{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T22:55:05Z","timestamp":1789167305699,"version":"build-2803163510"},"reference-count":52,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,6,20]],"date-time":"2022-06-20T00:00:00Z","timestamp":1655683200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100013816","name":"a CRC for Developing Northern Australia","doi-asserted-by":"publisher","award":["CRC-P58588"],"award-info":[{"award-number":["CRC-P58588"]}],"id":[{"id":"10.13039\/100013816","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The Kunjin strain of West Nile virus (WNVKUN) is a mosquito-transmitted flavivirus that can infect farmed saltwater crocodiles in Australia and cause skin lesions that devalue the hides of harvested animals. We implemented a surveillance system using honey-baited nucleic acid preservation cards to monitor WNVKUN and another endemic flavivirus pathogen, Murray Valley encephalitis virus (MVEV), on crocodile farms in northern Australia. The traps were set between February 2018 and July 2020 on three crocodile farms in Darwin (Northern Territory) and one in Cairns (North Queensland) at fortnightly intervals with reduced trapping during the winter months. WNVKUN RNA was detected on all three crocodile farms near Darwin, predominantly between March and May of each year. Two of the NT crocodile farms also yielded the detection of MVE viral RNA sporadically spread between April and November in 2018 and 2020. In contrast, no viral RNA was detected on crocodile farms in Cairns during the entire trapping period. The detection of WNVKUN and MVEV transmission by FTATM cards on farms in the Northern Territory generally correlated with the detection of their transmission to sentinel chicken flocks in nearby localities around Darwin as part of a separate public health surveillance program. While no isolates of WNVKUN or MVEV were obtained from mosquitoes collected on Darwin crocodile farms immediately following the FTATM card detections, we did isolate another flavivirus, Kokobera virus (KOKV), from Culex annulirostris mosquitoes. Our studies support the use of the FTATM card system as a sensitive and accurate method to monitor the transmission of WNVKUN and other arboviruses on crocodile farms to enable the timely implementation of mosquito control measures. Our detection of MVEV transmission and isolation of KOKV from mosquitoes also warrants further investigation of their potential role in causing diseases in crocodiles and highlights a \u201cOne Health\u201d issue concerning arbovirus transmission to crocodile farm workers. In this context, the introduction of FTATM cards onto crocodile farms appears to provide an additional surveillance tool to detect arbovirus transmission in the Darwin region, allowing for a more timely intervention of vector control by relevant authorities.<\/jats:p>","DOI":"10.3390\/v14061342","type":"journal-article","created":{"date-parts":[[2022,6,23]],"date-time":"2022-06-23T01:59:47Z","timestamp":1655949587000},"page":"1342","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Nucleic Acid Preservation Card Surveillance Is Effective for Monitoring Arbovirus Transmission on Crocodile Farms and Provides a One Health Benefit to Northern Australia"],"prefix":"10.3390","volume":"14","author":[{"given":"Nina","family":"Kurucz","sequence":"first","affiliation":[{"name":"Medical Entomology, Centre for Disease Control, Public Health Unit, NT Health, Darwin, NT 0811, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9304-1500","authenticated-orcid":false,"given":"Jamie Lee","family":"McMahon","sequence":"additional","affiliation":[{"name":"Public Health Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, QLD 4108, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0120-6305","authenticated-orcid":false,"given":"Allan","family":"Warchot","sequence":"additional","affiliation":[{"name":"Medical Entomology, Centre for Disease Control, Public Health Unit, NT Health, Darwin, NT 0811, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Glen","family":"Hewitson","sequence":"additional","affiliation":[{"name":"Public Health Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, QLD 4108, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean","family":"Barcelon","sequence":"additional","affiliation":[{"name":"Public Health Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, QLD 4108, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederick","family":"Moore","sequence":"additional","affiliation":[{"name":"Public Health Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, QLD 4108, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jasmin","family":"Moran","sequence":"additional","affiliation":[{"name":"Centre for Crocodile Research, Noonamah, NT 0837, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica J.","family":"Harrison","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2004-4073","authenticated-orcid":false,"given":"Agathe M. G.","family":"Colmant","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyran M.","family":"Staunton","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine, James Cook University, Smithfield, QLD 4878, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott A.","family":"Ritchie","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine, James Cook University, Smithfield, QLD 4878, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Townsend","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine, James Cook University, Smithfield, QLD 4878, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dagmar Meyer","family":"Steiger","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine, James Cook University, Smithfield, QLD 4878, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roy A.","family":"Hall","sequence":"additional","affiliation":[{"name":"School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia, QLD 4072, Australia"},{"name":"Australian Infectious Diseases Centre, University of Queensland, St. Lucia, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8895-2414","authenticated-orcid":false,"given":"Sally R.","family":"Isberg","sequence":"additional","affiliation":[{"name":"Centre for Crocodile Research, Noonamah, NT 0837, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonja","family":"Hall-Mendelin","sequence":"additional","affiliation":[{"name":"Public Health Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, QLD 4108, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1111\/avj.12862","article-title":"First evidence of Kunjin strain of West Nile virus associated with saltwater crocodile (Crocodylus porosus) skin lesions","volume":"97","author":"Isberg","year":"2019","journal-title":"Aust. Vet. J."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Habarugira, G., Moran, J., Colmant, A.M., Davis, S.S., O\u2019Brien, C.A., Hall-Mendelin, S., McMahon, J., Hewitson, G., Nair, N., and Barcelon, J. (2020). Mosquito-Independent Transmission of West Nile virus in Farmed Saltwater Crocodiles (Crocodylus porosus). Viruses, 12.","DOI":"10.3390\/v12020198"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"792","DOI":"10.3201\/eid1805.111720","article-title":"Characterization of virulent West Nile virus Kunjin strain, Australia","volume":"18","author":"Frost","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.3201\/eid2208.151719","article-title":"Virulence and evolution of West Nile Virus, Australia, 1960\u20132012","volume":"22","author":"Prow","year":"2016","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"6255","DOI":"10.3390\/ijerph10126255","article-title":"The changing epidemiology of Kunjin virus in Australia","volume":"10","author":"Prow","year":"2013","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1007\/BF01321074","article-title":"Arboviruses causing human disease in the Australasian zoogeographic region","volume":"136","author":"Mackenzie","year":"1994","journal-title":"Arch. Virol."},{"key":"ref_7","first-page":"1","article-title":"Vectors vs humans in Australia- who is on top down under? An update on vector-borne disease and research on vectors in Australia","volume":"23","author":"Russell","year":"1998","journal-title":"J. Vect. Ecol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1016\/S1286-4579(00)01324-1","article-title":"Arboviruses associated with human disease in Australia","volume":"2","author":"Russell","year":"2000","journal-title":"Microbes Infect."},{"key":"ref_9","first-page":"253","article-title":"The ecology and epidemiology of Kunjin virus","volume":"267","author":"Hall","year":"2002","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/0035-9203(68)90094-1","article-title":"Studies of the epidemiology of arthropod-borne virus infections at Mitchel River Mission, Cape York Peninsula, North Queensland. I. Mosquito collections, 1963\u20131966","volume":"62","author":"Standfast","year":"1968","journal-title":"Trans. R. Soc. Trop Med. Hyg."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1038\/icb.1963.2","article-title":"Studies of arthropod-borne virus infections in Queensland III. Isolation and characterisation of virus strains from wild-caught mosquitoes in North Queensland","volume":"41","author":"Doherty","year":"1963","journal-title":"Austral. J. Exp. Biol."},{"key":"ref_12","first-page":"141","article-title":"Arboviruses and their vectors in Australia: An update on the ecology and epidemiology of some mosquito-borne arboviruses","volume":"83","author":"Russell","year":"1995","journal-title":"Rev. Med. Vet. Entomol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.tmaid.2010.05.005","article-title":"The viruses of Australia and the risk to tourists","volume":"9","author":"Smith","year":"2011","journal-title":"Travel Med. Infect. Dis."},{"key":"ref_14","unstructured":"Hall, R., and Aaskov, J. (2017). Murray Valley encephalitis virus. Feigin and Cherry\u2019s Textbook of Pediatric Infectious Diseases, Elsevier. [8th ed.]."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"90","DOI":"10.5694\/j.1326-5377.1986.tb101087.x","article-title":"Illness caused by Kokobera-like virus in aouth-eastern Australia","volume":"145","author":"Boughton","year":"1986","journal-title":"Med. J. Aust."},{"key":"ref_16","unstructured":"Kurucz, N. Mosquito Investigation and Management Recommendations. Lagoon Crocodile Farm, March 2019. Personal Communication."},{"key":"ref_17","unstructured":"Kurucz, N. Mosquito Investigation and Management Recommendations. Janamba Crocodile Farm, March and April. Personal Communication."},{"key":"ref_18","unstructured":"Kurucz, N. Mosquito Investigation. Darwin Crocodile Farm, April and May. Personal Communication."},{"key":"ref_19","first-page":"1167","article-title":"Determination of mosquito (Diptera: Culicidae) bloodmeal sources in Western Australia: Implications for arbovirus transmission","volume":"46","author":"Johansen","year":"2009","journal-title":"J. Med. Ecol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"844","DOI":"10.4269\/ajtmh.1982.31.844","article-title":"Vector competence of Culex pipiens quinquefasciatus for Murray Valley encephalitis, Kunjin and Ross River viruses from Australia","volume":"31","author":"Kay","year":"1982","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Webb, C., Doggett, S., and Russell, R. (2016). A Guide to Mosquitoes of Australia, CSIRO.","DOI":"10.1071\/9780643104464"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"137","DOI":"10.2987\/8756-971X-28.4s.137","article-title":"The contribution of Culex pipiens complex mosquitoes to transmission and persistance of West Nile virus in North America","volume":"28","author":"Andrealis","year":"2012","journal-title":"J. Am. Mosqu. Control Assoc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1093\/jmedent\/47.4.625","article-title":"Detection of West nile virus RNA in mosquitoes and identification of mosquito blood meals collected at Alligator Farms in Louisiana","volume":"47","author":"Unlu","year":"2010","journal-title":"J. Med. Entomol."},{"key":"ref_24","first-page":"76","article-title":"Arbovirus and vector surveillance in Western Australia, 2004\/05 to 2007\/08","volume":"10","author":"Johansen","year":"2009","journal-title":"Arbovirus Res. Aust."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1111\/j.1440-6055.2009.00711.x","article-title":"Vector competence of Australian Culex gelidus Theobald (Diptera: Culicidae) for endemic and exotic arboviruses","volume":"48","author":"Johnson","year":"2009","journal-title":"Aust. J. Entomol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"333","DOI":"10.4103\/0972-9062.147897","article-title":"Replication potential and different modes of transmission of West nile virus in an Indian strain of Culex gelidus Theobald (Diptera: Culicidae) mosquitoes","volume":"51","author":"Sudeep","year":"2014","journal-title":"J. Vector Borne Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"11255","DOI":"10.1073\/pnas.1002040107","article-title":"Exploiting mosquito sugar feeding to detect mosquito-borne pathogens","volume":"107","author":"Ritchie","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Floridis, J., McGuinness, S., Kurucz, N., Burrow, J., Baird, R., and Francis, J. (2018). Murray Valley encephalitis virus: An ongoing cause of enceohalitis in Australia\u2019s north. Trop. Med. Infect. Dis., 3.","DOI":"10.3390\/tropicalmed3020049"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1186\/s13071-015-1114-9","article-title":"Development and field evaluation of the sentiel mosquito arbovirus capture kit (SMACK)","volume":"8","author":"Johnson","year":"2015","journal-title":"Parasites Vectors"},{"key":"ref_30","first-page":"14","article-title":"Murray Valley encephalitis virus detection using honeybait cards in the Northern Territory in 2013","volume":"1","author":"Kurucz","year":"2014","journal-title":"Mosq. Bites"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1603\/ME12112","article-title":"A simple non-powered passive trap for the collection of mosquitoes for arbovirus surveillance","volume":"50","author":"Ritchie","year":"2013","journal-title":"J. Med. Entomol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1089\/vbz.2013.1373","article-title":"Applications of a sugar-based surveillance system to track arboviruses in wild mosquito populations","volume":"14","author":"Townsend","year":"2014","journal-title":"Vector-Borne Zoonotic Dis."},{"key":"ref_33","unstructured":"Government, N.T. (2015). Medical Entomology Annual Report 2013\/14, Department of Health."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.jviromet.2004.01.007","article-title":"Detection of Australasian Flavivirus encephalitic viruses using rapid fluorogenic TaqMan RT-PCR assays","volume":"117","author":"Pyke","year":"2004","journal-title":"J. Virol. Methods"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.jviromet.2006.03.007","article-title":"A simple method for preparing synthetic controls for conventional and real-time PCR for the identification of endemic and exotic disease agents","volume":"135","author":"Smith","year":"2006","journal-title":"J. Virol. Methods"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.33321\/cdi.2019.43.14","article-title":"Arboviral Diseases and Malaria in Australia, 2014\u20132015: Annual Report of the National Arbovirus and Malaria Advisory Committee","volume":"43","author":"Knope","year":"2019","journal-title":"Commun. Dis. Intell."},{"key":"ref_37","unstructured":"Resources, Northern Territory Department of Primary Industry (2017). Berrimah Virology Laboratory Virology Test. Methods Manual: Virus Neutralisation Tests, Issue 10."},{"key":"ref_38","unstructured":"Lee, D., Hicks, M., Griffiths, M., and Debenham, M. (1989). The Culicidae of the Australasian Region, University of Queensland and University of Sydney in collaboration with Commonwealth Department of Community Services and Health."},{"key":"ref_39","unstructured":"Lee, D., Hicks, M., Griffiths, M., Debenham, M., Bryan, J., and Marks, E. (1988). The Culicidae of the Australasian Region, Commonwealth Department of Health and School of Public Health and Tropical Medicine."},{"key":"ref_40","unstructured":"Lee, D., Hicks, M., Griffiths, M., Debenham, M., Bryan, J., Russell, R., Geary, M., and Marks, E. (1987). The Culicidae of the Australasian Region, Commonwealth Department of Health and School of Public Health and Tropical Medicine."},{"key":"ref_41","unstructured":"Lee, D., Hicks, M., Griffiths, M., Debenham, M., Bryan, J., Russell, R., Geary, M., and Marks, E. (1988). The Culicidae of the Australasian Region, Commonwealth Department of Health and School of Public Health and Tropical Medicine."},{"key":"ref_42","unstructured":"Lee, D., Hicks, M., Griffiths, M., Russell, R., and Marks, E. (1982). The Culicidae of the Australasian Region, Commonwealth Department of Health and Commonwealth Institute of Health, Incorporating School of Public Health and Tropical Medicine."},{"key":"ref_43","unstructured":"Lee, D., Hicks, M., Griffiths, M., Russell, R., and Marks, E. (1984). The Culicidae of the Australasian Region, Commonwealth Department of Health and Commonwealth Institute of Health, Incorporating School of Public Health and Tropical Medicine."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1080\/00313029800169456","article-title":"Identification of Australian arboviruses in inoculated cell cultures using monoclonal antibodies in ELISA","volume":"30","author":"Broom","year":"1998","journal-title":"Pathology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3059","DOI":"10.1093\/nar\/gkf436","article-title":"MAFFT: A novel method for rapid multiple sequence alignment based on fast Fourier transform","volume":"30","author":"Katoh","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1093\/molbev\/mst010","article-title":"MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability","volume":"30","author":"Katoh","year":"2012","journal-title":"Mol. Biol. Evol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1111\/jvec.12222","article-title":"Determining Culex annulirostris larval densities and control efforts across a coastal wetland, Northern Territory, Australia","volume":"41","author":"Kurucz","year":"2016","journal-title":"J. Vect. Ecol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1128\/CMR.18.4.608-637.2005","article-title":"Biological transmission of arboviruses: Reexamination of and new insights into components, mechanisms, and unique traits as well as their evolutionary trends","volume":"18","author":"Kuno","year":"2005","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1186\/s13071-019-3798-8","article-title":"Evaluation of honey-baited FTA cards in combination with different mosquito traps in an area of low arbovirus prevalence","volume":"12","author":"Wipf","year":"2019","journal-title":"Parasites Vectors"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Birnberg, L., Temmam, S., Aranda, C., Correa-Fiz, F., Talavera, S., Bigot, T., Eloit, M., and Busquets, N. (2020). Viromics on Honey-Baited FTA cards as a New Tool for the Detection of circulating Viruses in Mosquitoes. Viruses, 12.","DOI":"10.3390\/v12030274"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1111\/jvec.12343","article-title":"Arbovirus surveillance using FTA cards in modified CO2-baited encephalitis virus traps in the Northern Territory, Australia","volume":"44","author":"Kurucz","year":"2019","journal-title":"J. Vec. Ecol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1186\/s13071-021-04951-8","article-title":"Rapid assessment of West Nile virus circulation in a German zoo based on honey-baited FTA cards in combination with box gravid traps","volume":"14","author":"Fynmore","year":"2021","journal-title":"Parasites Vectors"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/6\/1342\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:35:40Z","timestamp":1760139340000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/6\/1342"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,20]]},"references-count":52,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,6]]}},"alternative-id":["v14061342"],"URL":"https:\/\/doi.org\/10.3390\/v14061342","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,20]]}}}