BACKGROUND: Khapra beetle (Trogoderma granarium Everts) is a significant pest of food products around the world, causing great losses of stored grain and produce, with export restrictions imposed on countries with established beetle populations. Khapra beetle is a high-priority exotic invertebrate pest in many countries requiring a rapid quarantine/biosecurity response when incursions occur. To address this, we developed a novel Khapra LAMP (loop-mediated isothermal amplification) assay using a portable real-time fluorometer and an additional 18S ribosomal DNA (18S) insect control LAMP assay for confirmation of the presence of insect DNA. Both LAMP tests can be performed either in a portable real-time fluorometer or using simple, visual colorimetric technique.RESULTS: Both the Khapra and 18S LAMP tests amplify positive samples within ≤ 25 min, with an anneal derivative temperature of 77.7 ± 0.7 °C for Khapra LAMP test and 88.0 ± 1.0 °C for 18S. The new Khapra LAMP assay is sensitive to very low levels of DNA (1.02 × 10 −6 ng ∼L −1 ). Additionally, we developed a gBlock double stranded DNA fragment for use as positive Khapra control with a different anneal derivative of 80 °C. Both assays are simple to use in the field and are capable of amplifying DNA from target beetles, even when samples are partially degraded which is typically found during surveillance activities. By screening a broad panel of Dermestidae species we demonstrate that our new assay is species-specific, with no detections of false positives. Also, we evaluated multiple DNA extraction methods, with both QuickExtract and HotSHOT extraction methods proving suitable for in-field use. CONCLUSION:The novel Khapra and 18S LAMP assays should improve speed, accuracy and confidence of detection of Khapra beetle at incursion points and aid rapid biosecurity responses in any country affected, especially as the assays described here are portable and easy to implement in the field conditions where resources are limited.

Male sciarids collected in Australia from inside post-entry quarantine and domestic greenhouses and from vegetable gardens and various plants, were slide mounted and identified. Specimens intercepted during on-arrival biosecurity inspections of imported nursery stock plants were also examined, and the New South Wales Department of Primary Industries collection of slide-mounted Sciaridae was reviewed. Plant and mushroom pest species that are present in Australia are Bradysia impatiens (Johannsen), B. ocellaris (Comstock), Lycoriella agraria (Felt), L. ingenua (Dufour) = Sciara womersleyi Séguy, 1940 syn. n., L. sativae (Johannsen) = Sciara auberti Séguy, 1940 syn. n., Sciara jeanneli Séguy, 1940 syn. n., Sciara solispina Hardy, 1956 syn. n., and Cosmosciara hartii (Johannsen, 1912) comb. n. = Plastosciara perniciosa Edwards, 1922 syn. n. The last species is a new record for Australia. Bradysia tilicola (Loew) and Pnyxia scabiei (Hopkins) are potential pest species, but they have not been reported yet from Australia. An identification key to enable separation of the pest species is provided. Species with uncertain connections to plant and mushroom cultures are B. pallipes (Fabricius), B. strenua (Winnertz, 1867) = B. watsoni Colless, 1962 syn. n., Corynoptera concinna (Winnertz), (all three species are new records for the Australian mainland) and Hyperlasion aliens Mohrig (a new record for Tasmania). Bradysia spatitergum (Hardy) and Scatopsciara atomaria (Zetterstedt) were intercepted during the on-arrival biosecurity inspections of live plants imported from China and Canada respectively. Both species are widespread overseas but are not known to occur in Australia.

A total of 27 male sciarid types described by Skuse (1888 and 1890), held in the Australian National Insect Collection, Canberra, and the Australian Museum, Sydney, were remounted and examined microscopically. Of these, 25 species were described as Sciara Meigen, one as Zygoneura Meigen and one as Trichosia Winnertz. Revision of these species revealed the following: 13 species belong to the genus Bradysia Winnertz (B. amabilis, B. conjuncta, B. crassicornis, B. exsequialis, B. frequens, B. froggatti, B. luctifica, B. maesta, B. mastersi, B. ornatula, B. pernitida, B. pictipes, B. unica), 1 species to the genus Corynoptera Winnertz (C. minutela), 4 species to the genus Austrosciara Schmitz & Mjöberg (Aus. infrequens, Aus. montivaga, Aus. spectabilis, Aus. winnertzi), 2 species to the genus Pseudolycoriella Menzel & Mohrig (Psl. cavatica, Psl. ignobilis), 1 species to the genus Pseudozygomma Mohrig (Pseudoz. maculipennis), 1 species to the genus Sciara Meigen (Sc. tryoni), and 1 species to the genus Scythropochroa Enderlein (Scyth. macleayi). In total 26 species were new combinations. Eight species names were declared as new synonyms: Bradysia pictipes (Skuse, 1888) = Sciara notata Skuse, 1888 syn. n. and = Bradysia seticornis Vilkamaa, Hippa & Mohrig, 2012 (from New Caledonia) syn. n.; Bradysia conjuncta (Skuse, 1890) = Sciara serenipennis Skuse, 1890 syn. n.; Pseudolycoriella cavatica (Skuse, 1888) = Sciara familiaris Skuse, 1888 syn. n. and = Sciara festiva Skuse, 1888 syn. n.; Bradysia luctifica (Skuse, 1888) = Bradysia planistylata Vilkamaa, Hippa & Mohrig, 2012 syn. n.; Sciara tryoni Skuse, 1890 = Sciara insulana Vilkamaa, Hippa & Mohrig, 2015 syn. n. (both species are from New Caledonia); Austrosciara winnertzi (Skuse, 1888) = Sciara rufulenta Edwards, 1927 syn. n. (from New Zealand). Lectotype specimens were designated for 17 species in order to fix the names.

The genus Pseudolycoriella Menzel & Mohrig, 1998 is distributed worldwide and especially species rich within the southern hemisphere. Within the Australasian region it is known from New Zealand (Mohrig & Jaschhof 1999; 5 species), New Caledonia (Köhler & Menzel 2013; 14 species) and Papua-New Guinea (Mohrig 2013; 22 species). Here we report the first records of the genus from Norfolk Island and Australia.

On the Hawaiian Islands 22 sciarid species were detected, belonging to the following ten genera: Austrosciara Schmitz & Mjöberg, Bradysia Winnertz, Corynoptera Winnertz, Cratyna Winnertz, Epidapus Haliday, Hyperlasion Schmitz, Lycoriella Frey, Phytosciara Frey, Pseudolycoriella Menzel & Mohrig and Scatopsciara Edwards. The revision resulted in new combinations for the following five species: Austrosciara hawaiiensis (Hardy) comb. n., Corynoptera prominens (Hardy) comb. n., Cratyna adrostylata (Hardy) comb. n., Cr. longicosta (Hardy) comb. n., and Scatopsciara hoyti (Hardy) comb. n. Eight species were declared as new synonyms: Bradysia bishopi Steffan, 1973 = B. centidens Vilkamaa, Hippa & Mohrig, 2012 syn. n.; B. crassicornis (Skuse, 1890) = B. molokaiensis (Grimshaw, 1901) syn. n. and = B. aspercera Mohrig, 2016 syn. n.; B. radicum (Brunetti, 1912) = B. spatitergum (Hardy, 1956) syn. n.; Corynoptera prominens (Hardy, 1956) = C. gladiota Mohrig, 2004 syn. n.; Cosmosciara hartii (Johannsen, 1912) = Plastosciara (Plastosciara) latipons Hardy, 1956 syn. n.; Hyperlasion wasmanni (Schmitz, 1918) = Scythropochroa magnisensoria Hardy, 1956 syn. n.; and Scatopsciara hoyti (Hardy, 1956) = Sc. spiculata Vilkamaa, Hippa & Mohrig, 2012 syn. n. These four species are new reports for Hawai‘i, three of them are new to science: Epidapus pallidus (Séguy), Pseudolycoriella nigrofemoralis Mohrig, Kauschke & Broadley sp. n., Scatopsciara hardyi Mohrig, Kauschke & Broadley sp. n. and Sc. steffani Mohrig, Kauschke & Broadley sp. n. A lectotype was designated for Bradysia radicum (Brunetti) in order to fix the name. All new and revised species are figured.        The species Bradysia bishopi Steffan, 1973, B. ocellaris (Comstock, 1882), B. radicum (Brunetti, 1912), Cosmosciara hartii (Johannsen, 1912), Pseudolycoriella planiforceps (Steffan, 1971) and Scatopsciara steffani Mohrig, Kauschke & Broadley sp. n. are reported from the Galápagos Islands.