Figure 1.
Figure 2.
Figure 3.
Figure 4.
Primers and PCR conditions used in this study_
| Target | Primers | Sequence (5′-3′) | PCR conditions | Reference | |||
|---|---|---|---|---|---|---|---|
| Denaturation | Annealing | Extension | Cycles | ||||
| 18S rDNA | MF | CAAGGACGAAAGTTAGAGGTTC | 95°C, 30sec | 50°C, 30sec | 72°C, 45sec | 40 | Kobylinski et al. 2012 |
| MR | GGAAACCTTGTTACGACTTTTA | ||||||
| KuboF | ATGCATGTCTAAGCACATGCC | 95°C, 30sec | 50°C, 1min | 72°C, 2min | 40 | Kubo et al. 2016 | |
| KuboR | AACCTTGTTACGACTTTTACTTC | ||||||
| newF | AGTTATCTACTTGGATAACTGTG | This study | |||||
| 28S rDNA | F (3745-3764) | AGCGGAGGAAAAGAAACTAA | 95°C, 30sec | 62°C, 30sec | 72°C, 1min | 35 | Nadler et al. 1998 |
| R (4359-4377) | ATCCGTGTTTCAAGACGGG | ||||||
| LSU-F | ACAAGTACCGTGAGGGAAAGTTG | 94°C, 30sec | 45°C, 1min | 72°C, 2min | 40 | Tong et al. 2021 | |
| LSU-R | TCGGAAGGAACCAGCTACTA | ||||||
Mermithid nematodes reported in Japan_
| Species | Location | Host | Reference |
|---|---|---|---|
| Agamermis unka | Hiroshima | Sogatella furcifera Nilaparvata lugens (Homoptera: Delphacidae) | Hidaka and Andow, 2017 |
| Amphimermis zuimushi | Shizuoka | Chilo simplex Butler (Lepidoptera: Pyralidae) | Kaburaki and Imamura, 1932 |
| Gastromermis mesostoma lsomermis bipapillatus | Fukuoka, Kagoshima, Oita | Simulium japonicum (Diptera: Simuliidae) | Poinar et al., 1986 |
| Hexamermis sp. Mermithidae sp. | Mie, Saga, Shizuoka | Glaucias subpunctatus Plautia stali (Hemiptera: Parastrachiidae, Pentatomidae) | Watanabe et al., 2021 |
| Mermithidae sp. | Hokkaido | Bombus pseudobaicalensis Vogt (Hymenoptera: Bombinae) | Kubo et al., 2016 |
| Mermithidae sp. | Kanagawa | Ligidium sp. (Isopoda: Ligiidae) | Yoshino and Waki, 2021 |
| Mermithidae sp. | Hokkaido | Eriosoma auratum (Hemiptera: Aphididae) | Tong et al., 2021 |
| Mermithidae sp. | Saga, Nagasaki, Kagoshima | Parastrachia japonensis Plautia stali (Hemiptera: Parastrachiidae, Pentatomidae) | Iryu et al., 2020 |
| Mermithidae sp. | Kyoto | Pardosa pseudoannulata (Araneae: Lycosidae) | Iida et al., 2003 |
Mermithid nematodes used in phylogenetic analysis of 18S and 28S rDNA retrieved from GenBanka_
| Organism | Location | Host | Accession number | Reference | |
|---|---|---|---|---|---|
| 18S rDNA | 28S rDNA | ||||
| Agamermis changshaensis | - | - | DQ628908 | - | Unpublished |
| Agamermis changshaensis | - | - | - | EF617371 | Unpublished |
| Agamermis xianyangensis | - | - | EF617352 | EF617370 | Unpublished |
| Agamermis sp. | Argentina United States | Armadillidium vulgare (Isopoda: Armadillidiidae) | OP380610 | - | Unpublished |
| Megacopta cribraria (Hemiptera: Plataspidae) | KX173336 | - | Stubbins et al. 2016 | ||
| - | Environment | DQ665653 | - | Unpublished | |
| Allomermis solenopsii | - | - | DQ533953 | - | Unpublished |
| Amphimermis enzoni | Argentina | Ischnura fluviatilis Rhionaeschna bonaerensis (Odnata) | MT021436 | - | Rusconi et al. 2020 |
| Amphimermis sp. | China | - | EF617354 | EF617372 | Unpublished |
| - | EF617355 | EF617373 | |||
| Gastromermis sp. | - | - | DQ533954 | - | Unpublished |
| - | - | AY146543 | - | Unpublished | |
| Gastromermis viridis | Canada | Simulium squamosum (Diptera: Simuliidae) | EU792502 | - | St-Onge et al. 2008 |
| Heleidomermis sp. | - | - | DQ533955 | - | Unpublished |
| Hexamermis agrotis | - | - | DQ530350 | - | Unpublished |
| - | - | - | EF617369 | Unpublished | |
| Turkey | - | - | KC784667 | Unpublished | |
| Hexamermis popilliae | Italy | Popillia japonica (Coleoptera: Scarabaeidae) | MF040823 | MF040824 | Mazza et al. 2017 |
| - | MF040826 | ||||
| Hexamermis sp. | Japan | Glaucias subpunctatus (Hemiptera: Pentatomidae) | LC661690 | - | Watanabe et al. 2021 |
| LC661691 | - | ||||
| Isomermis lairdi | Ghana | Simulium squamosum (Diptera: Simuliidae) | FN400899 | - | Crainey et al. 2009 |
| FN400892 | - | ||||
| Limnomermis sp. | - | - | KJ636371 | - | Unpublished |
| Mermis nigrescens | Viet Nam | Orthomorpha coarctata (Polydesmida: Paradoxosomatidae) | OQ341432 | - | Unpublished |
| New | Forficula auricularia (Dermaptera: Forficulidae) | KF583882 | KF886019 | Unpublished | |
| Zealand | Plant | KF583883 | KF886018 | ||
| - | - | AF036641 | - | Blaxter et al. 1998 | |
| Mermithidae sp. | Japan | Parastrachia japonensis (Hemiptera: Parastrachiidae) | LC512368 | - | Iryu et al. 2020 |
| LC512369 | - | ||||
| LC512370 | - | ||||
| LC512371 | - | ||||
| Japan | Plautia stali (Hemiptera: Parastrachiidae) | LC512372 | - | Iryu et al. 2020 | |
| Japan | Ligidium sp. (Isopoda: Ligiidae) | LC596451 | LC500234 | Yoshino and Waki, 2021 | |
| LC596452 | LC500235 | ||||
| Japan | Eriosoma auratum (Hemiptera: Aphididae) | MW649131 | MW653323 | Tong et al. 2021 | |
| Japan | Bombus pseudobaicalensis (Hymenoptera: Apidae) | LC114020 | - | Kubo et al. 2016 | |
| Japan | Glaucias subpunctatus (Hemiptera: Pentatomidae) | LC661692 | - | Watanabe et al. 2021 | |
| Japan | Plautia stali (Hemiptera: Parastrachiidae) | LC661693 | - | Watanabe et al. 2021 | |
| Japan | Stomoxys calcitrans (Diptera: Muscidae) | LC788412 | LC788415 | This study | |
| LC788413 | LC788416 | ||||
| LC788414 | LC788417 | ||||
| Australia | Kosciscola tristis (Orthoptera: Acrididae) | JQ894731 | - | Umbers et al. 2015 | |
| JQ894732 | - | ||||
| Mesomermis camdenensis | Canada | Simulium squamosum (Diptera: Simuliidae) | EU792504 | - | St-Onge et al. 2008 |
| Mesomermis flumenalis | Canada | Simulium squamosum (Diptera: Simuliidae) | EU792506 | - | St-Onge et al. 2008 |
| Octomyomermis huazhongensis | - | - | EF617353 | EF617368 | Unpublished |
| Ovomermis sinensis | China | Spodoptera frugiperda (Lepidoptera: Noctuidae) | MN367957 | MN367955 | Sun et al. 2020 |
| MN367956 | MN367954 | ||||
| Poland | Cerapteryx graminis (Lepidoptera: Noctuidae) | KU177046 | - | Unpublished | |
| - | - | DQ520879 | - | Unpublished | |
| Pheromermis sp. | France | Vespa velutina (Hymenoptera: Vespidae) | KR029620 | KR029622 | Villemant et al. 2015 |
| KR029621 | KR029623 | ||||
| Romanomermis culicivorax | - | Culex sp. (Diptera: Culicidae) | DQ418791 | - | Unpublished |
| - | - | - | EF417153 | Sonnenberg et al. 2007 | |
| Romanomermis iyengari | - | - | JX021620 | - | Unpublished |
| Romanomermis sichuanensis | - | Culex sp. (Diptera: Culicidae) | EF612769 | EF617366 | Unpublished |
| Romanomermis wuchangensis | - | Culex quinquefasciatus (Diptera: Culicidae) | DQ520878 | EF617365 | Unpublished |
| Strelkovimermis spiculatus | Argentina | Aedes albifasciatus (Diptera: Culicidae) | KP270703 | - | Belaich et al. 2015 |
| - | Aedes sp. (Diptera: Culicidae) | DQ665654 | - | Unpublished | |
| Thaumamermis cosgrovei | - | - | DQ665655 | - | Unpublished |
| Thaumamermis zealandica | New Zealand | Bellorchestia quoyana (Amphipoda: Talitridae) | KY264164 | KY264165 | Unpublished |
Detection of parasitic nematodes from Stomoxys calcitrans on three farms in the Gifu Prefecture, Japan_
| Farm | Location | Sampling date | Total stable flies (Number) | Parasitized stable flies (Number) | Total nematodes (Number) |
|---|---|---|---|---|---|
| Dairy cattle farm | |||||
| F1 | Gifu | 11 Oct 2021 | 80 | 1 | 1a |
| 15 Sep 2022 | 140 | 0 | 0 | ||
| 20 Sep 2023 | 112 | 0 | 0 | ||
| Subtotal | 332 | 1 | 1 | ||
| F2 | Yourou | 8 Oct 2021 | 17 | 1 | 1b |
| 18 Oct 2022 | 38 | 0 | 0 | ||
| 11 Oct 2023 | 27 | 0 | 0 | ||
| Subtotal | 82 | 1 | 1 | ||
| Sheep farm | |||||
| F3 | Nakatsugawa | 29 Jul 2022 | 72 | 1 | 1c |
| Total | 486 | 3 | 3 | ||
Information on locations of nematode species and hosts corresponding to nematodes used in Figure 4a_
| Species | Accession number | Location in Japan | Host | Reference | ||
|---|---|---|---|---|---|---|
| (Country) | Family | Genus | Species | |||
| Ovomermis / Hexamermis cluster | ||||||
| Ovomermis sinensis | KU177046 | (Poland) | Noctuidae | Cerapteryx | C. graminis | Unpublished |
| Hexamermis sp. | LC661690 | Mie | Pentatomidae | Glaucias | G. subpunctatus | Watanabe et al., 2021 |
| LC661691 | Mie | Pentatomidae | Glaucias | G. subpunctatus | Watanabe et al., 2021 | |
| Mermithidae sp. | LC788413 | Gifu | Muscidae | Stomoxys | S. calcitrans | This study |
| LC788412 | Gifu | Muscidae | Stomoxys | S. calcitrans | This study | |
| LC512368 | Saga | Parastrachiidae | Parastrachia | P. japonensis | Iryu et al., 2020 | |
| LC512372 | Saga | Parastrachiidae | Parastrachia | P. japonensis | Iryu et al., 2020 | |
| LC661692 | Shizuoka | Pentatomidae | Glaucias | G. subpunctatus | Watanabe et al., 2021 | |
| LC114020 | Hokkaido | Apidae | Bombus | B. pesudobaicalensis | Kubo et al., 2016 | |
| LC661693 | Mie | Pentatomidae | Plautia | P. stali | Watanabe et al., 2021 | |
| LC596451 | Kanagawa | Ligiidae | Ligidium | sp. | Yoshino and Waki, 2021 | |
| LC596452 | Kanagawa | Ligiidae | Ligidium | sp. | Yoshino and Waki, 2021 | |
| Hexamermis agrotis | DQ530350 | (China) | - | - | - | Unpublished |
| Hexamermis popilliae | MF040823 | (Italy) | Scarabaeidae | Popillia | P. japonica | Mazza et al., 2017 |
| Amphimermis cluster | ||||||
| Amphimermis enzoni | MT021436 | (Argentina) | Coenagrionidae | Ischnura | I. fluviatilis | Rusconi et al., 2020 |
| Aeshnidae | Rhionaeschna | R. bonariensis | ||||
| Mermithidae sp. | LC788414 | Gifu | Muscidae | Stomoxys | S. calcitrans | This study |
| LC512369 | Saga | Parastrachiidae | Parastrachia | P. japonensis | Iryu et al., 2020 | |
| LC512371 | Kagoshima | Parastrachiidae | Parastrachia | P. japonensis | Iryu et al., 2020 | |
| LC512370 | Kagoshima | Parastrachiidae | Parastrachia | P. japonensis | Iryu et al., 2020 | |
| Others | ||||||
| Mermithidae sp. | MW649131 | Hokkaido | Aphididae | Eriosoma | E. auratum | Tong et al., 2021 |
| Romanomermis sichuanensis | EF612769 | (China) | - | - | - | Unpublished |