Quarantine Importance Research Articles

The importance of quarantine: A bifurcation analysis and modeling of the transmission dynamics of Covid 19

The research aims to construct a mathematical model for COVID 19 that includes features six compartments to evaluate the positive effects of quarantine measures. The model categorizes individuals into thefollowing classes: susceptible, exposed, quarantined, asymptomatic cases, symptomatic cases, and recovered(SEQI1I2R). Several assumptions regarding positivity and boundness are identified to ensure that the solutionoriginated within a certain class and that the basic reproduction number is analyzed. Of course, the existenceof an endemic equilibrium is argued, which provides an understanding of the long-term persistence of thedisease. More precisely, to enhance our understanding of the model’s dynamics, we have analyzed both thelocal and global asymptotic stability of the disease-free equilibrium. Moreover, to assess the global stabilityof the system, we employ a Lyapunov function which provides a comprehensive mathematical evaluation. Atthe same time, our findings show evidence of a backward bifurcation which is recognized as a possible resultof the clinical transition from an asymptomatic state to symptom one.

Differences in pronotum shape in Phlaeothripinae thrips (Thysanoptera) of quarantine importance

Differences in pronotum shape in Phlaeothripinae thrips (Thysanoptera) of quarantine importance

Biology of Citrus long-horned beetle, Anoplophora chinensis (Forster, 1771) (Coleoptera: Cerambycidae) on hazelnut

Anoplophora chinensis (Forster, 1771) (Coleoptera: Cerambycidae) is an important quarantine pest causing significant economic losses. It was detected in hazelnut orchards in Trabzon province in 2017 and an eradication program continues. In this study, the population level and biology of the pest were determined by rearing it and conducting periodic surveys in hazelnut orchards in Trabzon province Maçka district between 2020 and 2021. It has been revealed that the adult emergence of A. chinensis takes place in the last weeks of June and adults are found in nature until the end of August. The oviposition period continued from the end of June to the beginning of September and the larval stage of A. chinensis was observed throughout the year. The pupal stage was observed in May and June. With this study, the biology of A. chinensis in Türkiye has been revealed in detail for the first time and obtained data guides the eradication and surveillance studies.

Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China

Chrysomyxa rusts cause significant damage to spruce in both natural forests and plantations. Particularly, Three Chrysomyxa species, Chrysomyxa deformans, Chrysomyxa qilianensis, and Chrysomyxa rhododendri, listed as National Forest Dangerous Pests in China, have severely affected many economically and ecologically important spruce native species in China. Also, Chrysomyxa arctostaphyli, an important plant quarantine fungus, causes a damaging broom rust disease on spruce. Therefore, rapid, and efficient detection tools are urgently needed for proper rust disease detection and management. In this study, a sensitive, genus-specific loop-mediated isothermal amplification (LAMP) assay targeting the ITS-28S rRNA region was developed to detect the presence of Chrysomyxa in spruce needle and bud samples. After optimization and validation, the LAMP assay was found to be sensitive to detect as low as 5.2 fg/µL DNA, making it suitable for rapid on-site testing for rust infection. The assay was also specific to Chrysomyxa species, with no positive signals from other rust genus/species. The application of LAMP in the early detection of rust infections in spruce needles and buds was investigated, and spatial colonization profiles as well as the means of overwintering of Chrysomyxa woroninii in infected buds and branches were verified using the LAMP assay. This LAMP detection method will facilitate further studies on the characteristics of the life cycle and inoculation of other systemic rusts.

Comparison of different attract-and-kill device densities to control the adult population of Popillia japonica (Coleoptera: Scarabaeidae).

Popillia japonica Newman is a scarab beetle native to Japan that is considered a serious pest outside its native range. It can feed on more than 400 host plants and spread about 10 km per year in invaded territories, therefore it is considered the second most important quarantine pest in Europe. Both chemical and biological insecticides have been used for control, with variable results. Despite ongoing efforts, P. japonica remains a threat in invaded countries, and it is necessary to test more effective and sustainable solutions in the context of integrated pest management. Here we present a study on long-lasting insecticide-treated nets (LLINs) assembled in semiochemical-baited attract-and-kill devices (A&Ks) as a means to control adults of P. japonica with low environmental impact. This study complements previous ones in which we first evaluated the effectiveness of the LLINs in the laboratory and then tested both effectiveness and duration in field-exposed A&Ks against P. japonica. In the present work we compared the effectiveness of three different densities of A&Ks per hectare in areas where the population of P. japonica was numerically homogeneous. The different densities of A&K (1, 2, 4 A&Ks per ha) resulted in an overall reduction of the population of P. japonica by about two thirds in comparison to the control area. This study suggests that the use of one A&K per hectare, requiring minimal management effort, is an effective ratio for reducing local populations of P. japonica. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Integrative identification of Aphelenchoides fragariae (Nematoda: Aphelenchoididae) parasitizing Fuchsia hybrid in China

The strawberry crimp nematode (Aphelenchoides fragariae) is a serious pathogen of ornamental crops and an important quarantine object in approximately 50 countries and regions including China. One nematode population within the genus Aphelenchoides was discovered from diseased leaves of fuchsia plants (Fuchsia × hybrid Voss.) in Chengdu city, Sichuan province of China. Morphological and morphometric data were obtained using light microscopy and scanning electron microscopy. After detailed examination, the species was identified as A. fragariae. Three rDNA sequences of this species, including partial rRNA small subunit, D2-D3 expansion domains of the rRNA large subunit and internal transcribed spacer, were amplified and sequenced. Bayesian trees inferred from these three rDNA sequences were constructed, revealing that this species is placed in a high support monophyletic clade with A. fragariae but clearly separated from all other Aphelenchoides species. Moreover, host-suitability tests showed that the Aphelenchoides population not only can harm and reproduce in F. hybrid, but also in Fragaria ananassa and Pteris vittata (two common hosts of A. fragariae). In conclusion, the study confirmed A. fragariae identity of the nematode from F. hybrid in Chengdu city based on morphology, molecular analysis and host-suitability tests. To our knowledge, this is the first molecular and morphological confirmation of A. fragariae in China, and F. hybrid was first discovered to be attacked by A. fragariae.

Identifying Morphs of the Yellow-Legged Hornet (Vespa velutina) and Other Pests of Quarantine Importance with Geometric Morphometrics

We assess the accuracy of geometric morphometrics (GMM) for determining the origin of insects of quarantine importance using the Asian hornet (Vespa velutina Lep.1836) as a case study. This species is highly variable, has an extensive natural distribution, and has been transported to many regions of the world. Forewing landmarks were applied to a large sample of regionally specific color morphs (previously considered “subspecies”) from across the species’ native Asian range. We reconfirm that GMM can statistically distinguish geographic variants independent of the color patterns that have heretofore been used for provenance, but which have been suspected of being unreliable. Almost all morphs in our analyses were statistically different except the centrally located V. v. variana, whose range lies between the continental V. v. auraria Smith, 1852, and V. v. nigrithorax du Buysson, 1905 morphs, and the Malaysian and Indonesian morphs. Even with moderate-sized training samples, discriminant function analysis (DFA) was able to classify geographic morphos with about 90% accuracy (ranging from 60% to 100%). We apply these results to determine the origin of a dead wasp recently intercepted in a mail parcel in Utah. Both DFA and continuous-trait maximum-likelihood clustering suggest that the Utah specimen belongs to the nigrithorax morph, which is native to southern China but now invasive in Europe, Japan, and Korea. These results are also supported by DNA barcode analysis, which groups the Utah individual with nigrithorax populations in South Korea and Japan. The relationship between variation in wing shape and genetic differentiation deserves further study, but molecular data are consistent with the GMM results suggesting that morphometric comparisons may be able to identify and provenance intercepted specimens quickly and inexpensively when molecular sequences and taxonomic specialists are unavailable.

Morphomolecular characterization of invasive fruit borer infesting apple

Apple, a vital fruit crop worldwide and a major crop grown in northern parts of India, acts as a backbone for the survival and livelihood of the farming community. However, it is prone to severe damage from insect pests and diseases. In the past few years, due to erratic weather patterns, there has been an alarmingly increased infestation of different insect pests, both invasive and non-invasive, resulting in substantial economic losses to this industry. One similar case was seen in the Himalayan region of India, where the apple crop in Karewas was heavily damaged by some invasive fruit borer, feeding on pulp and making galleries to reach the seed locules, thereby destroying the seed material. To ascertain the true identity of this pest, a study based on morpho-molecular characterization of this pest was carried out in the Faculty of Agriculture, Wadura, SKUAST Kashmir, India, during the years 2021 and 2022. The invasive fruit samples were collected from apple orchards at different locations (experimental sites) in North India by installing delta sticky traps @ 5 traps/ha for moth collection. The fruit-boring larvae and pupa were also collected and reared in confined chambers of the laboratory with controlled temperature and humidity. All the laboratory investigations were conducted at the Division of Entomology, FoA, Wadura, SKUAST Kashmir. During the investigation, it was confirmed that the invasive borer is a codling moth (Cydia pomonella L.), a member of the family Tortricidae, order Lepidoptera, which was earlier having quarantine importance in India, as it was only present in the Ladakh region of India. From the phylogenetic analysis of sample sequences, the species of codling moth present at all experimental sites was more identical to the codling moth sequence from Leh (the northernmost arid region), India. Further, the study of life cycle and voltinism revealed that codling moth completes three generations per year in Kashmir, with a single cycle lasting up to 2.5 months. However, the timing of various generations varies, depending on prevailing weather conditions at specific locations and times. The number of generations completed by codling moth in Kashmir conditions during apple growing season was three full and a half overwintering generations. The study of the life cycle of this important pest was necessary to know the weak link for its effective management and to prevent the economic loss in apple fruit.

LAND SNAILS AND SLUGS (MOLLUSCA: GASTROPODA) IN THE INTERNATIONAL LOADING AND UNLOADING AREAS OF PANAMA

This study aims to identify the species of land snails and slugs that may invade Panama through the loading and unloading sites of national and international merchandise in Panama City and Colon, along with their probable places of origin. The investigation also considers the ecological, economic, and health impacts of these potential invasive species. Between September 2008 and September 2009, sampling was conducted at the main seaports and airports of Panama City and Colon, including their containers, warehouses, and surrounding natural habitats. The study utilized GPS for accurate location marking, and sampling occurred twice a week. Specimens were collected, with live material fixed in 70% alcohol, and slugs dissected for identification through sexual organs. Taxonomic identification involved conchological characteristics and expert consultation, with the data analyzed for species abundance correlation with precipitation and humidity. A total of 2379 individuals were collected, identifying seven species across nine genera and four taxonomic families, with the Subulinidae family being the most abundant. Notably, Praticolella griseola, an agricultural pest, and Hapiella w. f. decolorata were new records for Panama. The study found species richness and abundance correlated with rainfall but not relative humidity. The research highlighted the quarantine importance of several species due to their roles as pests and vectors of diseases like Angiostrongylus cantonensis and A. costarricensis. Panama hosts a moderate ecological diversity of gastropods, with certain species posing significant risks to agriculture, public health, and native ecosystems. The study underscores the necessity for ongoing surveillance and management of non-native mollusk introductions, particularly in a global trade hub like Panama. Identifying and understanding the distribution of invasive gastropods are crucial for developing effective control measures and mitigating their impact on the environment and human health.

Consumption of β-Caryophyllene Increases the Mating Success of Bactrocera zonata Males (Diptera: Tephritidae).

The peach fruit fly, Bactrocera zonata (Saunders) (Diptera: Tephritidae), is an economically important polyphagous quarantine pest of horticultural crops endemic to South and Southeast Asia. Methyl eugenol (ME), a naturally occurring phenylpropanoid, is a male attractant used to lure and (when mixed with an insecticide) annihilate the males from the wild population, a method of pest control termed the male annihilation technique (MAT). ME is reported to enhance the mating success of sterile males of Bactrocera spp., which is critical for enhancing the effectiveness of the sterile insect technique (SIT). The suppressed response of ME-treated males to ME-baited traps/devices allows the simultaneous application of the MAT and SIT, increasing the efficiency of area-wide integrated pest management (AW-IPM) programs. However, ME treatment in sterile males in SIT facilities is logistically difficult. β-caryophyllene (BCP) is a widely occurring, safer plant compound and is considered suitable for treating males in SIT facilities. Here, we demonstrate that BCP feeding enhanced B. zonata male mating success to the same extent as ME feeding. Feeding on BCP suppressed the male's subsequent attraction to ME-baited traps, but not to the same degree as feeding on ME. The results are discussed and BCP is suggested as an alternative to ME for the concurrent use of the MAT and SIT.

Types and Fecundity of Neotenic Reproductives Produced in 5-Year-Old Orphaned Colonies of the Drywood Termite, Cryptotermes domesticus (Blattodea: Kalotermitidae)

Orphaned colonies of Cryptotermes domesticus readily produce replacement reproductives and continue propagation. In this study, we aimed to investigate the production and fecundity of neotenic reproductives in 5-year-old colonies of C. domesticus after orphaning. All 15 experimental colonies were successfully re-established by the neotenic reproductive pair. Three types of neotenic reproductives with various wing-bud lengths were observed: type I with micro wing buds, type II with short wing buds, and type III with long wing buds. Four patterns of pairs made up of these neotenics, namely, type I + type II, type I + type III, type II + type II, and type II + type III, exhibited reproductive capacities similar to those of the primary reproductive pair. We speculated that these neotenic reproductives were derived from various nymphal instars. The 5-year-old colonies had three instars of nymphs, with the majority being in the second instar, followed by the first. Thus, the combination of neotenic reproductives with short wing buds and micro wing buds was the dominant differentiation pathway of the orphaned colonies. After the removal of the original primary reproductive pair, the nymphs matured into neotenic reproductives and took over reproduction in the colony in 107.40 ± 15.18 days. This study highlights the importance of quarantine and routine inspection of wood, as well as the significance of early prevention and control of C. domesticus infestation in wood. Moreover, this study confirms the high differentiation and reproductive capacities of C. domesticus.

Quarantine pathogen interceptions on crop germplasm in India during 1986 - 2010 and their possible economic impact

The National Bureau of Plant Genetic Resources Regional Station, Hyderabad has facilitated safe introduction of 356 521 (1986–2010) germplasm accessions of various crops into India. About 60 seed-borne pathogens of quarantine importance were intercepted on crop germplasm during seed examination in the laboratory and post-entry quarantine observations in the field. Of these, the quarantine pathogens, viz. Ralstonia solanacearum and peanut stripe virus in groundnut (Arachis hypogaea L.), Peronospora manshurica (Naum.) Syd. and peanut stripe virus in soybean [Glycine max (L.) Merr.], tobacco streak virus in sunflower (Helianthus annuus L.), Xanthomonas campestris pv holcicola (Elliot) Dye, Burkholderia andropogonis (Smith) Gillis et al. on sorghum [Sorghum bicolor (L.) Moench], Drechslera maydis (Nisik.) Subram. & Jain on maize (Zea mays L.) and Pseudomonas syringae pv tomato (Okabe) Young et al. on tomato (Lycopersicon esculentum Mill.) were intercepted from different countries. Yield losses that could have occurred were estimated had these pathogens of groundnut, soybean, sunflower, maize, sorghum and tomato not been intercepted.

A detection of quarantine scale pest Icerya seychellarum (Hemiptera: Monophlebidae) in South Korea

Abstract Icerya seychellarum (Westwood, 1855) (Hemiptera: Monophlebidae) is found in most tropical and subtropical regions worldwide and is a highly polyphagous pest that affects more than 140 genera of host plants. In 2023, I. seychellarum was discovered for the first time in Seogwipo-si, Jeju Island, South Korea. The specimen was found in heavily infested Cycas revoluta. Here, we provide information regarding the associated host damage, a description of the species, and details of sampling locations. The discovery of I. seychellarum is of agricultural and quarantine importance in South Korea; therefore, this study presents an opportunity to investigate new strategies geared toward advancing efforts to detect and eliminate threats in the future.

Information system related to the phytosanitary condition analysis of territories

The article describes the rationale for the development of an information system in order to perform the tasks of Rosselkhoznadzor on the analysis of phytosanitary status of territories, which includes planning and results of plans for quarantine phytosanitary monitoring of territories, results of phytomonitoring of pests of quarantine importance for major importing countries of Russian grain and analysis of quarantine phytosanitary status of territories. Functions, structure and information objects of this information system, roles of information system users are described. The results of information system implementation in the practice of the Russian Plant Quarantine Service are evaluated, such as increase of efficiency of the Rosselkhoznadzor function on control and supervision in the field of plant quarantine, ensuring compliance with quarantine phytosanitary requirements of importing countries, which significantly increased the export potential of the Russian Federation, significantly reduce the risk of damage caused by the spread of quarantine objects on the territory of the Russian Federation.

Attractant potential of Enterobacter cloacae and its metabolites to Bactrocera dorsalis (Hendel).

Bactrocera dorsalis (Hendel) has a wide host range. It has been the most important quarantine pest in many countries or regions. Currently, chemical control and bait trapping are mainly used in the monitoring, prevention, and control of B. dorsalis. However, chemical control will cause pollution of the environment and drug resistance of insects. Methyl eugenol, the main attractant currently used, can only attract males of B. dorsalis. This study focused on the attractant function and active substances of one key intestinal bacterium, Enterobacter cloacae, which was isolated from B. dorsalis. First, the attraction of the E. cloacae autoclaved supernatant to male and female adults of 0, 6, and15days post-emergence was confirmed using a Y-type olfactometer. Subsequently, through metabolome sequencing and bioassays, L-prolinamide was identified and confirmed as the most effective attractant for B. dorsalis. Finally, the synergistic effect of L-prolinamide with the sex attractant ME was validated through field experiments. This study confirmed the attraction effect of E. cloacae on B. dorsalis and also proved the attraction effect of L-prolinamide, the metabolite of E. cloacae, on B. dorsalis. This laid a theoretical foundation for the development of a new attractant and safe, green, and efficient prevention and control technology of B. dorsalis.

The Effects of Nine Compounds on Aldehyde-Oxidase-Related Genes in Bactrocera dorsalis (Hendel).

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) (B. dorsalis) is an important agricultural, major invasive, and quarantine pest that can cause significant damage to the economic value of the fruit and vegetable industry. Male bait is one of the most effective methods of surveying, monitoring, and controlling B. dorsalis. In our study, we constructed cDNA libraries using total RNA extracted independently from the antennae, mouthparts, and thoracic legs of male and female adults and the ovipositors of female adults and screened out four aldehyde-oxidase-related genes (AOX-related), C58800, C66700, C67485, and C67698. Molecular docking predictions showed that eight compounds, including 3,4-dimethoxycinnamyl alcohol, 3,4-dimethoxy-cinnamaldehyde, deet, ethyl N-acetyl-N-butyl-β-alaninate, n-butyl butyrate, n-butyl butyrate, ethyl butyrate, methyl eugenol, and ethyl acetate, could combine with proteins encoded by the four B. dorsalis AOX-related genes. Furthermore, QPCR was performed to confirm that four compounds, including 3,4-dimethoxy cinnamic aldehyde, butyl levulinic acid ethyl ester (mosquito repellent), butyl butyrate, and methyl eugenol, induced significant changes in the AOX-related genes of B. dorsalis. These results provide useful information and guidance for the batch screening of potentially useful compounds and the search for effective attractants of B. dorsalis.

Virulence of entomopathogenic bacteria Serratia marcescens against the red palm weevil, Rhynchophorus ferrugineus (Olivier).

The red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier), is an important quarantine pest, which has caused serious economic losses in various palm species, such as coconut, oil palm and date palm. Finding effective biocontrol resources is important for the control of this pest and the protection of palm crops. A pathogenic strain HJ-01 was isolated from infected and dead pupa of Tenebrio molitor using tissue separation method. The HJ-01 strain was streak cultured and purified, and its morphological, physiological, biochemical characteristics, and 16S rDNA homology were identified after conducting a pathogenicity test on RPW larvae. Strain HJ-01 exhibited remarkable pathogenicity against RPW larvae. Under the concentration of HJ-01 suspension was 1.0 × 108 cfu/mL, the mortality rate of RPW reached 82.22%, and the half-lethal time (LT50) was 4.72 days. RPW larvae infected with strain HJ-01 showed reduced movement, decreased appetite, and eventual death. As the treatment progresses, the larvae's bodies turned red, became soft, and started to rot, resulting in the discharge of liquid. HJ-01 demonstrated the ability to produce scarlet pigment after 24 h of culture on a basic medium. Colonies of HJ-01 appeared convex, bright red, moist, and viscous, opaque in the center, irregular at the edges, and emitted an unpleasant odor. Under microscopic observation, the cells of HJ-01 appeared as short rod-shaped and flagellate, with a size ranging from (1.2-1.8) μm × (1.0-1.2) μm. Genomic DNA extraction was performed on the strain, and the 16S rDNA sequence was amplified, yielding a sequence length of 1445 bp. The sequence of strain HJ-01 displayed a 99.72% similarity to that of Serratia marcescens. Phylogenetic tree analysis further confirmed that strain HJ-01 belonged to S. marcescens. Therefore, the strain HJ-01 has a very good lethal effect on RPW larvae, and it may be used as an effective bacterium for the control of RPW.

UDA-FlyRecog: Unsupervised domain adaptation for drosophila cross-domain recognition model

Drosophila is an important quarantine pest, and its identification plays a crucial role in agricultural development. To address the issue of significant style variations in Drosophila image samples collected from different environments, which results in poor performance when applying models trained on the source domain to the target domain, this paper proposes a domain-adaptive model for cross-domain Drosophila recognition (UDA-FlyRecog). This model combines global feature alignment and class-aware feature alignment methods, and adopts a cyclic iterative training approach to mitigate the effects of domain shift. Experimental results demonstrate that our method achieves the highest accuracy improvement of up to 16.7% compared to other methods on Drosophila datasets collected from both laboratory and natural environments, indicating promising practical applications.

Prioritisation of quarantine pest list for the Caribbean using a multi-criteria decision approach

Quarantine plant pests are socially, economically and environmentally important due to their impact on food security, human health, global trade and crop production costs. The increase in global trade and tourism, frequent occurrence of natural disasters and climate changes have exacerbated the rate of entry, establishment and spread of plant pests regionally and globally. It has, therefore, become exigent to develop a list of pests of quarantine importance at the regional and national levels to prioritise and allocate the limited available resources to manage the associated risks. In the present study, the Technical Committee on the Formulation and Prioritisation of a Regional Priority Pest List for the Caribbean, in collaboration with the National Plant Protection Organisation of the Caribbean countries and the United States Department of Agriculture - Animal and Plant Health Inspection Service (USDA-APHIS), developed and prioritised a quarantine pest list using a multi-criteria decision-making approach. The technical committee successfully evolved the process in 2014 and 2018 and developed a list of the top 10 pests of quarantine importance for the Caribbean Region, employing the Delphi Technique (DT) and Analytical Hierarchy Process (AHP) through the assignment of criteria that are relevant to the region. The Mediterranean fruit fly (Ceratitis capitata), frosty pod rot (Moniliophthora roreri) and the tomato leaf miner (Tuta absoluta), listed as top quarantine pest threats, were subsequently detected in the region. This exercise guided the authorities in advance to allocate resources and to develop response plans including capacity building for surveillance and detection of priority pests. This has demonstrated the significance and appropriateness of the multi-criteria decision approach to determine priority pest lists and prepare the region for development of better management practices.

Is there hybridization between 2 species of the same genus in sympatry?-The genetic relationships between Anoplophora glabripennis, Anoplophora chinensis, and putative hybrids.

Anoplophora glabripennis (Asian longhorn beetle, ALB) and Anoplophora chinensis (Citrus longhorn beetle, CLB) are native forest pests in China; they have become important international quarantine pests. They are found using the same Salix aureo-pendula host tree of Cixi, Zhejiang province, China. On this host tree, we collected additional beetles that appeared to be morphologically intermediate between ALB and CLB. By using a stereoscope, we observed that there were several bumps on the base of the elytra, which was inconsistent with ALB, which typically has a smooth elytral base, but was more like CLB, which has numerous short tubercles on the elytral base. Given their sympatry and intermediate morphology, we hypothesized that these may represent ALB×CLB hybrids. We studied the genomic profiles for 46 samples (ALB, CLB, and putative hybrids) using genotyping-by-sequencing (GBS) providing a reduced representation of the entire genome. Employing principal component analyses on the 163 GBS-derived single nucleotide polymorphism data, we found putative hybrids tightly clustered with ALB, but genetically distinct from the CLB individuals. Therefore, our initial hybrid hypothesis was not supported by genomic data. Further, while mating experiments between adult ALB and CLB were successful in 4 separate years (2017, 2018, 2020, and 2021), and oviposition behavior was observed, no progeny was produced. Having employed population genomic analysis and biological hybridization experiments, we conclude that the putative hybrids represent newly discovered morphological variants within ALB. Our approach further confirmed the advantage of genome-wide information for Anoplophora species assignment in certain ambiguous classification cases.