendorf micropestles. DNA quantification The extracted DNA was quantified using the Nanodrop spectrophotometer (Thermo Fisher Scientific) to verify the concentration (ng/µl) of each DNA sample. The determination of purity of DNA was performed by taking the ratio of the sample absorbance at 260 and 230 nm (260/230) and also 260 and 280 nm (260/280). Polymerase Chain Reaction (PCR) amplification The amplifications were carried out with mtDNA using two sets of universal primers based on the cytochrome c oxidase gene I (COI) barcodes (Table 1 ). The PCR reactions were carried out in clear 0.2 ml microtubes with caps. The total PCR reaction volume of 25 µl was used: 6.5 µl was nucleic acid-free water; 12.5 µl was master mix; 0.5 µl was the forward primer; 0.5 µl was the reverse primer; and 5 µl consisted of template DNA (Asha and Sinu 2021 ). mtDNA cytochrome c oxidase gene I (COI) barcoding universal primers were added: COI forward primer (C1-J-2183): 5’CAACATTTATTTTGATTTTTTGG-3’ and COI reverse primer (TL2-N-3014): 5’TCCAATGCACTAATCTGCCATATTA-3’ (Simon et al. 1994) were used. PCR amplification was done using the following conditions: The initial denaturation temperature was set at 94°C for 90 seconds, followed by denaturation at 94°C for 22 seconds, followed by 35 cycles, annealing at 50°C for 30 seconds, extension at 72°C for 90 seconds, and the final extension was at 72°C for 60 seconds (Mlambo et al. 2011). For the second primers, the PCR reaction was performed following the established protocol similar to the first primers using COI forward primer (LCO1490): 5’GGTCAACAAATCATAAAGATATTGG-3’ and COI reverse primer (HCO2198): 5’TAAACTTCAGGGTGACCAAAAAATCA-3’ (Folmer et al. 1994). PCR amplification was done using the following conditions: The initial denaturation temperature was set at 95°C for 120 seconds, followed by denaturation at 95°C for 30 seconds, followed by 34 cycles, annealing at 50°C for 60 seconds, extension at 72°C for 2 minutes, and the final extension was at 72°C for 7 minutes (Gavarāne et al. 2011 ). The amplicons were confirmed by the gel electrophoresis using 1% agarose (Fig. 2 ). The amplified products were then sent to Inqaba Biotec East Africa (Ltd) in Nairobi, where sequencing was done. A total of 62 sequences of 740 base pairs (bp) representing 33 species of 23 genera ( Allogymnopleurus , Caccobius , Catharsius , Chalconotus , Copris , Digitonthophagus , Euonthophagus , Gymnopleurus , Kheper , Kurtops , Latodrepanus , Metacatharsius , Milichus , Oniticellus , Onitis , Onthophagus , Pachylomera , Phalops , Scarabaeus , Sisyphus , and Tiniocellus ) were taken along with 1 species of Epirinus aeneus (GQ290002) using the COI gene fragment for phylogenetic analysis belonging to the subfamily of Scarabaeinae. The species E. aeneus of tribe Deltochilini of the same subfamily was used as an outroot for using the COI gene fragment for phylogenetic analysis. The species E. aeneus of the subfamily Scarabaeinae is regarded to be an endemic species of southern Africa. The construction of the phylogenetic tree was based on COI gene sequences using 1,000 bootstrapped Neighbour-Joining with the Kimura-2 parameters. Data analysis The sequence FASTA files were aligned using multiple alignments ClustalW software and edited using the BioEdit Sequence Alignment Editors v 7.2; this was performed to eliminate components that dispersed and bases that were unclear. After aligning and trimming the sequences, their length was 786bp. The similarities of the sequences were aligned using the BLAST algorithm, and a database of 62 sequences was created. The collection details of all the sequences that were uploaded to the Bold Systems and accession numbers recorded. The MEGA software version 12.0 (Kumar et al., 2024 ) was used in performing the phylogenetic relationships by Neighbor Joining (NJ) analysis whereby Kimura’s two-parameter substitution model was used, with a consideration of 1,000 bootstrap replicates (Tamura et al. 2021 ). FigTree version 1.4.4 was used to visualize the phylogenetic tree. Results DNA barcoding All the sequences for wildlife and wildlife-livestock systems samples were compared by performing BLAST analysis with those sequences in the NCBI GenBank. The present study compared the barcode of 61 mtDNA amplicons of the genera Allogymnopleurus , Caccobius , Catharsius , Chalconotus , Euoniticellus , Euonthophagus , Gymnopleurus , Heliocopris , Kheper , Latodrepanus , Metacatharsius , Milichus , Onitis , Onthophagus , Pachylomera , Phalops , Scarabaeus , Sisyphus , and Tiniocellus dung beetles submitted to Bold Systems. The respective species were identified to genus and species level. From a total of 27 DNA sequences which matched in the NCBI GenBank, 15 species were identified with the percentage of 95–100% identity distributed in to 14 genera. In the 27 sequences identified, twelve species appeared common from the two study ecosystems in Namibia (Table 1 ). Approximately 34 dung beetle species identified morphologically as earlier reported by Ndozi et al. (2024) were not retrieved and identified from the NCBI database (Table 2 ). The unretrieved were 24 species belonging to 11 genera showing a high population of Namibian dung beetles as being under reported. Table 1 Scarabaeinae taxa studied in wildlife and wildlife-livestock ecosystems and their Bold Systems accession numbers, percent identity and COI primers. Lab ID Species name BOLD systems Accession Numbers Primer set used Percent identity Matched Accession Numbers in NCBI C1 Allogymnopleurus thalassinus (Klug,1855) NAMC020 TL2N3014/C1J2183 99 AY131898.1 C15 Latodrepanus laticollis (Fahraeus, 1857) NAMC023 TL2N3014/C1J2183 99 EF188137.1 C16 Metacatharsius opacus (Waterhouse, 1891) NAMC004 TL2N3014/C1J2183 98 AY131865.1 C17 Milichus apicalis (Fahraeus, 1857) NAMC024 TL2N3014/C1J2183 97 AY131921 C18 Onitis alexis (Klug, 1835) NAMC017 HCO2198/LCO1490 95 HM375977.1 C24 Onthophagus variegatus (Erichson, 1843) NAMC019 TL2N3014/C1J2183 99 EF188218 C26 Pachylomera femoralis (Kirby, 1828) NAMC007 TL2N3014/C1J2183 100 JN804658.1 C29 Sisyphus goryi Harold, 1859 NAMC005 TL2N3014/C1J2183 99 MH129898.1 C3 Chalconotus convexus (Boheman, 1857) NAMC025 TL2N3014/C1J2183 98 AY131809.1 C31 Tiniocellus spinipes (Roth, 1851) NAMC011 TL2N3014/C1J2183 97 AY131912.1 C32 Scarabaeus zambesianus Péringuey, 1901 NAMC010 TL2N3014/C1J2183 99 AF499770.1 C7 Digitonthophagus gazella (Fabricius, 1787) NAMC016 TL2N3014/C1J2183 99 AY131918.1 C9 Euonthophagus carbonarius (Klug,1855) NAMC001 TL2N3014/C1J2183 99 AY131919 N1 Allogymnopleurus thalassinus NAMC018 TL2N3014/C1J2183 100 AY131898.1 N11 Euonthophagus carbonarius (Klug, 1855) NAMC012 TL2N3014/C1J2183 99 AY131919 N17 Kurtops signatus (Fahraeus, 1857) NAMC026 TL2N3014/C1J2183 99 EF188216 N18 Latodrepanus laticollis (Fahraeus, 1857) NAMC022 TL2N3014/C1J2183 99 EF188137.1 N19 Metacatharsius opacus (Waterhouse, 1891) NAMC002 TL2N3014/C1J2183 96 AY131865.1 N20 Milichus apicalis (Fahraeus, 1857) NAMC013 TL2N3014/C1J2183 99 AY131921.1 N22 Onitis alexis (Klug, 1835) NAMC027 HCO2198/LCO1490 96 KF801856.1 N27 Onthophagus fimetarius (Roth, 1851) NAMC014 TL2N3014/C1J2183 100 AY131925 N32 Onthophagus variegatus (Erichson, 1843) NAMC021 TL2N3014/C1J2183 99 EF188217 N35 Pachylomera femoralis (Kirby, 1828) NAMC008 TL2N3014/C1J2183 99 JN804658.1 N39 Sisyphus goryi Harold, 1859 NAMC028 TL2N3014/C1J2183 98 MH129898.1 N41 Tiniocellus spinipes (Roth, 1851) NAMC015 TL2N3014/C1J2183 98 AY131912.1 N43 Scarabaeus zambesianus Péringuey, 1901 NAMC009 TL2N3014/C1J2183 99 AF499770.1 N9 Digitonthophagus gazella (Fabricius, 1787) NAMC003 TL2N3014/C1J2183 100 AY131918 Table 2 Unretrieved dung beetle sequences in gene bank database and BOLD systems. La ID Species name BOLD systems Accession Numbers Primer set used C11 Kheper lamarcki (Macleay, 1821) NAMC029 TL2N3014/C1J2183 C12 Kheper nigroaeneus (Boheman, 1857) NAMC030 TL2N3014/C1J2183 C19 Onitis deceptor (Péringuey, 1901) NAMC031 TL2N3014/C1J2183 C2 Catharsius aegeus (Génier, 2017) NAMC032 TL2N3014/C1J2183 C21 Onthophagus lamelliger (Gerstaecker, 1871) NAMC033 TL2N3014/C1J2183 C22 Onthophagus quadrinotatus (d'Orbigny, 1905) NAMC034 TL2N3014/C1J2183 C25 Onthophagus vinctus (Erichson,1843) NAMC035 TL2N3014/C1J2183 C27 Phalops flavocinctus (Klug, 1855) NAMC036 TL2N3014/C1J2183 C28 Kheper prodigiosus (Erichson, 1843) NAMC037 TL2N3014/C1J2183 C33 Heliocopris japetus (Klug, 1855) NAMC038 TL2N3014/C1J2183 C34 Catharsius tricornutus (DeGeer, 1778) NAMC039 TL2N3014/C1J2183 C4 Copris amyntor (Klug, 1855) NAMC006 TL2N3014/C1J2183 C5 Copris elphenor (Klug, 1855) NAMC040 TL2N3014/C1J2183 C6 Copris puncticollis (Boheman, 1857) NAMC041 TL2N3014/C1J2183 C10 Gymnopleurus pumilus (Reiche, 1850) NAMC061 HCO2198/ LCO1490 N10 Euoniticellus intermedius (Reiche, 1849) NAMC059 HCO2198/ LCO1490 N12 Gymnopleurus pumilus (Reiche,1850) NAMC042 HCO2198/LCO1490 N14 Kheper lamarcki (Macleay, 1821) NAMC060 TL2N3014/C1J2183 N15 Kheper nigroaeneus (Boheman, 1857) NAMC043 TL2N3014/C1J2183 N2 Caccobius nigritulus (Klug, 1855) NAMC044 TL2N3014/C1J2183 N21 Oniticellus formosus (Chevrolat, 1830) NAMC045 TL2N3014/C1J2183 N23 Onitis deceptor (Péringuey, 1901) NAMC046 TL2N3014/C1J2183 N24 Onthophagus aeruginosus (Roth, 1851) NAMC047 TL2N3014/C1J2183 N28 Onthophagus flavolimbatus (Klug, 1855) NAMC048 TL2N3014/C1J2183 N29 Onthophagus lamelliger (Gerstaecker, 1871) NAMC049 TL2N3014/C1J2183 N3 Catharsius aegeus (Génier, 2017) NAMC050 TL2N3014/C1J2183 N30 Onthophagus quadrinotatus (d'Orbigny, 1905) NAMC051 TL2N3014/C1J2183 N31 Onthophagus suffusus (Klug, 1855) NAMC052 TL2N3014/C1J2183 N34 Onthophagus vinctus (Erichson, 1843) NAMC053 TL2N3014/C1J2183 N36 Phalops boschas (Klug, 1855) NAMC054 TL2N3014/C1J2183 N37 Phalops flavocinctus (Klug, 1855) NAMC055 TL2N3014/C1J2183 N38 Kheper prodigiosus (Erichson, 1843) NAMC056 TL2N3014/C1J2183 N7 Copris elphenor (Klug, 1855) NAMC057 TL2N3014/C1J2183 N8 Copris puncticollis (Boheman, 1857) NAMC058 TL2N3014/C1J2183 Phylogenetic analysis of dung beetles The Neighbour-Joining phylogenetic tree (Fig. 3 ) showed that 9 species of the genera Onthophagus , Milichus , and Caccobius of the tribe Onthophagini separated from each other and grouped together with the bootstrap value (> 95%). Digitonthophagus gazella , Euonthophagus carbonarius , Kurtops signatus and Onthophagus flavolimbatus of the similar tribe Onthophagini also separated and grouped together with the bootstrap value (> 95%). The clustering examination divided the 32 species into three main clades, while two species (NP37 Phalops flavocinctus and NP21 Oniticellus formosus) appeared separately from the three clades. The first clade consisted mainly of species from the genera Onthophagus , Milichus , Caccobius , Latodrepanus , Onitis , Sisyphus , Tiniocellus , Kurtops , Euonthophagus , and Digitonthophagus . The second clade was composed of species of the genera Allogymnopleurus , Gymnopleurus , Copris , Metacatharsius , Phalops , Pachylomera , Kheper , and Scarabaeus . The third clade included two species: Catharsius aegeus and Catharsius tricornutus . Discussion The present study provided the DNA barcodes of 62 species of the genera Allogymnopleurus , Caccobius , Catharsius , Chalconotus , Euoniticellus , Euonthophagus , Gymnopleurus , Heliocopris , Kheper , Latodrepanus , Metacatharsius , Milichus , Onitis , Onthophagus , Pachylomera , Phalops , Scarabaeus , Sisyphus , and Tiniocellus dung beetles. Dung beetles from different ecological zones of Namibia are poorly documented in the GenBank data base such as the BOLD systems and NCBI, hence some of the sequences (a total of 35 dung beetle species) were not identified to their respective species level However, the woodland savanna and mixed grassland ecosystems spanning the entire north-eastern part of Namibia are less explored for invertebrates including arthropods, despite having favorable microclimates that might support the species diversity and richness of different insect communities. The distribution and phylogenetic origin of dung beetles in both wildlife and wildlife-livestock systems appear to be unclear, contributing to the poor representation of dung beetles of Namibia in the global DNA barcoding, phylogenetic and biogeographical investigations and therefore the GenBank data base. Around 85% of the BLAST sequences compared were similar to dung beetles from the Sub-Saharan biogeographical region. Most of the sequences found in wildlife and wildlife-livestock systems did not have reference sequences publicly entered into the database. The current study is the first report by researchers of DNA barcodes for Namibian dung beetles. Little has been done regarding molecular identification, especially in the tribe of Onthophagini. Onthophagus flavolimbatus Klug, 1855, for example, one of the species from this tribe first appeared in the wildlife ecosystem of the northeastern part of Namibia. However, South Africa (Tocco et al. 2021 ) and Mozambique (Daniel and Génier 2019 ) reported this species to be present in parts of these countries, even though there was no DNA sequence for it in this database. The tribe Onthophagini, to which O. flavolimbatus belongs, as well as other tribes like Sisyphini, Oniticellini, and Coprini, are considered cosmopolitan (Davis and Scholtz 2001). Apart from O. flavolimbatus , the study further provided insights on the first mitochondrial cytochrome c oxidase subunit 1 (COI) gene sequences of the species, including Copris elphenor Klug 1855, Heliocopris japetus Klug, 1855, Kheper lamarcki Macleay 1821, Oniticellus formosus Chevrolat 1830, Onthophagus aeruginosus Roth 1851, Onthophagus lamelliger Gerstaecker 1871, Onthophagus vinctus Erichson 1843, and Phalops flavocinctus Klug 1855. The DNA barcodes of these species have not yet been uploaded in the BOLD systems and GenBank of the NCBI database, making it difficult to identify these species using the molecular marker. There are three genera that represents tribe Gymnopleurini in Africa (Davis et al. 2020), however, the present study only captured two of the genera belonging to this tribe, namely Allogymnopleurus and Gymnopleurus . The findings of this study did not report on genus Garreta , which is the third genus in the tribe Gymnopleurini. Phylogenetic analysis also revealed that Gymnopleurini is a sister group to Scarabaeini, which indicates that this tribe is more closely related to the genus Pachylomera in that sister group. Based on phylogenetic analyses carried out elsewhere, the tribe has also been placed in a similar clade to the tribe Scarabaeini, while phylogenetic studies from other sources also show that the tribe Gymnopleurini is in a clade with Scarabaeini, hence demonstrating a strong sister relationship (Villalba et al. 2002 ; Monaghan et al. 2007 ; Mlambo et al. 2011). Gymnopleurini pumilus Reiche 1850, was not found in the database (unlike the species Allogymnopleurus thalassinus Klug 1855), indicating insufficient records on the phylogenetic work of the tribe Gymnopleurini. A relationship was revealed in the dung beetle species of Metacatharsius opacus Waterhouse 1891, within the tribe of Coprini and species from the tribe Sisyphini ( Sisyphus goryi Harold 1859). Philips et al. ( 2004 ) and Mlambo et al. (2011) displayed a similar phylogenetic relationship in species of the genus Metacatharsius . In addition, the species from the tribe Coprini are regarded as tunnelers, while those species in the tribe of Sisyphini are representing all the rollers. A monophyletic lineage was noticed in the genus of Onitis , showing a close relationship with the dung beetle species from the genera Tiniocellus and Latodrepanus . Additionally, tribe Oniticellini and Onthophagini, both showed a monophyletic lineage to some of the species within the tribe of Onitini. Mlambo et al. (2011) explained that the genera of Cheironitis and Onitis , all from the tribe Onitini, indicated a sister relationship with majority of the species from the tribe Onthophagini. According to Villalba et al. ( 2002 ) discovered an exciting understanding of the sisterhood relationship between the tribes Onitini and Onthophagini. Several molecular phylogeny studies have revealed the sister relationship of tribe Onitini are closely related to tribes Onthophagini and Oniticellini (Villalba et al. 2002 ; Monaghan et al. 2007 ). In addition, Philips et al. ( 2004 ) indicated that Onthophagini ( Digitonthophagus ), Onitini ( Babas , Heteronitis and Onitis ), and Oniticellini ( Tiniocellus ) were in one clade, showing the sister relationship. Dung beetles that belong to the tribe Onthophagini showed paraphyletic grouping; for instance, the species Digitonthophagus gazella Fabricius 1787, Euonthophagus carbonarius Klug 1855, Kurtops signatus Fahraeus 1857, and Onthophagus flavolimbatus Klug 1855, formed a clade that is sister to the rest of the genera (Villalba et al. 2002 ; Breeschoten et al. 2016 ; Philips et al. 2016; Zhang et al. 2025 ). In addition, the paraphyletic grouping within the tribe Onthophagini is due to presence of tribe Oniticellini. The results of this study revealed inconsistency of the genus Onthophagus within the tribe Onthophagini in their evolutionary clade. Onthophagus is presently categorized as highly polyphyletic among all the genus within the tribe of Onthophagini. Species such as Caccobius nigritulus , and Milichus apicalis were nested within the genus Onthophagus , despite the two genera been scarcely defined. The analysis confirmed the polyphyletic status of tribe Onthophagini within the main clade, which might have been influenced by the rearrangement among the terminal taxa, without changing their relationships and main lineage. Tribe Coprini was represented mainly by three genera ( Catharsius , Copris , and Metacatharsius ); species from the two genera, Copris and Metacatharsius , appeared to be a sister taxon, sharing suitable synapomorphies that might accurately define the tribe Coprini. In contrast, the species that belongs to Catharsius showed a homoplasy (Santis 2024 ) that they do not share a common ancestral root, despite being classified within the same tribe of Coprini. The species Pachylomera femoralis Kirby 1828, the only species representing the genus Pachylomera from the tribe Scarabaeini demonstrated a close relationship to Scarabaeus zambesianus Péringuey 1901. Both Pachylomera femoralis and Scarabaeus zambesianus are from the same tribe. Taxonomists have been proposing that the species Pachylomera femoralis should be a subgenus to Scarabaeus (Harrison et al. 2003; Forgie et al. 2006 ), while other studies have supported the current genus of Pachylomera (Scholtz et al. 2011). This study fully supports the classification of Pachylomera as a genus. Furthermore, Kheper lamarcki Macleay 1821, Kheper nigroaeneus Boheman 1857, and Kheper prodigiosus Erichson 1843, the only three species representing the genus Kheper exhibited a sister relationship with Scarabaeus zambesianus in this study confirming its classification as a genus. The results obtained by Mlambo et al. ( 2015 ) showed a close grouping of the species from the genera Kheper and scarabaeus , supporting their classification as separate genera. Hence, all the dung beetles belonging to the three genera ( Kheper , Pachylomera , and Scarabaeus ) of the tribe Scarabaeini are regarded as rollers. Rollers are group of dung beetles with the nesting behaviours of forming a brood ball that is pushed a distance from the dung pad and buried into the soil. The use of the COI gene can effectively disparate the numerous scarabs of Namibia, including the scarabs of southern Africa and other species that are found elsewhere in the world. Therefore, the application of molecular identification in dung beetles is valuable for those species that are challenging to be identified using morphologic characters and assists in identifying the unidentified dung beetle species. Conclusion DNA barcoding and phylogenetic analysis of dung beetles in wildlife and wildlife-livestock systems provided accurate identification at the species level, creating opportunities for future investigations. The use of the COI gene for species identification, which was adopted for this study, remains the technique for identifying species-level characteristics. Furthermore, the study complements mitochondrial COI sequences of several species of dung beetles to the database, thereby making these mitochondrial barcodes available for further studies in the same field. The COI gene is recommended as a more effective marker for DNA barcoding and phylogenetic analysis in dung beetle species; however, conducting a comprehensive investigation of population genetics using other genes is essential to confirm the species that have appeared and been identified morphologically in Namibia for the first time. Declarations The authors (Mukendwa Hosticks Ndozi, Linnet Gohole, Isaac Mapaure, Emily Jepyegon Chemoiwa) state and declare that there are no competing interests and this work has not been published elsewhere. Acknowledgments We thank the University of Eldoret through the Fishery Genomics and Genetic Laboratory for allowing us to conduct the DNA extraction and PCR amplification. Many thanks goes to Inqaba East Africa, who assisted with sequencing and other helpful support. Author contribution This manuscript was prepared through a collective effort between the listed co-authors; all the listed authors contributed to the research design, collection of data, data analysis, literature review, and revision of the manuscript. All the listed authors played a significant role in completing this manuscript. Funding information This research was funded by the University of Namibia under the office of the Pro-Vice Chancellor- Research, Innovation and Development via a scholarship awarded to the lead author (Mukendwa Hosticks Ndozi). References Andresen E (2002) Dung beetles in a Central Amazonian rainforest and their ecological role as secondary seed dispersers. 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Mol Biol Evol 41(12):msae263 Ministry of Environment, Forestry and Tourism (2020) Revised National Strategy on Wildlife Protection and Law Enforcement (2021–2025) Mlambo S, Sole CL, Scholtz CH (2015) A molecular phylogeny of the African Scarabaeinae (Coleoptera: Scarabaeidae). J Arthropod Syst Phylogeny 73(2):303–321 Monaghan MT, Inward DG, Hunt TH, Vogler AP (2007) A molecular phylogenetic analysis of the Scarabaeinae (dung beetles). Mol Phylogenetics Evol 45:674–692 Ndozi MH, Gohole L, Mapaure I (2025) Comparison of species diversity, richness, and abundance of dung beetles between wildlife and wildlife-livestock systems in north-eastern Namibia. Int J Trop Insect Sci, 1–14 Ndozi MH, Gohole L, Mapaure I (2025) Seasonal Variations in Dung Beetle (Scarabaeidae: Scarabaeinae) Assemblage in Wildlife and Wildlife–Livestock Systems in North-Eastern Namibia. Afr J Ecol, 63(6), e70083 Nichols E, Larsen T, Spector S, Davis ALV, Escobar F, Favila M, Network TSR (2007) Global dung beetle response to tropical forest modification and fragmentation: a quantitative literature review and meta-analysis. Biol Conserv 137(1):1–19 Nichols E, Spector S, Louzada JNC, Larsen TH, Amezquita S, Favila ME (2008) Ecological functions and ecosystem services provided by Scarabaeinae dung beetles. J Biol Conserv 461–1474 Philips KT, Pretoris E, Scholtz CH (2004) A phylogenetic analysis of dung beetles (Scarabaeinae: Scarabaeidae): unrolling an evolutionary history. – Invertebrate Systematics 18:53–88 Philips TK (2016) Phylogeny of the Oniticellini and Onthophagini dung beetles (Scarabaeidae, Scarabaeinae) from morphological evidence. ZooKeys, p 9. 579 Pryke JS, Roets F, Samways MJ (2022) Large African herbivore diversity is essential in transformed landscapes for conserving dung beetle diversity. J Appl Ecol 59:1372–1382 Raine EH, Slade EM (2019) Dung beetle–mammal associations: Methods, research trends and future directions. Proceedings of the Royal Society B: Biological Sciences, 286(1897), 20182002 Santis MD (2024) Homoplasy as an evolutionary process: an optimistic view on the recurrence of similarity in evolution. Biol Theory 19(4):267–278 Scholtz CH, Davis ALV, Kryger U (2009) Evolutionary Biology and Conservation of Dung Beetles. Pensoft, Pretoria, South Africa Tamura K, Stecher G, Kumar S (2021) MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol 38:3022–3027 Thotagamuwa A, Noriega JA, Webb S, Weston P, Doube BM, Caron V, Gurr GM (2023) Rearing dung beetles (Coleoptera: Scarabaeidae): identifying knowledge gaps and future challenges. Entomol Generalis 43(4):751–769 Tocco C, Dacke M, Byrne M (2021) The finely defined shift work schedule of dung beetles and their eye morphology. Ecol Evol 11(22):15947–15960 Tshikae BP, Davis ALV, Scholtz CH (2008) Trophic Associations of a Dung Beetle Assemblage (Scarabaeidae: Scarabaeinae) in a Woodland Savanna of Botswana. Environ Entomol 37(2):431–441 Villalba S, Lobo JM, Martín-Piera F, Zardoya R (2002) Phylogenetic relationships of Iberian dung beetles (Coleoptera: Scarabaeinae): insights on the evolution of nesting behavior. J Mol Evol 55(1):116–126 Watanabe S, Masamura N, Satoh SY, Hirao T (2024) Evaluating the Effectiveness of DNA Barcoding for Insect Identification: A Comprehensive Review. Entomol Lett 4(2–2024):34–41 Zhang H, He Q, Zhao Z, Zhang B, Zhou J, Wang C, Zhao M (2025) Mitochondrial Genome Comparison and Phylogenetic Analysis of Four Species of Dung Beetles (Coleoptera: Scarabaeidae: Scarabaeinae). Ecol Evol, 15(8), e71906 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 16 May, 2026 Reviews received at journal 15 May, 2026 Reviews received at journal 07 May, 2026 Reviewers agreed at journal 24 Apr, 2026 Reviewers agreed at journal 22 Apr, 2026 Reviews received at journal 08 Apr, 2026 Reviewers agreed at journal 06 Apr, 2026 Reviewers invited by journal 05 Apr, 2026 Editor assigned by journal 21 Jan, 2026 Submission checks completed at journal 21 Jan, 2026 First submitted to journal 17 Jan, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8627044","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":619910229,"identity":"27c06be4-b8b5-41c9-b3d5-3b826f74dce9","order_by":0,"name":"Mukendwa Hosticks Ndozi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6klEQVRIie3PMQuCQBTA8TscGvsqOkmTH6TlHUIt3u5gcVMufoCGqK9gi9BmHJzLQWvQYkuttTn2LoggMGsLur/wfMj94CTEZvvBXDMAHzMJxPh2HNFFqHgSbQj9gJAHoTPzrYP4vfRc18ko8HuhvByXk2E/RdLERSsZZNoToCK2yU7gsqLic0kFzfSh/WL7iAomYsDFBVYoLpA4dPaOjI+GBIaUbKH4qpuAhySi+T4yS8LzTqK1Nwc1Yrk+hQRUyddItm//pUrra5OEgVuFEpcpX+7ktm7idvKavM/y4/PY9JvDNpvN9ifdALDZZm944L0/AAAAAElFTkSuQmCC","orcid":"","institution":"University of Eldoret","correspondingAuthor":true,"prefix":"","firstName":"Mukendwa","middleName":"Hosticks","lastName":"Ndozi","suffix":""},{"id":619910233,"identity":"33a94751-d0c4-43d8-966d-1a5693cb5b58","order_by":1,"name":"Linnet Gohole","email":"","orcid":"","institution":"University of Eldoret","correspondingAuthor":false,"prefix":"","firstName":"Linnet","middleName":"","lastName":"Gohole","suffix":""},{"id":619910239,"identity":"cbd0536b-196d-4f89-857c-ed78a9589112","order_by":2,"name":"Isaac Mapaure","email":"","orcid":"","institution":"International University of Management","correspondingAuthor":false,"prefix":"","firstName":"Isaac","middleName":"","lastName":"Mapaure","suffix":""},{"id":619910241,"identity":"9d13442c-80db-4258-97cf-ba48ed977f0f","order_by":3,"name":"Emily Jepyegon Chemoiwa","email":"","orcid":"","institution":"University of Eldoret","correspondingAuthor":false,"prefix":"","firstName":"Emily","middleName":"Jepyegon","lastName":"Chemoiwa","suffix":""}],"badges":[],"createdAt":"2026-01-17 15:38:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8627044/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8627044/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106551530,"identity":"0b7ceeaf-a324-44ad-9ecf-7c2cabc7b506","added_by":"auto","created_at":"2026-04-09 18:24:19","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":545158,"visible":true,"origin":"","legend":"\u003cp\u003eStudy area showing the different sampling points in Nkasa Rupara National Park (Wildlife system) and Dzoti Conservancy (Wildlife-livestock system).\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-8627044/v1/626fe27420209284a4d8794e.png"},{"id":106551529,"identity":"58422e00-1933-43ed-a23b-1130d9029786","added_by":"auto","created_at":"2026-04-09 18:24:14","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":105289,"visible":true,"origin":"","legend":"\u003cp\u003eAgarose gel image of successful PCR amplification at 750 Base Pairs (bp), the first well representing the ladder while N is the code that represents the specimens from the national park and C represents the species that were loaded per well.\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-8627044/v1/c2b5148a8f392501f014e7e8.png"},{"id":106551553,"identity":"fb849a28-913a-4004-8961-3ff93ba13ff3","added_by":"auto","created_at":"2026-04-09 18:24:25","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":158194,"visible":true,"origin":"","legend":"\u003cp\u003eNeighbour Joining tree of the subfamily Scarabaeinae from wildlife and wildlife-livestock systems in north-eastern Namibia based on COI barcode sequences. Rooted with \u003cem\u003eEpirinus aeneus\u003c/em\u003eaccession number - GQ290002.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-8627044/v1/44fe3f92fe0f147fdca1bce8.png"},{"id":106551610,"identity":"8f3fb20a-3f43-4d5b-9bf7-35624ad540a8","added_by":"auto","created_at":"2026-04-09 18:24:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1788309,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8627044/v1/415e47a9-3dd2-483a-b0c0-a5728e74a5b0.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"DNA barcoding and phylogenetic analysis of dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae) in wildlife and wildlife-livestock systems of Namibia","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDung beetles are a group of insects that are regarded as coprophagous (mainly feeding on dung materials), and belong to the largest order of Coleoptera, family Scarabaeidae of the subfamily Scarabaeinae. The diversity of dung beetles is associated with the dung types excreted by mammals (Raine and Slade \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), vegetation types, characteristics of the soil and seasonality of rainfall (Tshikae et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). This group of insects are globally distributed with 442,275 described species in 29,595 genera (Goczał et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), with more than 7,000 described species belonging to the subfamily of Scarabaeinae (Daniel and Davis \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). About 540 species of dung beetles have been reported in the ecological zones of Southern African regions, especially in countries such as Botswana, Namibia and South Africa. Presently, 214 dung beetle species have been recorded in various ecosystems of Namibia, with 22 species have been identified as endemic species and majority of these endemic species are adapted to the Namib Desert (Davis et al. 2020). The presence of high species composition, diversity, abundance and assemblages of dung beetles in different ecosystem of Africa is primarily influenced by the presence of either abiotic or biotic factors (Daniel et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Rainfall patterns, temperature fluctuations, elevation and soil types are some of the abiotic factors that alters the assemblage structures of dung beetles (Davis et al. 2020; Daniel et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). On the other hand, some of the biotic factors that changes the assemblages of dung beetles includes habitat types, vegetation cover, dung resources that are influenced by the diversity and abundance of mammalian species (Pryke et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Bruda et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eDung beetles are sensitive to environmental changes that are inclined as a result of habitat modifications and anthropogenic actions, one of the reasons why these insects are primarily considered as bioindicators (Davis et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Nichols et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Environmental disturbances have been causing a serious replacement of native flora by large areas of agriculture and pasture (Nichols et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Loss in biodiversity as a result in the reduction of natural areas is the main leading factor to loss of genetic variability of the remaining species. Dung beetles are specialist in utilising dung pads for feeding, breeding and nesting (Heddle et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Thotagamuwa et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Several ecological functions and ecosystem services are complimented by dung beetles when utilising dung pads. The ecological processes that are driven by the dung beetles during the relocation and consumption of dung materials include the improvement of soil aeration, nutrient cycling, secondary seed burial, parasite suppression, bioturbation and fly control (Andresen \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Nichols et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Arellano et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMorphological identification of dung beetles remains a challenging task as a result of variable morphological differences among the species (Murphy 2020). Some of the morphological differences appears in different life stages of the species, for instance, it is somehow challenging to identify the larval stage of some insects. However, the identification keys on dung beetles using morphological characteristics are more effective on a particular life stage such as the adult stage (Bandral et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Jagdale et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Therefore, challenges related to the effective use of the taxonomic keys can be resolved with the use of DNA barcoding. The identification of dung beetles by the use of molecular markers is useful in eliminating inaccuracy and appears promising in solving the problem of misidentification of species (Johnston et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). DNA barcoding has different techniques that have been adopted for the identification and characterization of scarab beetles, which include the use of mitochondrial DNA (mtDNA)-encoded cytochrome oxidase I genes (COI) (Mlambo et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Baena-Bejarano et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Watanabe et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Villalba et al. (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2002\u003c/span\u003e) reported the first use of molecular identification in the subfamily of Scarabaeinae, whereas mitochondrial cytochrome oxidase I and II (COI, COII) were considered for PCR and DNA sequencing.\u003c/p\u003e \u003cp\u003eThe African continent is characterised by the best Scarabaeinae dung beetle diversity; a total of 2,000 species have been documented (Scholtz et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), 670 species inhabit Southern Africa, and 214 species have been reported in Namibia (Davis et al. 2020). Though DNA barcoding and phylogenetic analysis have been performed in Southern Africa (where Namibia is located), research using DNA to identify dung beetles has not been fully exploited in Namibia. Considering that the overall distribution and knowledge of DNA barcoding and phylogenetic analysis in the Namibian dung beetle community on a global scale are poor, the molecular study of the Scarabaeinae is still in need of treatment and recommendation of gene sequences to bridge this gap. The use of molecular data needs attention in order to address the problem of misidentification of each of these insects, focusing on species-level identification and the exploitation of molecular information to make a significant difference.\u003c/p\u003e \u003cp\u003eDung beetles are considered a cosmopolitan insect family with vital service roles in ecosystems (Davis and Scholtz 2001), and phylogenetic determination will help establish their phylogenetic positions and classify them into clades based on their tribal association. Since dung beetles of Namibia are under-investigated in molecular sequencing areas, this creates a significant gap in the global study of biogeography, distribution, and diversity of dung beetles. For example, Namibia has been able to document at most nine dung beetle tribes from morphological analysis (Davis et al. 2020; Ndozi et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2025\u003c/span\u003ea). In a similar manner, the relationship of evolution, as reflected in the Bayesian phylogram, showed that there are nine African Scarabaeinae tribes, which include Ateuchini, Coprini, Canthonini, Gymnopleurini, Oniticellini, Onitini, Onthophagini, Scarabaeini, and Sisyphini. This phylogenetic relationship is critical in the study of dung beetles and also in understanding genetic flow, identification, and population organization. Therefore, the study determined the DNA barcoding and phylogenetic analysis of the dung beetles in wildlife and wildlife-livestock systems of Namibia. Thus, phylogeographic studies and biodiversity assessments are therefore supported by the correct species identification, phylogenetic position of the species, and the availability of the DNA barcodes. Such kind of information has not documented with reference to Namibian dung beetles.\u003c/p\u003e"},{"header":"Methods and Materials","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy area\u003c/h2\u003e \u003cp\u003eThe study was conducted at Nkasa Rupara National Park (23.607676 E; 18.364642 S), which is a wildlife system, and Dzoti Conservancy (23.781766 E; 18.252163 S), which is the wildlife-livestock system (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Both study sites are situated in the north-eastern part of Namibia. The selection of the two systems was mainly based on the similarities of the vegetation, soil type, and rainfall seasonality. The national park is characterised by the riparian woodland and grassland that mostly dominant in floodplain areas, and the park covers a distance of 900 km\u003csup\u003e2\u003c/sup\u003e (MEFT, 2020). While Dzoti Conservancy is a wildlife-livestock system covering an area of 287 km2 and is home to an estimated human population of over 2,000 (Denker \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Woodland vegetation is mainly dominated by the broadleaved trees including tree species such as \u003cem\u003eCombretum imberbe\u003c/em\u003e (Wawra 1860), \u003cem\u003eKigelia africana\u003c/em\u003e (Lamarck 1849), \u003cem\u003eSenegalia nigrescens\u003c/em\u003e (Oliver et al. 1871), \u003cem\u003eVachellia sieberiana\u003c/em\u003e (de Candolle 1825), \u003cem\u003eAlbizia versicolour\u003c/em\u003e (Oliver 1871), \u003cem\u003ePiliostigma thonningii\u003c/em\u003e (Schumach 1947), and \u003cem\u003eTerminalia sericea\u003c/em\u003e (Burchell 1828). The floodplain areas are dominated by the grass species of \u003cem\u003eHyparrhenia rufa\u003c/em\u003e (Stapf 1919), other non-woody species adapted in floodplains includes reeds, edges and wild date palms. The vegetation types along the riverbanks and channels are dominated by broadleaved trees such as \u003cem\u003eDiospyros mespiliformis\u003c/em\u003e (Hochstetter 1844) and \u003cem\u003eGarcinia livingstonei\u003c/em\u003e (Anderson1866). The mammalian species that are common in the national park includes the cape buffalo, elephant, wildebeest, hippopotamus and several others species. The cambisol soil types dominates national park (FAO and ITPS 2015). Rain begins to fall in late November and lasts until mid-March, with an annual rainfall ranging from 400\u0026ndash;800 mm and temperatures ranging from 24\u0026ndash;28\u0026deg;C. In winter, minimum temperature ranges between 2\u0026deg;C and 8\u0026deg;C, while in summer, maximum temperature can range between 36\u0026deg;C and 40\u0026deg;C (Ndozi et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2025\u003c/span\u003ea; \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2025\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSampling of dung beetles\u003c/h3\u003e\n\u003cp\u003eDung beetles were sampled either by hand picking from the abundant bovine dung pads, whereas some of the dung beetles were collected using pitfall traps. For small (less than 2 mm) dung beetle species, the entire insect was placed in a 2 ml specimen vial with DNA Shield solution, with a maximum of 4 individuals placed in each vial. Dung beetles that had a lot of dung materials on their exoskeletons were either excluded or washed with deionized water to remove the excrements. In large specimens (2 mm \u0026ndash; 5 mm), the middle femurs were removed, then sliced into small pieces and placed in a labelled specimen vial. Other parts such as the abdomen, head and thorax were excluded in larger species as they may contain dung materials that might interfere with the process of DNA extraction.\u003c/p\u003e\n\u003ch3\u003ePreservation of specimens\u003c/h3\u003e\n\u003cp\u003eAll the dung beetle species that were collected from both systems were preserved in DNA/RNA shield (Zymo Research). After placing the samples in DNA/RNA shield, samples were kept in a freezer (-20 ℃) before the initial transportation. All the collected samples were transported to the Fish Genomic and Genetic Research Laboratory (FGGRL). At the Fish Genomic and Genetic Research Laboratory (FGGRL), University of Eldoret, samples were transferred to the freezer at -20 ℃ temperature. The DNA extraction and PCR amplification was conducted at the FGGRL.\u003c/p\u003e\n\u003ch3\u003ePretreatment of samples and DNA extraction\u003c/h3\u003e\n\u003cp\u003eDNA from each species of dung beetle was extracted separately using the blood and tissue Quick-DNA Miniprep Plus Kit (Zymo Research) according to the protocols of the manufacturer in exception of few modifications as described below. Prior to the extraction, samples (30 mg) were placed in 1.5 ml eppendorf tubes followed by the addition of warm double-distilled water (30\u0026deg;C), and vortexing was done for 15 seconds. After three times of washing, samples were crushed in the eppendorf tubes using Eppendorf micropestles.\u003c/p\u003e\n\u003ch3\u003eDNA quantification\u003c/h3\u003e\n\u003cp\u003eThe extracted DNA was quantified using the Nanodrop spectrophotometer (Thermo Fisher Scientific) to verify the concentration (ng/\u0026micro;l) of each DNA sample. The determination of purity of DNA was performed by taking the ratio of the sample absorbance at 260 and 230 nm (260/230) and also 260 and 280 nm (260/280).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003ePolymerase Chain Reaction (PCR) amplification\u003c/h2\u003e \u003cp\u003eThe amplifications were carried out with mtDNA using two sets of universal primers based on the cytochrome c oxidase gene I (COI) barcodes (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The PCR reactions were carried out in clear 0.2 ml microtubes with caps. The total PCR reaction volume of 25 \u0026micro;l was used: 6.5 \u0026micro;l was nucleic acid-free water; 12.5 \u0026micro;l was master mix; 0.5 \u0026micro;l was the forward primer; 0.5 \u0026micro;l was the reverse primer; and 5 \u0026micro;l consisted of template DNA (Asha and Sinu \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). mtDNA cytochrome c oxidase gene I (COI) barcoding universal primers were added: COI forward primer (C1-J-2183): 5\u0026rsquo;CAACATTTATTTTGATTTTTTGG-3\u0026rsquo; and COI reverse primer (TL2-N-3014): 5\u0026rsquo;TCCAATGCACTAATCTGCCATATTA-3\u0026rsquo; (Simon et al. 1994) were used. PCR amplification was done using the following conditions: The initial denaturation temperature was set at 94\u0026deg;C for 90 seconds, followed by denaturation at 94\u0026deg;C for 22 seconds, followed by 35 cycles, annealing at 50\u0026deg;C for 30 seconds, extension at 72\u0026deg;C for 90 seconds, and the final extension was at 72\u0026deg;C for 60 seconds (Mlambo et al. 2011). For the second primers, the PCR reaction was performed following the established protocol similar to the first primers using COI forward primer (LCO1490): 5\u0026rsquo;GGTCAACAAATCATAAAGATATTGG-3\u0026rsquo; and COI reverse primer (HCO2198): 5\u0026rsquo;TAAACTTCAGGGTGACCAAAAAATCA-3\u0026rsquo; (Folmer et al. 1994). PCR amplification was done using the following conditions: The initial denaturation temperature was set at 95\u0026deg;C for 120 seconds, followed by denaturation at 95\u0026deg;C for 30 seconds, followed by 34 cycles, annealing at 50\u0026deg;C for 60 seconds, extension at 72\u0026deg;C for 2 minutes, and the final extension was at 72\u0026deg;C for 7 minutes (Gavarāne et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The amplicons were confirmed by the gel electrophoresis using 1% agarose (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The amplified products were then sent to Inqaba Biotec East Africa (Ltd) in Nairobi, where sequencing was done.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eA total of 62 sequences of 740 base pairs (bp) representing 33 species of 23 genera (\u003cem\u003eAllogymnopleurus\u003c/em\u003e, \u003cem\u003eCaccobius\u003c/em\u003e, \u003cem\u003eCatharsius\u003c/em\u003e, \u003cem\u003eChalconotus\u003c/em\u003e, \u003cem\u003eCopris\u003c/em\u003e, \u003cem\u003eDigitonthophagus\u003c/em\u003e, \u003cem\u003eEuonthophagus\u003c/em\u003e, \u003cem\u003eGymnopleurus\u003c/em\u003e, \u003cem\u003eKheper\u003c/em\u003e, \u003cem\u003eKurtops\u003c/em\u003e, \u003cem\u003eLatodrepanus\u003c/em\u003e, \u003cem\u003eMetacatharsius\u003c/em\u003e, \u003cem\u003eMilichus\u003c/em\u003e, \u003cem\u003eOniticellus\u003c/em\u003e, \u003cem\u003eOnitis\u003c/em\u003e, \u003cem\u003eOnthophagus\u003c/em\u003e, \u003cem\u003ePachylomera\u003c/em\u003e, \u003cem\u003ePhalops\u003c/em\u003e, \u003cem\u003eScarabaeus\u003c/em\u003e, \u003cem\u003eSisyphus\u003c/em\u003e, and \u003cem\u003eTiniocellus\u003c/em\u003e) were taken along with 1 species of \u003cem\u003eEpirinus aeneus\u003c/em\u003e (GQ290002) using the COI gene fragment for phylogenetic analysis belonging to the subfamily of Scarabaeinae. The species \u003cem\u003eE. aeneus\u003c/em\u003e of tribe Deltochilini of the same subfamily was used as an outroot for using the COI gene fragment for phylogenetic analysis. The species \u003cem\u003eE. aeneus\u003c/em\u003e of the subfamily Scarabaeinae is regarded to be an endemic species of southern Africa. The construction of the phylogenetic tree was based on COI gene sequences using 1,000 bootstrapped Neighbour-Joining with the Kimura-2 parameters.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eData analysis\u003c/h2\u003e \u003cp\u003eThe sequence FASTA files were aligned using multiple alignments ClustalW software and edited using the BioEdit Sequence Alignment Editors v 7.2; this was performed to eliminate components that dispersed and bases that were unclear. After aligning and trimming the sequences, their length was 786bp. The similarities of the sequences were aligned using the BLAST algorithm, and a database of 62 sequences was created. The collection details of all the sequences that were uploaded to the Bold Systems and accession numbers recorded. The MEGA software version 12.0 (Kumar et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) was used in performing the phylogenetic relationships by Neighbor Joining (NJ) analysis whereby Kimura\u0026rsquo;s two-parameter substitution model was used, with a consideration of 1,000 bootstrap replicates (Tamura et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). FigTree version 1.4.4 was used to visualize the phylogenetic tree.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eDNA barcoding\u003c/h2\u003e \u003cp\u003eAll the sequences for wildlife and wildlife-livestock systems samples were compared by performing BLAST analysis with those sequences in the NCBI GenBank. The present study compared the barcode of 61 mtDNA amplicons of the genera \u003cem\u003eAllogymnopleurus\u003c/em\u003e, \u003cem\u003eCaccobius\u003c/em\u003e, \u003cem\u003eCatharsius\u003c/em\u003e, \u003cem\u003eChalconotus\u003c/em\u003e, \u003cem\u003eEuoniticellus\u003c/em\u003e, \u003cem\u003eEuonthophagus\u003c/em\u003e, \u003cem\u003eGymnopleurus\u003c/em\u003e, \u003cem\u003eHeliocopris\u003c/em\u003e, \u003cem\u003eKheper\u003c/em\u003e, \u003cem\u003eLatodrepanus\u003c/em\u003e, \u003cem\u003eMetacatharsius\u003c/em\u003e, \u003cem\u003eMilichus\u003c/em\u003e, \u003cem\u003eOnitis\u003c/em\u003e, \u003cem\u003eOnthophagus\u003c/em\u003e, \u003cem\u003ePachylomera\u003c/em\u003e, \u003cem\u003ePhalops\u003c/em\u003e, \u003cem\u003eScarabaeus\u003c/em\u003e, \u003cem\u003eSisyphus\u003c/em\u003e, and \u003cem\u003eTiniocellus\u003c/em\u003e dung beetles submitted to Bold Systems. The respective species were identified to genus and species level. From a total of 27 DNA sequences which matched in the NCBI GenBank, 15 species were identified with the percentage of 95\u0026ndash;100% identity distributed in to 14 genera. In the 27 sequences identified, twelve species appeared common from the two study ecosystems in Namibia (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Approximately 34 dung beetle species identified morphologically as earlier reported by Ndozi et al. (2024) were not retrieved and identified from the NCBI database (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The unretrieved were 24 species belonging to 11 genera showing a high population of Namibian dung beetles as being under reported.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eScarabaeinae taxa studied in wildlife and wildlife-livestock ecosystems and their Bold Systems accession numbers, percent identity and COI primers.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLab ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpecies name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBOLD systems Accession Numbers\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePrimer set used\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePercent identity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMatched Accession\u003c/p\u003e \u003cp\u003eNumbers in NCBI\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eAllogymnopleurus thalassinus\u003c/em\u003e (Klug,1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC020\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131898.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLatodrepanus laticollis\u003c/em\u003e (Fahraeus, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEF188137.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMetacatharsius opacus\u003c/em\u003e (Waterhouse, 1891)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131865.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMilichus apicalis\u003c/em\u003e (Fahraeus, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131921\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnitis alexis\u003c/em\u003e (Klug, 1835)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC017\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHCO2198/LCO1490\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHM375977.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus variegatus\u003c/em\u003e (Erichson, 1843)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEF188218\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePachylomera femoralis\u003c/em\u003e (Kirby, 1828)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC007\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eJN804658.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eSisyphus goryi\u003c/em\u003e Harold, 1859\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMH129898.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eChalconotus convexus\u003c/em\u003e (Boheman, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131809.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eTiniocellus spinipes\u003c/em\u003e (Roth, 1851)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131912.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eScarabaeus zambesianus\u003c/em\u003e P\u0026eacute;ringuey, 1901\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAF499770.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eDigitonthophagus gazella\u003c/em\u003e (Fabricius, 1787)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131918.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEuonthophagus carbonarius\u003c/em\u003e (Klug,1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131919\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eAllogymnopleurus thalassinus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC018\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131898.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEuonthophagus carbonarius\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131919\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKurtops signatus\u003c/em\u003e (Fahraeus, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEF188216\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLatodrepanus laticollis\u003c/em\u003e (Fahraeus, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC022\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEF188137.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMetacatharsius opacus\u003c/em\u003e (Waterhouse, 1891)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131865.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMilichus apicalis\u003c/em\u003e (Fahraeus, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131921.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnitis alexis\u003c/em\u003e (Klug, 1835)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC027\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHCO2198/LCO1490\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eKF801856.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus fimetarius\u003c/em\u003e (Roth, 1851)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131925\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus variegatus\u003c/em\u003e (Erichson, 1843)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC021\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEF188217\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePachylomera femoralis\u003c/em\u003e (Kirby, 1828)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC008\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eJN804658.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eSisyphus goryi\u003c/em\u003e Harold, 1859\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC028\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMH129898.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eTiniocellus spinipes\u003c/em\u003e (Roth, 1851)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131912.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eScarabaeus zambesianus\u003c/em\u003e P\u0026eacute;ringuey, 1901\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC009\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAF499770.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eDigitonthophagus gazella\u003c/em\u003e (Fabricius, 1787)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAY131918\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eUnretrieved dung beetle sequences in gene bank database and BOLD systems.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLa ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpecies name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBOLD systems Accession Numbers\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePrimer set used\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKheper lamarcki\u003c/em\u003e (Macleay, 1821)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKheper nigroaeneus\u003c/em\u003e (Boheman, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC030\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnitis deceptor\u003c/em\u003e (P\u0026eacute;ringuey, 1901)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC031\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCatharsius aegeus\u003c/em\u003e (G\u0026eacute;nier, 2017)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus lamelliger\u003c/em\u003e (Gerstaecker, 1871)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC033\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus quadrinotatus\u003c/em\u003e (d'Orbigny, 1905)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus vinctus\u003c/em\u003e (Erichson,1843)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC035\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePhalops flavocinctus\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC036\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKheper prodigiosus\u003c/em\u003e (Erichson, 1843)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC037\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eHeliocopris japetus\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC038\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCatharsius tricornutus\u003c/em\u003e (DeGeer, 1778)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCopris amyntor\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCopris elphenor\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC040\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCopris puncticollis\u003c/em\u003e (Boheman, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC041\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eGymnopleurus pumilus\u003c/em\u003e (Reiche, 1850)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC061\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHCO2198/ LCO1490\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEuoniticellus intermedius\u003c/em\u003e (Reiche, 1849)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC059\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHCO2198/ LCO1490\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eGymnopleurus pumilus\u003c/em\u003e (Reiche,1850)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC042\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHCO2198/LCO1490\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKheper lamarcki\u003c/em\u003e (Macleay, 1821)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC060\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKheper nigroaeneus\u003c/em\u003e (Boheman, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC043\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCaccobius nigritulus\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC044\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOniticellus formosus\u003c/em\u003e (Chevrolat, 1830)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC045\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnitis deceptor\u003c/em\u003e (P\u0026eacute;ringuey, 1901)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC046\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus aeruginosus\u003c/em\u003e (Roth, 1851)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC047\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus flavolimbatus\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC048\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus lamelliger\u003c/em\u003e (Gerstaecker, 1871)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC049\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCatharsius aegeus\u003c/em\u003e (G\u0026eacute;nier, 2017)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC050\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus quadrinotatus\u003c/em\u003e (d'Orbigny, 1905)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC051\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus suffusus\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC052\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOnthophagus vinctus\u003c/em\u003e (Erichson, 1843)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC053\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePhalops boschas\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC054\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePhalops flavocinctus\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eKheper prodigiosus\u003c/em\u003e (Erichson, 1843)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC056\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCopris elphenor\u003c/em\u003e (Klug, 1855)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC057\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCopris puncticollis\u003c/em\u003e (Boheman, 1857)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNAMC058\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTL2N3014/C1J2183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic analysis of dung beetles\u003c/h2\u003e \u003cp\u003eThe Neighbour-Joining phylogenetic tree (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) showed that 9 species of the genera \u003cem\u003eOnthophagus\u003c/em\u003e, \u003cem\u003eMilichus\u003c/em\u003e, and \u003cem\u003eCaccobius\u003c/em\u003e of the tribe Onthophagini separated from each other and grouped together with the bootstrap value (\u0026gt;\u0026thinsp;95%). \u003cem\u003eDigitonthophagus gazella\u003c/em\u003e, \u003cem\u003eEuonthophagus carbonarius\u003c/em\u003e, \u003cem\u003eKurtops signatus\u003c/em\u003e and \u003cem\u003eOnthophagus flavolimbatus\u003c/em\u003e of the similar tribe Onthophagini also separated and grouped together with the bootstrap value (\u0026gt;\u0026thinsp;95%). The clustering examination divided the 32 species into three main clades, while two species (NP37 \u003cem\u003ePhalops flavocinctus\u003c/em\u003e and NP21 \u003cem\u003eOniticellus formosus)\u003c/em\u003e appeared separately from the three clades. The first clade consisted mainly of species from the genera \u003cem\u003eOnthophagus\u003c/em\u003e, \u003cem\u003eMilichus\u003c/em\u003e, \u003cem\u003eCaccobius\u003c/em\u003e, \u003cem\u003eLatodrepanus\u003c/em\u003e, \u003cem\u003eOnitis\u003c/em\u003e, \u003cem\u003eSisyphus\u003c/em\u003e, \u003cem\u003eTiniocellus\u003c/em\u003e, \u003cem\u003eKurtops\u003c/em\u003e, \u003cem\u003eEuonthophagus\u003c/em\u003e, and \u003cem\u003eDigitonthophagus\u003c/em\u003e. The second clade was composed of species of the genera \u003cem\u003eAllogymnopleurus\u003c/em\u003e, \u003cem\u003eGymnopleurus\u003c/em\u003e, \u003cem\u003eCopris\u003c/em\u003e, \u003cem\u003eMetacatharsius\u003c/em\u003e, \u003cem\u003ePhalops\u003c/em\u003e, \u003cem\u003ePachylomera\u003c/em\u003e, \u003cem\u003eKheper\u003c/em\u003e, and \u003cem\u003eScarabaeus\u003c/em\u003e. The third clade included two species: \u003cem\u003eCatharsius aegeus\u003c/em\u003e and \u003cem\u003eCatharsius tricornutus\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study provided the DNA barcodes of 62 species of the genera \u003cem\u003eAllogymnopleurus\u003c/em\u003e, \u003cem\u003eCaccobius\u003c/em\u003e, \u003cem\u003eCatharsius\u003c/em\u003e, \u003cem\u003eChalconotus\u003c/em\u003e, \u003cem\u003eEuoniticellus\u003c/em\u003e, \u003cem\u003eEuonthophagus\u003c/em\u003e, \u003cem\u003eGymnopleurus\u003c/em\u003e, \u003cem\u003eHeliocopris\u003c/em\u003e, \u003cem\u003eKheper\u003c/em\u003e, \u003cem\u003eLatodrepanus\u003c/em\u003e, \u003cem\u003eMetacatharsius\u003c/em\u003e, \u003cem\u003eMilichus\u003c/em\u003e, \u003cem\u003eOnitis\u003c/em\u003e, \u003cem\u003eOnthophagus\u003c/em\u003e, \u003cem\u003ePachylomera\u003c/em\u003e, \u003cem\u003ePhalops\u003c/em\u003e, \u003cem\u003eScarabaeus\u003c/em\u003e, \u003cem\u003eSisyphus\u003c/em\u003e, and \u003cem\u003eTiniocellus\u003c/em\u003e dung beetles. Dung beetles from different ecological zones of Namibia are poorly documented in the GenBank data base such as the BOLD systems and NCBI, hence some of the sequences (a total of 35 dung beetle species) were not identified to their respective species level However, the woodland savanna and mixed grassland ecosystems spanning the entire north-eastern part of Namibia are less explored for invertebrates including arthropods, despite having favorable microclimates that might support the species diversity and richness of different insect communities. The distribution and phylogenetic origin of dung beetles in both wildlife and wildlife-livestock systems appear to be unclear, contributing to the poor representation of dung beetles of Namibia in the global DNA barcoding, phylogenetic and biogeographical investigations and therefore the GenBank data base.\u003c/p\u003e \u003cp\u003eAround 85% of the BLAST sequences compared were similar to dung beetles from the Sub-Saharan biogeographical region. Most of the sequences found in wildlife and wildlife-livestock systems did not have reference sequences publicly entered into the database. The current study is the first report by researchers of DNA barcodes for Namibian dung beetles. Little has been done regarding molecular identification, especially in the tribe of Onthophagini. \u003cem\u003eOnthophagus flavolimbatus\u003c/em\u003e Klug, 1855, for example, one of the species from this tribe first appeared in the wildlife ecosystem of the northeastern part of Namibia. However, South Africa (Tocco et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and Mozambique (Daniel and G\u0026eacute;nier \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) reported this species to be present in parts of these countries, even though there was no DNA sequence for it in this database. The tribe Onthophagini, to which \u003cem\u003eO. flavolimbatus\u003c/em\u003e belongs, as well as other tribes like Sisyphini, Oniticellini, and Coprini, are considered cosmopolitan (Davis and Scholtz 2001). Apart from \u003cem\u003eO. flavolimbatus\u003c/em\u003e, the study further provided insights on the first mitochondrial cytochrome c oxidase subunit 1 (COI) gene sequences of the species, including \u003cem\u003eCopris elphenor\u003c/em\u003e Klug 1855, \u003cem\u003eHeliocopris japetus\u003c/em\u003e Klug, 1855, \u003cem\u003eKheper lamarcki\u003c/em\u003e Macleay 1821, \u003cem\u003eOniticellus formosus\u003c/em\u003e Chevrolat 1830, \u003cem\u003eOnthophagus aeruginosus\u003c/em\u003e Roth 1851, \u003cem\u003eOnthophagus lamelliger\u003c/em\u003e Gerstaecker 1871, \u003cem\u003eOnthophagus vinctus\u003c/em\u003e Erichson 1843, and \u003cem\u003ePhalops flavocinctus\u003c/em\u003e Klug 1855. The DNA barcodes of these species have not yet been uploaded in the BOLD systems and GenBank of the NCBI database, making it difficult to identify these species using the molecular marker.\u003c/p\u003e \u003cp\u003eThere are three genera that represents tribe Gymnopleurini in Africa (Davis et al. 2020), however, the present study only captured two of the genera belonging to this tribe, namely \u003cem\u003eAllogymnopleurus\u003c/em\u003e and \u003cem\u003eGymnopleurus\u003c/em\u003e. The findings of this study did not report on genus \u003cem\u003eGarreta\u003c/em\u003e, which is the third genus in the tribe Gymnopleurini. Phylogenetic analysis also revealed that Gymnopleurini is a sister group to Scarabaeini, which indicates that this tribe is more closely related to the genus \u003cem\u003ePachylomera\u003c/em\u003e in that sister group. Based on phylogenetic analyses carried out elsewhere, the tribe has also been placed in a similar clade to the tribe Scarabaeini, while phylogenetic studies from other sources also show that the tribe Gymnopleurini is in a clade with Scarabaeini, hence demonstrating a strong sister relationship (Villalba et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Monaghan et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Mlambo et al. 2011). \u003cem\u003eGymnopleurini pumilus\u003c/em\u003e Reiche 1850, was not found in the database (unlike the species \u003cem\u003eAllogymnopleurus thalassinus\u003c/em\u003e Klug 1855), indicating insufficient records on the phylogenetic work of the tribe Gymnopleurini. A relationship was revealed in the dung beetle species of \u003cem\u003eMetacatharsius opacus\u003c/em\u003e Waterhouse 1891, within the tribe of Coprini and species from the tribe Sisyphini (\u003cem\u003eSisyphus goryi\u003c/em\u003e Harold 1859). Philips et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) and Mlambo et al. (2011) displayed a similar phylogenetic relationship in species of the genus \u003cem\u003eMetacatharsius\u003c/em\u003e. In addition, the species from the tribe Coprini are regarded as tunnelers, while those species in the tribe of Sisyphini are representing all the rollers.\u003c/p\u003e \u003cp\u003eA monophyletic lineage was noticed in the genus of \u003cem\u003eOnitis\u003c/em\u003e, showing a close relationship with the dung beetle species from the genera \u003cem\u003eTiniocellus\u003c/em\u003e and \u003cem\u003eLatodrepanus\u003c/em\u003e. Additionally, tribe Oniticellini and Onthophagini, both showed a monophyletic lineage to some of the species within the tribe of Onitini. Mlambo et al. (2011) explained that the genera of \u003cem\u003eCheironitis\u003c/em\u003e and \u003cem\u003eOnitis\u003c/em\u003e, all from the tribe Onitini, indicated a sister relationship with majority of the species from the tribe Onthophagini. According to Villalba et al. (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2002\u003c/span\u003e) discovered an exciting understanding of the sisterhood relationship between the tribes Onitini and Onthophagini. Several molecular phylogeny studies have revealed the sister relationship of tribe Onitini are closely related to tribes Onthophagini and Oniticellini (Villalba et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Monaghan et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). In addition, Philips et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) indicated that Onthophagini (\u003cem\u003eDigitonthophagus\u003c/em\u003e), Onitini (\u003cem\u003eBabas\u003c/em\u003e, \u003cem\u003eHeteronitis\u003c/em\u003e and \u003cem\u003eOnitis\u003c/em\u003e), and Oniticellini (\u003cem\u003eTiniocellus\u003c/em\u003e) were in one clade, showing the sister relationship.\u003c/p\u003e \u003cp\u003eDung beetles that belong to the tribe Onthophagini showed paraphyletic grouping; for instance, the species \u003cem\u003eDigitonthophagus gazella\u003c/em\u003e Fabricius 1787, \u003cem\u003eEuonthophagus carbonarius\u003c/em\u003e Klug 1855, \u003cem\u003eKurtops signatus\u003c/em\u003e Fahraeus 1857, and \u003cem\u003eOnthophagus flavolimbatus\u003c/em\u003e Klug 1855, formed a clade that is sister to the rest of the genera (Villalba et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Breeschoten et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Philips et al. 2016; Zhang et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). In addition, the paraphyletic grouping within the tribe Onthophagini is due to presence of tribe Oniticellini. The results of this study revealed inconsistency of the genus \u003cem\u003eOnthophagus\u003c/em\u003e within the tribe Onthophagini in their evolutionary clade. \u003cem\u003eOnthophagus\u003c/em\u003e is presently categorized as highly polyphyletic among all the genus within the tribe of Onthophagini. Species such as \u003cem\u003eCaccobius nigritulus\u003c/em\u003e, and \u003cem\u003eMilichus apicalis\u003c/em\u003e were nested within the genus \u003cem\u003eOnthophagus\u003c/em\u003e, despite the two genera been scarcely defined. The analysis confirmed the polyphyletic status of tribe Onthophagini within the main clade, which might have been influenced by the rearrangement among the terminal taxa, without changing their relationships and main lineage. Tribe Coprini was represented mainly by three genera (\u003cem\u003eCatharsius\u003c/em\u003e, \u003cem\u003eCopris\u003c/em\u003e, and \u003cem\u003eMetacatharsius\u003c/em\u003e); species from the two genera, \u003cem\u003eCopris\u003c/em\u003e and \u003cem\u003eMetacatharsius\u003c/em\u003e, appeared to be a sister taxon, sharing suitable synapomorphies that might accurately define the tribe Coprini. In contrast, the species that belongs to \u003cem\u003eCatharsius\u003c/em\u003e showed a homoplasy (Santis \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) that they do not share a common ancestral root, despite being classified within the same tribe of Coprini.\u003c/p\u003e \u003cp\u003eThe species \u003cem\u003ePachylomera femoralis\u003c/em\u003e Kirby 1828, the only species representing the genus \u003cem\u003ePachylomera\u003c/em\u003e from the tribe Scarabaeini demonstrated a close relationship to \u003cem\u003eScarabaeus zambesianus\u003c/em\u003e P\u0026eacute;ringuey 1901. Both \u003cem\u003ePachylomera femoralis\u003c/em\u003e and \u003cem\u003eScarabaeus zambesianus\u003c/em\u003e are from the same tribe. Taxonomists have been proposing that the species \u003cem\u003ePachylomera femoralis\u003c/em\u003e should be a subgenus to \u003cem\u003eScarabaeus\u003c/em\u003e (Harrison et al. 2003; Forgie et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), while other studies have supported the current genus of \u003cem\u003ePachylomera\u003c/em\u003e (Scholtz et al. 2011). This study fully supports the classification of \u003cem\u003ePachylomera\u003c/em\u003e as a genus. Furthermore, \u003cem\u003eKheper lamarcki\u003c/em\u003e Macleay 1821, \u003cem\u003eKheper nigroaeneus\u003c/em\u003e Boheman 1857, and \u003cem\u003eKheper prodigiosus\u003c/em\u003e Erichson 1843, the only three species representing the genus \u003cem\u003eKheper\u003c/em\u003e exhibited a sister relationship with \u003cem\u003eScarabaeus zambesianus\u003c/em\u003e in this study confirming its classification as a genus. The results obtained by Mlambo et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) showed a close grouping of the species from the genera \u003cem\u003eKheper\u003c/em\u003e and \u003cem\u003escarabaeus\u003c/em\u003e, supporting their classification as separate genera. Hence, all the dung beetles belonging to the three genera (\u003cem\u003eKheper\u003c/em\u003e, \u003cem\u003ePachylomera\u003c/em\u003e, and \u003cem\u003eScarabaeus\u003c/em\u003e) of the tribe Scarabaeini are regarded as rollers. Rollers are group of dung beetles with the nesting behaviours of forming a brood ball that is pushed a distance from the dung pad and buried into the soil. The use of the COI gene can effectively disparate the numerous scarabs of Namibia, including the scarabs of southern Africa and other species that are found elsewhere in the world. Therefore, the application of molecular identification in dung beetles is valuable for those species that are challenging to be identified using morphologic characters and assists in identifying the unidentified dung beetle species.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eDNA barcoding and phylogenetic analysis of dung beetles in wildlife and wildlife-livestock systems provided accurate identification at the species level, creating opportunities for future investigations. The use of the COI gene for species identification, which was adopted for this study, remains the technique for identifying species-level characteristics. Furthermore, the study complements mitochondrial COI sequences of several species of dung beetles to the database, thereby making these mitochondrial barcodes available for further studies in the same field. The COI gene is recommended as a more effective marker for DNA barcoding and phylogenetic analysis in dung beetle species; however, conducting a comprehensive investigation of population genetics using other genes is essential to confirm the species that have appeared and been identified morphologically in Namibia for the first time.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe authors (Mukendwa Hosticks Ndozi, Linnet Gohole, Isaac Mapaure, Emily Jepyegon Chemoiwa) state and declare that there are no competing interests and this work has not been published elsewhere.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the University of Eldoret through the Fishery Genomics and Genetic Laboratory for allowing us to conduct the DNA extraction and PCR amplification. Many thanks goes to Inqaba East Africa, who assisted with sequencing and other helpful support.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis manuscript was prepared through a collective effort between the listed co-authors; all the listed authors contributed to the research design, collection of data, data analysis, literature review, and revision of the manuscript. All the listed authors played a significant role in completing this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was funded by the University of Namibia under the office of the Pro-Vice Chancellor- Research, Innovation and Development via a scholarship awarded to the lead author (Mukendwa Hosticks Ndozi).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAndresen E (2002) Dung beetles in a Central Amazonian rainforest and their ecological role as secondary seed dispersers. 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Ecol Evol, 15(8), e71906\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-tropical-insect-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtis","sideBox":"Learn more about [International Journal of Tropical Insect Science](http://link.springer.com/journal/42690)","snPcode":"42690","submissionUrl":"https://www.editorialmanager.com/jtis/default2.aspx","title":"International Journal of Tropical Insect Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Scarabaeinae, Molecular identification, COI, barcodes","lastPublishedDoi":"10.21203/rs.3.rs-8627044/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8627044/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTaxonomical keys have been used in the identification of dung beetle species due to its cost effectiveness and accessibilities. However, morphological identification of dung beetles remains a challenging task as a result of variable morphological differences among the species. The identification of dung beetles by the use of molecular markers helps in avoiding inaccuracy and appears promising to solve the problem of identifying species. The aim of this study was to determine the DNA barcoding and phylogenetic analysis of dung beetles in wildlife and wildlife-livestock ecosystems of Namibia. Sampling of dung beetles was done using baited pitfall traps and some species were handpicked from the abundant bovine dung pads. The collected dung beetles were preserved in DNA/RNA shield (Zymo Research), and kept in a freezer (-20 ℃) before the initial transportation. The genomic DNA was isolated using blood and tissue Quick-DNA Miniprep Plus Kit (Zymo Research). Two sets of universal primers based on the cytochrome c oxidase gene I (COI) barcodes selected for use. The sequence FASTA files were aligned using multiple alignments ClustalW software, and the phylogenetic relationships by Neighbor joining analysis was performed using the MEGA12 software. A total of 62 DNA barcodes of dung beetle species were accomplished from which 15 species from 27 sequenced samples were identified with the percentage of 95\u0026ndash;100%, while 35 dung beetle sequences were not retrieved from the database. Indicating that most of the sequences found in wildlife and wildlife-livestock systems did not have reference sequences publicly entered into the database. The Neighbour-Joining phylogenetic tree showed 9 species of the genera \u003cem\u003eOnthophagus\u003c/em\u003e, \u003cem\u003eMilichus\u003c/em\u003e, and \u003cem\u003eCaccobius\u003c/em\u003e of the tribe Onthophagini separated from each other and grouped together with the bootstrap value (\u0026gt;\u0026thinsp;95%). The clustering examination divided the 32 species into three main clades, while two species appeared separately from the three clades. This constituted the first report of DNA barcodes for Namibian dung beetles. The DNA barcoding and phylogenetic analysis application for molecular identification in dung beetles provides valuable and accurate information, which is useful in understanding their diversity.\u003c/p\u003e","manuscriptTitle":"DNA barcoding and phylogenetic analysis of dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae) in wildlife and wildlife-livestock systems of Namibia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-09 18:23:27","doi":"10.21203/rs.3.rs-8627044/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-05-16T19:41:14+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-05-15T09:22:13+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-05-07T15:17:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"208080523503860111116463327885127800060","date":"2026-04-24T22:03:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"40622320387120680026593535280434726453","date":"2026-04-22T13:21:53+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-08T22:16:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"107149304542086253817589814986916796912","date":"2026-04-06T16:03:14+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-05T14:30:26+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-21T07:28:24+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-21T07:24:47+00:00","index":"","fulltext":""},{"type":"submitted","content":"International Journal of Tropical Insect Science","date":"2026-01-17T15:25:32+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-tropical-insect-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtis","sideBox":"Learn more about [International Journal of Tropical Insect Science](http://link.springer.com/journal/42690)","snPcode":"42690","submissionUrl":"https://www.editorialmanager.com/jtis/default2.aspx","title":"International Journal of Tropical Insect Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"61a74375-a773-40cd-94b2-79349358b4e5","owner":[],"postedDate":"April 9th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Revision requested","date":"2026-05-16T19:41:14+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-05-15T09:22:13+00:00","index":47,"fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-05-07T15:17:39+00:00","index":46,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-05-16T19:53:45+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-09 18:23:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8627044","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8627044","identity":"rs-8627044","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}