The prevalence of plant parasitic nematodes (PPN) in the Irish peatlands was investigated in five different peatland habitats—raised bog, cutover scrub/woodlands, fens and peat grasslands, which were further sub-categorised into fourteen different sub-habitats. Within the raised bog habitat were healthy bog hummock (HBH), healthy bog lawn (HBL), degraded bog hummock (DBH) and degraded bog lawn (DBL) and the fen habitats were fen peat (FP) and rich fen peat (R-FP). Cutover scrub or woodland habitat included cutover scrub rewetted (C-RW), cutover scrub non-rewetted (C-NRW), woodlands rewetted (W-RW) and woodlands non-rewetted (W-NRW). Grassland included wasted peat (WP), rough grazing (RG-I) and improved fen peat grassland (IFPG-RW and IFPG-NRW). Soil samples from peatlands were all collected between July and December 2023 when the temperature ranged from 12 to 20 °C. One half of each sample was used for molecular nematode analysis and the other half for morphological identification of nematodes. For the morphological identification, a specific nematode extraction protocol was optimised for peatland soils, and the extracted nematodes were fixed onto slides to be studied under a high-power light microscope. Subsequently, the other part of the soil was processed to isolate total DNA, from which the 18S rRNA gene was sequenced for the identification of nematode taxa. The extracted DNA was also used for randomly amplified polymorphic DNA (RAPD) fingerprinting analysis to determine banding patterns that could classify different bog habitats based on PPN random primers. Compared to that in the climax habitats (HBH, HBL, DBH, DBL, FP, R-FP), PPN prevalence was recorded as being higher in grasslands (WP, RG-I, IFPG-RW and IFPG-NRW) and scrub/woodland ecosystems (C-RW, C-NRW, W-RW, W-NRW). The results indicate that nematode populations are different across the various bog habitats. Emerging and current quarantine PPN belonging to the families Pratylenchidae, Meloidogynidae, Anguinidae and Heteroderidae were noted to be above the threshold limits mentioned under EPPO guidelines, in grassland and wooded peatland habitats. Future actions for PPN management may need to be considered, along with the likelihood that these PPN might impact future paludiculture and other crops and trees growing in nearby agricultural lands.

Title: A simple and efficient protocol for extraction of nematodes from the Irish peat soilsAnusha Pulavarty1, Tilman Klappauf1, Ankit Singh1, Douglas McMillan2,Thomais Kakouli-Duarte1 Molecular Ecology and Nematode Research Group, enviroCORE,Department of Applied Science, South East Technological University, Kilkenny Road Campus, Carlow1Green Restoration Ireland Cooperative Society Ltd (GRI), Ireland2Corresponding author: Anusha.pulavarty@setu.ie Abstract Natural bogs and peatlands play a major role in tackling the climate crisis, as they are natural carbon sinks when intact, but are significant carbon sources when drained. Over 80% or 1.2 million hectares of the 21% of Ireland's land area, which is peatland, is in a degraded condition and so is emitting millions of tonnes of CO2 annually. This makes it a significant, if not the biggest, single source of carbon pollution in the country. This degradation also causes water pollution and disturbs the flora and fauna of these endangered ecosystems. Numerous rewetting and restoration programs have been implemented throughout Ireland and Europe with the goal of rehabilitating these valuable habitats. The enterprise partner in this project, Green Restoration Ireland (GRI) cooperative, is working directly to re-establish this key component of Ireland's natural heritage, with a particular focus on peat grasslands, in order to help fight climate change and restore peatland biodiversity, while diversifying incomes for landowners. The SETU’s Molecular Ecology and Nematode Research Group are assisting GRI in evaluating their peatland restoration programme by studying the nematode diversity in intact, restored and non-restored peatlands. Overall, 14 different peatland habitats including raised bogs and fens have been sampled to extract nematodes from the peat soils. For nematode identification, both morphological and molecular approaches have been used. For morphological identification of the nematodes, their efficient extraction from the peat soils is crucial. Existing protocols in the literature are time consuming and cumbersome. Therefore, a quick and easy extraction procedure has been developed and standardised to extract nematodes from different peatland habitats. The various peat habitats in this study have been classified as: healthy bog hummock (HBH), healthy bog lawn (HBL), degraded bog hummock (DBH), degraded bog lawn (DBL), wasted peat (WP), rough grazing (RG-I), cutover scrub- rewetted (C-RW), cutover scrub non-rewetted (C-NRW), woodlands rewetted (W-RW), woodlands non-rewetted (W-NRW), fen peat (FP), improved fen peat grasslands rewetted (IFPG-RW), improved fen peat grassland non-rewetted (IFPG-NRW) and rich fen peat (R-FP). This protocol could provide time efficiency by significantly reducing the effort of nematologists and soil ecologists working on morphological identification of nematodes from peatlands. Keywords: nematodes, peatlands, bogs, extraction, carbon sinks Graphical overview Background:According to the Irish Peatland Conservation Council (IPPC 2012), boglands in Ireland are special habitats occupying up to 21% of the Irish landscape (O’Connell, 2012) and support many rare plants and animals of national and international importance (Kearns, 2018). Raised bogs started forming 10,000 years ago when much of central Ireland was covered by shallow lakes left behind by the melting ice. The peat in bogs was formed by the accumulation of layer upon layer of partly decomposed dead plants over millenia in these waterlogged regions (Blackith and Speight, 1974). Peatlands play a significant role as global atmospheric carbon storage areas as they are natural carbon sinks but when drained are significant carbon sources. The global carbon cycle, and ultimately climate change, are therefore greatly influenced by the uptake of carbon dioxide (CO2) by these lands (Watson and O'Hare, 1973). Peatlands have been exploited for peat extraction and agriculture for centuries, therefore, over 80% of Irish peatlands are in a degraded state. Approximately 1.2 million of Ireland’s 1.5 million hectares of peatlands are damaged to different degrees. This disturbs the natural flora and fauna of these ecosystems and releases sequestered CO2 back into the atmosphere contributing to climate change. Numerous rewetting and restoration programs have been implemented throughout Ireland and Europe with the goal of rehabilitating degraded bogs.The enterprise partner and co-funder of the project Green Restoration Ireland Coop (GRI) is actively working towards restoration of the degraded bogs. GRI are developing hands-on technology in the form of an app, to be provided to farmers with agricultural peatlands as a tool to enable better management that will reduce environmental impacts and greenhouse gas emissions, in particular. The European Innovation Partnerships (EIP) scheme supports projects that allow farmers, scientists and other experts to collaborate for the development of new practices that are environmentally friendly and economically sustainable. The ultimate aim of EIPs is to road-test new ideas and practices which can then be used more widely by farmers and others to improve productivity and enhance resource efficiency. The enterprise partner is working to optimise this scheme in Ireland, in relation to peat grasslands. The Molecular Ecology and Nematode Research Group (MENRG) in enviroCORE, SETU, has been invited to collaborate with GRI on this, bringing and applying nematode biomonitoring expertise to confirm that GRI’s approaches are successful in restoring these peatlands ecosystems. Nematodes are excellent environmental bioindicators because they are (a) highly abundant and diverse, (b) easily sampled and sorted, (c) representative of their habitat, be it terrestrial or aquatic and (d) known to exhibit well-defined responses to environmental challenges (Wilson and Kakouli-Duarte, 2009). Therefore, this research...

Title: A simple and efficient protocol for extraction of nematodes from the Irish peat soilsAnusha Pulavarty1, Tilman Klappauf1, Ankit Singh1, Douglas McMillan2, Thomais Kakouli-Duarte1 Molecular Ecology and Nematode Research Group, enviroCORE,Department of Applied Science, South East Technological University, Kilkenny Road, Carlow1Green Restoration Ireland Cooperative Society Ltd (GRI), Ireland2Correspondence author: Anusha.pulavarty@setu.ie Abstract Natural bogs and peatlands play a major role in tackling the climate crisis, as they are natural carbon sinks when intact, but are significant carbon sources when drained. Over 80% or 1.2 million hectares of the 21% of Ireland's land area, which is peatland, is in a degraded condition and so is emitting millions of tonnes of CO2 annually. This makes it a, significant, if not the biggest, single source of carbon pollution in the country. This degradation also causes water pollution and disturbs the flora and fauna of these endangered ecosystems. Numerous rewetting and restoration programs have been implemented throughout Ireland and Europe with the goal of rehabilitating these valuable habitats. The enterprise partner in this project, Green Restoration Ireland (GRI) cooperative, is working directly to re-establish this key component of Ireland's natural heritage to help fight climate change and restore peatland biodiversity, while diversifying incomes for landowners. The SETU’s Molecular Ecology and Nematode Research Group are assisting GRI in evaluating their peatland restoration programme by studying the nematode diversity in intact, restored and non-restored peatlands. Overall, fourteen different peatland habitats including raised bogs and fens have been sampled to extract nematodes from the peat soils. For this purpose, both morphological and molecular approaches have been used. For morphological identification of the nematodes, their efficient extraction from the peat soils is very crucial. Existing protocols in the literature are very time consuming and cumbersome. Therefore, a quick, easy and a specific extraction procedure has been developed and standardised to extract nematodes from different peatland habitats. The various peat habitats in this study have been classified as: healthy bog hummock (HBH), healthy bog lawn (HBL), degraded bog hummock (DBH), degraded bog lawn (DBL), wasted peat (WP), rough grazing (RG-I), cutover scrub- rewetted (C-RW), cutover scrub non-rewetted (C-NRW), woodlands rewetted (W-RW), woodlands non-rewetted (W-NRW), fen peat (FP), improved grasslands fen rewetted (IGF-RW), improved grassland fen non-rewetted (IGF-NRW) and rich fen peat (R-FP). This protocol could provide time efficiency by significantly reducing the effort of nematologists working on morphological identification. Keywords: nematodes, peatlands, bogs, extraction, carbon sinks Graphical overview Background:According to the Irish Peatland Conservation Council (IPPC 2012), boglands in Ireland are special habitats occupying up to 21% of the Irish landscape (O’Connell, 2012). These regions accommodate many rare plants and animals of national importance (Kearns, 2018). Raised bogs were formed 10,000 years ago when much of central Ireland was covered by shallow lakes left behind by the melting ice. Bogs were formed by dead plants, which got accumulated layer upon layer over centuries in waterlogged regions (Blackith and Speight, 1974). The CO2 levels in atmosphere are greatly affected by peat accumulation and decay. The peatlands are natural carbon sinks; therefore, climate change and the global carbon cycle are greatly influenced by the uptake of CO2 by these lands (Watson and O'Hare, 1973). Over the past few decades, the peatlands have been exploited for peat extraction. This led to the drastic degradation of 80% of Irish peatlands. Approximately 1.2 million of Ireland’s 1.5 million hectares of peatlands are damaged to different degrees, disturbing the natural flora and fauna of these ecosystems and releasing sequestered carbon dioxide back into the atmosphere contributing to climate change. Numerous rewetting and restoration programs have been implemented throughout Ireland and Europe with the goal of rehabilitating degraded bogs.The enterprise partner and co-funder of the project Green Restoration Ireland Coop (GRI) is actively working towards restoration of the degraded bogs. Green Restoration Ireland Coop are developing hands-on technology in the form of an app, to be provided to farmers with agricultural peatlands as a tool to enable better management that will reduce environmental impacts and greenhouse gas emissions, in particular. The European Innovation Partnerships (EIP) scheme supports projects that allow farmers, scientists and other experts to collaborate for the development of new practices that are environmentally friendly and economically sustainable. The ultimate aim of EIPs is to road-test new ideas and practices which can then be used more widely by farmers and others to improve productivity and enhance resource efficiency. The enterprise partner is working to optimise this scheme in Ireland, in relation to peat grasslands. EnviroCORE’s Molecular Ecology and Nematode Research Group (MENRG) has been invited to collaborate with them on this, bringing nematode biomonitoring expertise to confirm that their approaches are successful in restoring these peatlands ecosystems. Nematodes are excellent environmental bioindicators because they are (a) highly abundant and diverse, (b) easily sampled and sorted, (c) representative of their habitat, be it terrestrial or aquatic and (d) known to exhibit well-defined responses to environmental challenges (Wilson and Kakouli-Duarte, 2009). Therefore, this research will analyse nematode biodiversity as a means of assessing the ecological status of the restored sites. Similar studies have previously been performed, where nematode communities have been used as...