Optimal parameters for the extraction of nematodes from wood substrate
https://doi.org/10.31016/1998-8435-2025-19-4-490-499
Abstract
The purpose of the research is determine the optimal time and temperature parameters for the extraction of Bursaphelenchus xylophilus nematodes by Baermann-funnel method from pine wood of Pinus sylvestris L.
Materials and methods. P. sylvestris logs 22–25 cm long, cut from a 15-year-old visually healthy pine tree, were infested with the pine wood stem nematode B. xylophilus and incubated for 45 days in a thermostat at 27 °C. Further, sawdust was obtained by drilling nematode-infested logs with a drill to form a single wood substrate sample. Berman's funnel technique was used to extract nematodes from the wood sample. The experiments included four replications of 8 samples each. Nematode abundance was counted after 6, 24 and 48 hours at 18 and 25 °C. The data were processed in MS Excel.
Results and discussion. The conducted studies showed that about 80% of the nematodes were extracted within the first 24 hours. This period is optimal for nematological analysis. The number of nematodes extracted in the following 24 hours was about 20%. Temperature between 18 and 25 °C influenced the degree of nematode extraction from the wood substrate, but not always significantly. The numbers of extracted nematodes in different samples obtained from the single mixed substrate differed significantly, which can be explained by the aggregation of their distribution in the wood substrate.
About the Authors
A. A. Chalkin
All-Russian Plant Quarantine Center
Россия
Россия
Chalkin Andrey A., Researcher, Forest Quarantine Department
Researcher ID: AET-9683-2022,
Scopus ID:57220116459
Moscow Region
E. N. Arbuzova
All-Russian Plant Quarantine Center
Россия
Россия
Arbuzova Elena N., Candidate of Biological Sciences, Senior Researcher, Head of the Forest Quarantine Department
Researcher ID: I-7153-2015,
Scopus ID: 57222500674
Moscow Region
N. I. Kozyreva
All-Russian Plant Quarantine Center
Россия
Россия
Kozyreva Natalya I., Candidate of Biological Sciences, Researcher, Forest Quarantine Department
Researcher ID: JWP-5077-2024,
Scopus ID: 57190217222
Moscow Region
O. A. Kulinich
All-Russian Plant Quarantine Center;
Center for Parasitology, Institute of Ecology and Evolution. A. N. Severtsov Russian Academy of Sciences
Россия
Россия
Kulinich Oleg A., Doctor of Biological Sciences, Chief Researcher, Forest Quarantine Department
Researcher ID: A-5534-2016,
Scopus ID: 11940984400
Moscow Region; Moscow
References
1. Akhmatovich N. A., Ryss A. Yu. Pathways of spread of stem nematodes of the genus Bursaphelenchus of the xylophilus species group with sawn timber on the territory of the Russian Federation. Parazitologija = Parasitology. 2009; 43 (6): 437–444. (In Russ.).
2. Kiryanov E. S., Krall E. L. Parasitic nematodes of plants and measures to combat them. Leningrad: Nauka, 1969; V. 1. 447 p. (In Russ.).
3. Kudrin A. A., Sushchuk A. A. Methods for studying soil nematode communities. Russian Journal of Ecosystem Ecology. 2022; 7 (2): 44–71. (In Russ.) https://doi.org/10.21685/2500-0578-2022-2-5
4. Kulinich O. A. Methodical guidelines for the detection, determination of parasitic nematodes of forest tree species and methods of protection against them. M., 1990; 70 p. (In Russ.).
5. Kulinich O. A., Arbuzova E. N., Chalkin A. A., Kozyreva N. I., Ryss A. Yu. Distribution of pine stem nematode Bursaphelenchus xylophilus in the world and the results of a survey of coniferous plantations in the Russian Federation. «Sovremennyye problemy lesozashchity i puti ikh resheniya»: materialy II Mezhdunarodnoy nauchnoprakticheskoy konferentsii, posvyashchennoy 95-letiyu so dnya rozhdeniya prof. N. I. Fedorova i 90-letiyu kafedry lesozashchity i drevesinovedeniya = «Modern problems of forest protection and ways to solve them»: Proc. II Int. scientific-practical. conf., dedicated to the 95th anniversary of the birth of prof. N. I. Fedorov and the 90th anniversary of the dep. forest protection and wood science. Minsk: BSTU, 2020; 363–367. (In Russ.).
6. Tabolin S. B., Romanenko N. D., Mityushev I. M. Agronematologiya: Tutorial / General editor S. B. Tabolin. Publ.: ZGAU-MSHA, 2017; 200 p. (In Russ.).
7. Chalkin A. A., Zinnikov D. F., Lyabzina S. N., Sinkevich O. V. Pests and diseases of forest biocenoses of specially protected natural areas of the Republic of Karelia. Fitosanitariya. Karantin rasteniy = Phytosanitary. Plant quarantine. 2021; (2): 9–19. (In Russ.) https://doi.org/10.69536/FKR.2021.30.26.001
8. Chalkin A. A., Kozyreva N. I., Kulinich O. A., Arbuzova E. N. Xylobiont nematodes associated with bark beetles of the genus Ips De Geer, 1775. Fitosanitariya. Karantin rasteniy = Phytosanitary. Plant quarantine. 2024; S4-1 (20): 85. (In Russ.).
9. Shesteperov A. A., Lychagina S. V., Shchitkov G. S. On the epiphytotic process in bursaphelenchiasis (Bursaphelenchus mucronatus) of Scots pine. Rossiyskiy parazitologicheskiy zhurnal = Russian Journal of Parasitology. 2024; 18 (4): 463–474. (In Russ.) https://doi.org/10.31016/1998-8435-2024-18-4-463-474
10. Shesteperov A. A., Savotikov Yu. F. Quarantine phytohelminthoses. Book 1. M.: Kolos, 1995; 463 p. (In Russ.).
11. Arbuzova E. N., Karagyan G. H., Kozyreva N. I., Shchukovskaya A. G., Ghrejyan T. L., Kalashian M. Y., Akopyan K. V. First finding of Bursaphelenchus xylophilus in pine plantations of the Republic of Armenia. Journal of Nematology. 2025; 57 (1): 20250004. https://doi.org/10.2478/jofnem-2025-0004
12. Baermann G. Eine einfache Methode zur Auffindung von Anklostomum (Nematoden) Larven in Erdproben. Tijdschr Diergeneeskd. 1917; 57: 131–137.
13. Cesarz S., Schulz A. E., Beugnon R., Eisenhauer N. Testing soil nematode extraction efficiency using different variations of the Baermann-funnel method. Soil organisms. 2019; 91: 61–72. https://doi.org/10.25674/so91201
14. De la Fuente B., Saura S. Long-term projections of the natural expansion of the pine wood nematode in the Iberian Peninsula. Forests. 2021; 12 (7): 849. https://doi.org/10.3390/f12070849
15. EPPO. Bursaphelenchus xylophilus. EPPO Global Database. https://gd.eppo.int/taxon/BURSXY/categorization. Accessed 28 May 2025
16. EPPO. PM 7/119 (1) Nematode extraction. Bulletin OEPP/EPPO Bulletin. 2013; 43: 471–495. https://doi.org/10.1111/epp.12077
17. Futai K. Pine wilt disease and the decline of pine forests: a global issue. Cambridge Scholars Publishing, 2021; 201.
18. Hallman J., Viaene N. PM 7/119 (1) Nematode extraction. EPPO Bulletin. 2013; 43: 471–495.
19. Kulinich O.A., Kruglic I., Eroshenko A.S., Kolosova N.V. Occurrence and distribution of the nematode Bursaphelenchus mucronatus in the Russian Far East. Russian Journal of Nematology. 1994; 2 (2): 113–120.
20. Mota M., Braasch H., Bravo M. A., Penas A. C., Burgermeister W., Metge K., Sousa E. First report of Bursaphelenchus xylophilus in Portugal and in Europe. Nematology. 1999; 1: 727–734.
21. Robinson, A. F., Heald C. M. Accelerated movement of nematodes from soil in Baermann funnels with temperature gradients. Journal of Nematology. 1989; 21: 370–378.
22. Rodrigues J. M., Sousa E., Abrantes I. Pine wilt disease: historical everview. In Pine wilt disease in Europe: Biological interactions and integrated management (Sousa E.; Vale F., Abrantes I. Eds.) Lisboa. FNAPF-Federação Nacional das Associações de Proprietários Florestais. 2015; 13–32.
23. Schröder T., McNamara D. G., Gaar V. Guidance on sampling to detect pine wood nematode Bursaphelenchus xylophilus in trees, wood and insects. EPPO Bulletin. 2009; 39 (2): 179–188. https://doi.org/10.1111/j.1365-2338.2009.02287.x
24. Sutherland J. R. A brief overview of the pine wood nematode and pine wilt disease in Canada and the United States. In: Pine wilt disease. (Zhao B.G., Futai K., Sutherland J.R., Takeuchi Y. Еds.) Tokyo. Springer, 2008; 13–17. https://doi.org/10.1007/978-4-431-75655-2_3
25. Zamora P., Rodríguez V., Renedo F., Sanz A. V., Domínguez J. C., Pérez-Escolar G., & Martín A. B. First report of Bursaphelenchus xylophilus causing pine wilt disease on Pinus radiata in Spain. Plant Disease. 2015; 99 (10): 1449-1449. https://doi.org/10.1094/PDIS-03-15-0252-PDN
Review
For citations:
Chalkin A.A., Arbuzova E.N., Kozyreva N.I., Kulinich O.A. Optimal parameters for the extraction of nematodes from wood substrate. Russian Journal of Parasitology. 2025;19(4):490-499. (In Russ.) https://doi.org/10.31016/1998-8435-2025-19-4-490-499
Views:
123
JATS XML
Email this article 