Environmental Resource Management · Deoxynivalenol (DON) is a toxic compound produced by cereal-pathogenic Fusarium fungi. DON is harmful to both animal and plant tissues. Evidence

FACULTY OF AGRI-FOOD AND THE ENVIRONMENT RESEARCH REPORT 2002-2003 87

The Department of Environmental Resource Managementoffers a range of possibilities for postgraduate studentswishing to work in environmental science. Our postgrad-uate research students come from a wide range of primarydegree backgrounds. They work closely with our postdoc-toral researchers and with them, make a major contribu-tion to our research effort. Our current research interestsinclude forest ecosystem studies, land use and landscapeplanning; environmental resources and rural tourism,ecology and rehabilitation of peatlands; nutrient utilisationand losses from agricultural land; trace gas emissions frompeatlands and mineral soils; wildlife ecology andmanagement; role of fauna in agroecosystems; biology,biodiversity of managed ecosystems, management andcontrol of a range of crop pests and pathogens and animalparasites; genetic diversity of plant and microbialpopulations, programmed cell death in plants, molecularplant pathology, epidemiology of diseases in crops andlivestock; environmental effects of pesticides.

Our research is supported by many fundingagencies, including the EU Fifth Framework Project(FP5), the Wellcome Trust Research Fund, ScienceFoundation Ireland, Research Stimulus Fund,Environmental Protection Agency, COFORD, Teagasc,the Forest Service and the Higher Education Authority.

We have strong research programmes on conserva-tion and biodiversity, environmental biotechnology,forest ecosystems, nutrient management, soil ecology,pest and disease management, parasitic and zoonoticinfections and wildlife management. We are continuallybuilding on these and other research programmes,increasing our understanding of ecosystems, contribut-ing to the resolution of regional and national problems,providing opportunities for research students andenhancing our own performance as educators.

The Department offers opportunities for postgradu-ate study in a range of disciplines related to agricultureand other forms of land use and the naturalenvironment. Interdisciplinary MSc(Agr) programmesin Environmental Resource Management and PlantProtection are available as well as single-disciplineMAgrSc, MSc(Agr) and PhD options in AgriculturalBotany, Agricultural Zoology, Plant Pathology, ForestEcology, Plant and Microbial Molecular Biology and SoilScience. A wide range of equipment and facilities areavailable for plant, animal and soil research. Theseinclude facilities for field work at Lyons Estate,laboratory, controlled-environment and glasshousefacilities for plant propagation and for the isolation andmaintenance of plant and animal pathogens andparasites; serological, PCR and other diagnostictechniques; ELISA plate reader; laminar flow cabinets;molecular biology and cell culture equipment; light,interference phase and fluorescence microscopy;radiotelemetry for wildlife tracking; ICP; UV andatomic absorption spectrophotometry; automated col-orometric analysis; infrared gas analysis; gas chro-matography; X-ray fluorescence; density gradient frac-tionation; electrophoresis, ultra/refrigerated centrifuge,image-analysis equipment and a wide range ofcomputing equipment.

The Department has a good record in publishing theresults of our research in peer-reviewed journals ofinternational significance. This provides a valuableopportunity for postgraduate students and post-doctoral researchers alike to build up strong researchportfolio. The following pages give a flavour of theresearch we have been doing. Our aim is to provide ascientific base for the sustainable management of therural environment.

Environmental ResourceManagement

StaffAcademic Staff: Professor Edward P. Farrell (Head), Dr Kevin J. Clancy, Professor Bryan M. Cooke,Professor James P. Curry, Dr Fiona Doohan, Dr John Feehan, Dr David J. Fry, Professor Jeremy S. Gray, Dr Tamara Hochstrasser, Dr Grace O’Donovan, Dr Patrick O’Toole, Dr Gordon Purvis, Dr Michael L. Reilly, Dr Olaf Schmidt, Professor John Whelan

Post-doctoral Researchers: Dr Annette Anderson, Dr Josephine Brennan, Dr Roy Browne, Dr AlvinHelden, Dr Suzanne Monaghan, Dr Sharon Parr, Dr Jane Tuohy

Forest Ecosystem Research Group Staff: Mr Norman Butler, Ms Carly Green, Ms Minna Pollanen, MsFlorence Renou

Technicians: Mr David Byrne, Mr Damian Egan, Mr Brian Fagan, Ms Erica Guerts, Mr Bernard Kaye, Mr Gerald Leonard, Ms Gillian McGrath, Mr Rae Sullivan

Executive Assistants: Ms Helen McCarthy, Ms Jeanette Purvis

Telephone: 716 7737 Fax: 716 1102 E-mail: [email protected] staff e-mails at: http://www.ucd.ie/e-mail/a.html

FACULTY OF AGRI-FOOD AND THE ENVIRONMENT RESEARCH REPORT 2002-200388

ENVIRONMENTAL RESOURCE MANAGEMENT

Trichothecenes are chemical compounds calledsesquiterpene epoxides and are produced byseveral phytopathogenic Fusarium fungal species.They inhibit protein synthesis in various higherorganisms and have been linked to both animaland human poisoning (mycotoxicosis) [1]. Of thenumerous trichothecenes, deoxynivalenol (DON)is commonly produced by several plant-pathogenic Fusarium species causing Fusariumhead blight (FHB) disease of cereals [2]. DON istoxic to plants (phytotoxic) and its productionincreases the virulence of Fusarium graminearumtowards wheat [3]. This study investigated the roleof DON on gene expression in roots of differentwheat cultivars.

Materials and methodsGerminating seedlings (20°C, 48 h old) of wheatcvs. Frontana, CM 82036, Remus and Riband werevacuum infiltrated (20 min) with, and furthergerminated (10°C) on, either 20 ppm DON orwater and harvested 4, 24 or 48 h post-treatment.Gene expression in root samples harvested after 24h was assessed by a molecular technique calleddifferential display reverse transcriptasepolymerase chain reaction (DDRT-PCR) analysisof RNA extracts [4,5,6]. Differentially expressedbands were re-amplified as above, cloned (pGEM-T® cloning kit, Promega, UK), sequenced (MWGBiotech, Germany), and their homology wasdetermined using the NCBI blast search engine[7]. DDRT-PCR results were confirmed, and thetemporal expression of the transcripts (4, 24 and 48h post treatment) analysed, by a technique calledRT-PCR [5] of RNA extracts (1µg per reaction)

using gene-specific and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)-specificprimers (as a control, constitutively expressedgene). Statistical analysis of RT-PCR data wasconducted using the Mann-Whitney Wilcoxon runsum test (SPSS v8.02 software).

Results and discussionDDRT-PCR identified seventy bands over-expressed in DON compared to water-treatedwheat roots of all four cultivars tested harvested24 h post-treatment. Eleven bands were re-amplified, cloned and sequenced (Table 1).

Gene-specific RT-PCR analysis showed thatthe translation elongation factor 1α (EF-1α)homolog (C20B1) was significantly overex-pressed due to DON treatment in roots ofcultivars Frontana, Remus and Riband at alltime-points and in CM 82036 at 24 and 48 h post-treatment (P<0.05) (Fig. 1). In contrast, theadenosine kinase (ADK) (C20B5) expression wassignificantly suppressed in the roots of Frontanaat 24 and 48 h post-treatment and in the otherthree cultivars at all time points (P<0.05). Boththese two house-keeping genes participate indiverse biochemical processes from cell wallformation to programmed cell death [8,9,10]. Up-regulation of EF-1a under stress condition wasreported earlier [9], while down regulation ofADK expression in ADK-deficient mutants ofArabidopsis thaliana plants showed numerousmorphological changes due to inhibition of S-adenosyl-L-Met (SAM)-dependent transmethy-lation reactions needed for proper development[10].

Genotype-dependent accumulation of wheat tran-scripts in response to a fungal mycotoxinK. I. Ansari, D. Egan and F. M. Doohan

Deoxynivalenol (DON) is a toxic compound produced by cereal-pathogenicFusarium fungi. DON is harmful to both animal and plant tissues. Evidence suggestthat DON acts as a virulence factor and helps Fusarium during infection of host tis-sue. We used molecular techniques to determine which plant (wheat) genes respondto DON by analysing the accumulation of RNA transcripts of genes. Results showedthat at least 70 RNA transcripts were over-produced in response to this toxin. Furtheranalysis showed that eleven transcripts responded to toxin treatment in a time- andwheat cultivar-dependant manner.



The Romani retrotransposon homolog (C20B10)was significantly up-regulated in cultivars Frontanaand CM 82036, but not in Remus and Riband, inresponse to DON (P<0.05). The Erika long terminalrepeat (LTR) homolog (C20B6) was significantly up-regulated by DON in roots of cultivars Frontana,CM 82036 and Remus either at 4, 24 and/or 48 hpost-treatment, but not in cv. Riband (P<0.05).Expression of retrotransposons due to DONtreatment supports the previous finding that retro-transposons may play a positive role by acting as aDNA repair mechanism and increasing geneticvariation under stress conditions [11].

DON up-regulation of several genes of unknownfunction (C4B, C4C and C4D) was significantlyhigher in cultivars Frontana and CM 82036 (234 to550%) than in Remus and Riband (126 to 225%)(Figure 2). DON treatment resulted in up-regulationof other genes of unknown function, includingC20B14 in all cultivars and delayed up-regulation (48h post-treatment) of C4C4 in cultivars Frontana, CM82036 and Remus (232 to 329%), but not in Riband(123%). Only a subtle up-regulation of C4A (127%)was observed due to DON treatment (results not

Transcript code Genotype-specific overexpression (> 130 %) asdetermined by RT-PCR or northern blot analysis

Nucleic acid homologs: total transcript sequence lengthand percentage homology in a given overlap (nt)

C20B1 All Translation elongation factor : 658 nt, 93% homologyin 532 nt overlap

C20B5 None (underexpressed in all by 25 to 35 %) Rice adenosine kinase: 483 nt, 90% homology in a 368nt overlap

C20B6 Frontana Long terminal repeat (LTR) of wild wheat (Triticummonococcum) Erika retrotransposon: 353 nt, 79%homology in a 316 nt overlap

C20B10 Frontana and CM 82036 Polyprotein of wild wheat Romani retrotransposon(Gypsy family): 275 nt, 86% homology in a 269 nt overlap

C20B14 All cDNA from Erysiphe graminis-inoculated wheat: 174nt, 100 % homology in 174 nt overlap

C4A None (102 – 115 % only) No significant homology: 436 nt

C4B Frontana and CM 82036 Barley cDNA: 385 nt, 84 % homology in a 271 nt overlap

C4C Frontana, CM 82036 and Remus No significant homology : 333 nt

C4D All cDNA of wheat: 283 nt, 89% homology in a 244 ntoverlap

C4C4 None (Riband = 110 %) cDNA of wheat: 399nt, 94% in a 206 nt overlap

B6B Frontana LTR of T. monococcum Sukkula retrotransposon(unknown family): 294 nt, 90% homology in a 55 and184 nt overlap

Table 1. Transcripts differentially expressed in wheat cultivars Frontana, CM 82036, Remus and/orRiband roots due to DON treatment.

Fig. 1. Effect of deoxynivalenol (DON) on the expression ofthe putative gene C20B1 (EF-1α) over time (4, 24 and 48 hpost-treatment), relative to the expression of GAPDH, inwheat cultivars Remus, Frontana CM 82036 and Riband. (A)Visualisation of C20B1 and GAPDH RT-PCR products.Lanes represent RNA extracts from samples harvested 24 hpost-treatment and treated with: 1–4, water; 5–8 DON.Arrows indicate GAPDH (383bp) and C20B1 (310bp)products. (B) DON relative to water-treated samples. Barsindicate standard error.



shown). Although the number of wheat cultivarsused in this study was limited, these transcripts ofunknown function will be further investigated todetermine if they play any role in a genotype-specificDON response or in host disease resistance.

AcknowledgementsThe FUCOMYR project is funded under the EU 5th

Framework Programme.

References1. Desjardins AE, Hohn TM and McCormick SP, 1993.

Trichothecene biosynthesis in Fusarium species:chemistry, genetics and significance. MicrobiologicalReviews 57, 595–604.

2. Bottalico A and Perrone G, 2002. Toxigenic Fusarium species andmycotoxins associated with head blight in small-grain cerealsin Europe. European Journal of Plant Pathology 108, 611–624.

3. Bai GH, Plattner R., Desjardins A and Kolb F, 2001.Resistance to Fusarium head blight and Deoxynivalenolaccumulation in wheat. Plant Breeding 120, 1–6.

4. Chang S, Puryer J and Cairney J, 1993. A simple and efficientmethod for isolating RNA from pine trees. Plant MolecularBiology Reporter 11, 113–116.

5. Liang P and Pardee AB, 1992. Differential display ofeukaryotic messenger RNA by means of polymerase chainreaction. Science 257, 967–971.

6. Doyle K, 1996. DNA sequencing. In: The sources for discovery,protocol and application guide, K. Doyle (ed.), Promegacorporation, Madison, USA, pp. 147–162.

7. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z,Miller W and Lipman DJ, 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database searchprograms. Nucleic Acids Research 25, 3389–3402.

8. Riis B, Rattan IS, Clark BFC and Merrick WC, 1990.Eukaryotic protein elongation factors. Trends in BiochemicalScience 15, 420–424.

9. Duttaroy A, Bourbeau D, Wang XL and Wang E, 1998.Apoptosis rate can be accelerated or decelerated by over-expression or reduction of the level of elongation factor-1alpha. Experimental Cell Research 238,168–176.

10. Moffatt BA, Stevens YY, Allen MS, Snider JD, Pereira LA,Todorova MI, Summers PS, Weretilnyk EA, McCaffreyLM and Wagner C, 2002. Adenosine kinase deficiency isassociated with developmental abnormalities and reducedtransmethylation. Plant Physiology 128, 812–821.

11. Wessler SR, Bureau TE and White SE, 1995. LTR-retro-transposons and MITEs: important players in theevolution of plant genomes. Current Opinion in Genetics &Development 5, 814–821.

Fig. 2. Effect of deoxynivalenol (DON) on the expression of thetranscript C4B (Barley cDNA) over time (4, 24 and 48 h post-treatment), relative to the expression of GAPDH, in wheatcultivars Remus, Frontana CM 82036 and Riband. (A)Visualisation of C4B and GAPDH RT-PCR products. Lanesrepresent RNA extracts from samples harvested 24 h post-treatment and treated with: 1–4, water; 5–8 DON. Arrowsindicate GAPDH (383bp) and C4B (210 bp) products. (B) DONrelative to water-treated samples. Bars indicate standard error.

Analysis of intra- and inter-species genetic diversityof European wheat-pathogenic Fusarium fungiJ. M. Brennan, D. Egan and F. M. Doohan

Using a molecular technique called AFLP analyses, we examined the intra- and inter-species genetic diversity of eighty wheat-pathogenic Fusarium fungal isolates represent-ing five species (F. avenaceum, F. culmorum, F. graminearum, F. poae and M. nivale) andoriginating from different European climatic regions including the maritime northwest(the UK and Ireland), the Mediterranean Italian and the continental Hungarian climates.Cluster analysis of AFLP data revealed that F. graminearum, F. culmorum and F. poaeshowed greater intra-species genetic similarity (s ≥ 0.63) than F. avenaceum and M. nivale(s ≥ 0.10 and 0.37, respectively). Fusarium avenaceum was the most genetically distinctspecies (s = 0.10 to 0.95), and showed low similarity to the other species (s = 0.10 to 0.44).Principal coordinate analysis confirmed the same general clustering profiles as did thedendrogram. The present work also indicated that, within some species, there may be arelationship between genetic diversity and country of origin of the isolates.



Fusarium head blight (FHB) is an importantfungal disease of wheat, barley and maizeworld-wide [1]. FHB has received significantattention because of its impact on yield and grainquality and, more importantly, because of theability of some Fusarium species to contaminategrain with toxic compounds called mycotoxins[2]. Fusarium fungi exhibit an extraordinarydegree of diversity with respect to morphologi-cal, physiological and ecological characteristics.This study used AFLP analysis to determine theinter- and intra-species genetic variation amongeight wheat-pathogenic Fusarium isolates fromdifferent European climates including themaritime northwest (UK and Ireland), theMediterranean Italian and the continentalHungarian climates.

Materials and methodsThe eighty Fusarium isolates used included F.avenaceum, F. culmorum, F. graminearum, F. poae andMicrodochium nivale (formerly known as Fusariumnivale) from Ireland, Italy, Hungary and the UKand nine other F. graminearum isolates represent-ing different geographic lineages (USA, Brazil,New Zealand, China, South Africa, Honduras,Nepal) [3] and isolates of seven other Fusariumspecies. Isolates were identified morphologicallyand by species-specific PCR analysis [4,5,6,7].DNA was extracted from mycelium [8] and AFLPanalysis was conducted using a procedure similarto that described by Vos et al. [9]. All five selectiveAFLP analyses were conducted twice. AFLPprofiles were analysed using Image Master 1DElite v.3.01 software (Amersham PharmaciaBiotech AB, Sweden). Resulting binary data wereused to calculate the Jaccard similarity coefficients.The PAUP 4.0b10 software package was used tocluster the isolates by the unweighted pairedgroup method with arithmetic averages (UPGMA)with 1000 bootstraps. PCA analysis wasperformed using NTSYS sp 2.0 (Exeter software,New York, USA).

Results and discussionBoth UPGMA cluster analysis of the Fusariumisolates (Figure 1) and principal coordinateanalysis (results not shown) confirmed the samegeneral clustering profiles of the Fusarium isolates.Fusarium graminearum, F. culmorum and F. poaeisolates showed the highest level of genetic

similarity (s ≥ 0.69, 0.63 and 0.63, respectively)compared to F. avenaceum and M. nivale (Figure 1),but, even within these species, there were distinctgroups, agreeing with the work of previousresearchers [3,10,11]. The variation within the F.graminearum, F. avenaceum and M. nivale isolates inthe current study (Figure 1) may be, in part, due tothe fact that these species have a teleomorphicstage, i.e. they can reproduce sexually as well asasexually. As F. culmorum and F. poae have noknown teleomorph, variation might have arisenthrough point mutations, gene flow or recombina-tion [11]. The most genetically diverse specieswere F. avenaceum and M. nivale (s ≥ 0.1 and 0.37,respectively). However, the techniques used toclassify the isolates in this study (i.e. morphologyand species-specific PCR) may not differentiate F.avenaceum from the closely related F. tricinctumspecies [7].

Surprisingly, the fourteen European F.graminearum isolates did not cluster with the F.graminearum geographic lineages (s = 0.42) (Figure1). This differentiation may be due to the non-European geographic origin of the lineages. Afterinvestigations on anamorphic and teleomorphiccharacteristics of F. graminearum isolates, Aoki andO’ Donnell [12] suggested that F. graminearumGroup 1 is a separate species from Group 2 andshould be renamed F. pseudograminearum. Such adistinction could account for some of the variationobserved in the present work.

The present work also indicated that, withinsome species, there might be a relationshipbetween genetic diversity and country of originof the isolates. This was particularly evident forthe fifteen M. nivale isolates that originated fromIreland, the UK and Italy. Those from the UK andIreland clustered together in the AFLP UPGMAdendrogram (s ≥ 0.36); those from Italy formed adistinct group (Figure 1). While the Hungarianisolates were generally the most homogeneous,surprisingly the Irish isolates were the most het-erogeneous. The similarity amongst theHungarian isolates could be due to year ofisolation, similar management practices,sampling location or possibly the uniformity ofthe climate within the country. As the hetero-geneity of the Irish isolates is not due todifferences in year of isolation (2000), it may beassociated with the intercontinental movement ofinfected seed.



Fig. 1. Dendrogram of the Fusarium isolates based on UPGMA cluster analysis using the Jaccard similarity coefficients generatedfrom five selective AFLP profiles. a Isolates are colour-coded according to species: F. avenaceum, F. culmorum, F. graminearum,F. poae and M. nivale. b European isolates. c Origin of the isolates is colour selective coded according to country: Ireland, UK, Italyand Hungary. d ND, no data. Bootstrap values greater than 50% are shown.



AcknowledgementsThe work was funded by the EU 5th FrameworkProgramme project RAMFIC (QLRT-1999 31517).The authors thank Paul Nicholson (John InnesCentre, Norwich, UK), Simon Edwards (HarperAdams University College, Newport, UK),Antonio Logrieco and Antonio Moretti (IstitutoTossine é Micotossine de Parassiti Vegetali, Bari,Italy) and Lazlo Hornok (AgriculturalBiotechnology Center, Gödöllö Hungary) forproviding isolates.

References1. Parry DW, Jenkinson P and McLeod L, 1995. Fusarium ear

blight (scab) in small grained cereals – a review. PlantPathology 44, 207–238.

2. Logrieco A, Mule G, Moretti A and Bottalico A, 2003.Toxigenic Fusarium species and mycotoxins associatedwith Head Blight in small-grain cereals in Europe.European Journal of Plant Pathology 108, 611–624.

3. O’Donnell K, Kistler HC, Tacke BK and Casper HH, 2000.Gene genealogies reveal global phylogeographic structureand reproductive isolation among lineages of Fusariumgraminearum, the fungus causing wheat scab. Proceedings ofthe National Academy of Sciences of the USA 97, 7905–7910.

4. Nicholson P, Lees AK, Maurin N, Parry DW and RezanoorHN, 1996. Development of a PCR assay to identify andquantify Microdochium nivale var. nivale and Microdochium

nivale var. majus in wheat. Physiological and Molecular PlantPathology 48, 257–271.

5. Parry DW and Nicholson P, 1996. Development of a PCRassay to detect Fusarium poae in wheat. Plant Pathology 45,383–391.

6. Nicholson P, Simpson DR, Weston G, Rezanoor HN, LeesAK, Parry DW and Joyce D, 1998. Detection and quantifi-cation of Fusarium culmorum and Fusarium graminearum incereals using PCR assays. Physiological and Molecular PlantPathology 53, 17–37.

7. Turner AS, Lees, AK, Rezanoor HN and Nicholson P, 1998.Refinement of PCR-detection of Fusarium avenaceum andevidence from DNA marker for phenetic relatedness toFusarium tricinctum. Plant Pathology 47, 278–288.

8. Chen D and Ronald PC, 1999. A rapid DNA minipreparationmethod suitable for AFLP and other PCR applications.Plant Molecular Biology Reporter 17, 53–57.

9. Vos P, Hogers R, Bleeher M, Reijans M., Van de Lee T,Peleman, J, Kulper, M and Zabeau M, 1995. AFLP: A newtechnique for DNA fingerprinting. Nucleic Acid Research23, 4407–4414.

10. Carter JP, Rezanoor HN, Desjardins AE and Nicholson P,2000. Variation in Fusarium graminearum isolates fromNepal associated with their host of origin. Plant Pathology49, 452–460.

11. Mishra PK, Fox RTV and Culham A, 2002. Restrictionanalysis of PCR amplified nrDNA regions revealedintraspecific variation within populations of Fusariumculmorum. FEMS Microbiology Letters 215, 291–296.

12. Aoki T and O’Donnell K, 1999. Morphological andmolecular characterisation of Fusarium pseudograminearumsp. nov., formally recognised as group 1 population of F.graminearum. Mycologia 91, 597–609.

Development and evaluation of a novel in vitrodetached leaf assay for detecting resistances toFusarium head blight in wheatR. A. Browne and B. M. Cooke

Fusarium head blight (FHB) is the most important disease of wheat worldwide,primarily due to mycotoxins which occur as a result of the fungal infection. Muchresearch and breeding effort is focused on preventing this disease complex and itsassociated problems. However, mechanisms of resistance to the disease are poorlyunderstood. Research is presented on the development and evaluation of an in vitrodetached leaf assay for pre-screening components of partial disease resistances toFHB in wheat. The methods developed are currently being evaluated through col-laborative research in breeding programmes in Europe and the US.

The Fusarium head blight disease complex is oneof the most important diseases of cerealproduction worldwide, causing serious contami-nation of harvested grain with toxic compoundscalled mycotoxins. Losses in the US alone have

been estimated at US$ 3 billion over a six-yearperiod in the 1990s [1]. Although resistance toFHB is partial and is generally accepted to beunder polygenic control, resistant cultivars areseen as the most viable strategy for control of



FHB and associated mycotoxin contamination.However, the components of partial diseaseresistance (PDR) to FHB are poorly understoodand consequently incorporating resistance intoadapted high-yielding cultivars has beenhampered.

A relationship between PDR components in thedetached leaf assay and FHB was first reported [2]across seven commercial cultivars of wheat usingMicrodochium nivale, an important member of theFHB disease complex. The objectives of thepresent research were to develop and evaluate thedetached leaf assay for use in pre-screeningjuvenile plants for FHB resistance.

Materials and methodsTo facilitate the development and evaluation of thedetached leaf assay, a technique to improvesporulation of M. nivale in culture and to producemycelium-free conidial suspensions was developedusing cellophane-covered potato dextrose agar(CPDA). Full details of the methodology used havebeen reported elsewhere [3].

In the detached leaf assay, once a suitableprotocol was developed [4], twenty wheatcultivars with a range of FHB resistance wereselected from the UK recommended list 2002.The wheat genotypes were grown for 14 daysunder controlled environment conditions [4].The mid-section of the first leaf was mounted onwater agar amended by kinetin (used as asenescence retarder), inoculated with a 10µldroplet of a conidial suspension of M. nivale andincubated at 10°C under a diurnal cycle of nearultra-violet and white light. Assessment ofsymptom appearance and sporulation werecarried out daily under a compound microscope(magnification 40X). The three components ofPDR measured here were: incubation period(period from inoculation to first appearance ofsymptoms under macroscopic examination),latent period (period from inoculation tosporulation) and lesion length (measured on day7 as the necrotic area visible by placing the Petridishes over a light box). Each observation wasthe mean of 80 observations (20 Petri dishes Xfour leaf segments) with three replications.Correlations between PDR components and thesemi-quantitative UK 2003 recommended listratings were calculated using Spearman rankcorrelations in StatView.

Results and discussionThe new method using CPDA gave improvedsporulation of M. nivale and provided mycelium-free conidial suspensions suitable for inoculationin the detached leaf assay. The cellophane disccould be removed from the agar surface with thecolony intact (Figure 1).

In the detached leaf assay, greater delays in thefirst appearance of symptoms (incubation period)and sporulation (latent period) on wheat hostswere correlated to greater FHB disease resistance(UK 2003 recommended list ratings; higher ratingsindicate greater FHB resistance) (rs = 0.53, P < 0.05,Figure 2A; rs = 0.70, P < 0.01, Figure 2B). Thesefindings indicate that PDR components detectedin the detached leaf assay share a common geneticbasis with much of the resistances to FHBexpressed in the head of these wheat cultivars andcould therefore be useful for pre-screeningresistance to FHB. The precise role of theindividual PDR components detected in thedetached leaf assay and whole plant FHBresistance is unclear although the strongest rela-tionship was between FHB resistance and latentperiod.

The detached leaf assay is currently beingevaluated in collaborative work with wheatbreeders in Europe and the US. In addition, otherresistances to FHB that are not detected using thedetached leaf assay are being investigated.

Fig. 1. Sporulating colony of Microdochium nivale var. majusisolate Dard1/M on cellophane membrane removed from thePDA surface after 7 days under a diurnal cycle of NUV andwhite light at 20ºC.



AcknowledgementsThis work presented was supported in part by theEU funded FUCOMYR project, contract QLRT-2000-02044, ‘Novel tools for developing Fusarium-resistant and toxin free wheat for Europe’.

References1. Windels CE, 2000. Economic and social impacts of Fusarium

head blight: Changing farms and rural communities inNorthern Great Plains. Phytopathology 90, 17–21.

2. Diamond H and Cooke BM, 1999.Towards the developmentof a novel in vitro strategy for early screening of Fusariumear blight resistance in adult winter wheat plants.European Journal of Plant Pathology 105, 363–372.

3. Browne RA and Cooke BM, 2004a. A new method forproducing mycelium-free conidial suspensions fromcultures of Microdochium nivale. European Journal of PlantPathology 110, 87–90.

4. Browne RA and Cooke BM, 2004b. Development andevaluation of an in vitro detached leaf assay for pre-screening resistance to Fusarium head blight in wheat.European Journal of Plant Pathology 110, 91–102.

Fig. 2. Incubation period (A), latent period (B) in the detached leaf assay plotted against UK 2003 FHB ratings of thecommercial cultivars.

A B

Potential of bacteria to control Fusarium diseases ofcerealsM. R. Khan, S. Fischer, D. Egan and F. M. Doohan

One hundred and fifty bacterial isolates collected from Irish cereal fields were test-ed against cereal-pathogenic Fusarium species. On growing bacteria and thepathogen together on the same plate, ten bacterial isolates were found to be antago-nistic to Fusarium culmorum, F. graminearum and F. poae. Of these ten bacterial iso-lates, Pseudomonas fluorescens strain MKB 90 and strain MKB 156, as well as thirteenother bacteria, which did not inhibit Fusarium growth directly, reduced F. culmoruminfection on germinating wheat (cultivar GK-Othalom) seeds. Seven of these fifteenbacterial isolates inhibited Fusarium seedling blight disease to varying degrees onall wheat cultivars tested (Access, Marshal, Claire, Soisson and Baldus). Seedlingblight tests showed that Pseudomonas sp. strain MKB 158 reduced F. culmorum infec-tion of wheat and barley stem bases by 79% and 82%, respectively, relative toFusarium-infected control seedlings.

IntroductionFusarium fungi, including the species Fusariumculmorum, F. graminearum, F. poae and F. avenaceumcause seedling blight, foot rot and head blightdiseases of cereals, resulting in yield loss [1].Fusarium head blight (FHB) is a major cereal disease

worldwide [2], which can result in fungal toxin(mycotoxin) contamination of grains. Amongseveral mycotoxins produced by Fusarium species,deoxynivalenol (DON) is the most important,causing diseases in animals and human beings.Chemical control of this disease using pesticides is



inconsistent and breeding for resistance is not yetfully exploited [1]. An alternative, environmentallyacceptable means of controlling Fusarium diseasesmay be the use of non-pathogenic microbes, astrategy called ‘biological control.’ In this study, onehundred and fifty bacterial isolates collected fromIrish cereal fields were evaluated for their ability toinhibit in vitro growth and seedling blight diseasecaused by Fusarium species.

MethodologyBacteria used in this study originated from leavesand roots of field-grown cereals and are held in theUCD Molecular Plant Microbe Interactions Groupmicrobial collection. Some were identified using amolecular technique based on partial 16S rDNAsequences [3]. The Fusarium isolates used includedF. culmorum strain FCF 200, F. graminearum strainHUGR9 and F. poae strain HUPO3 from the UCDFusarium collection. Dual culture plate tests wereperformed using the method described byPedersen et al. [4].

For in vitro laboratory seedling blight tests, air-dried, surface-sterilised wheat seeds (1% v v-1

sodium hypochloride for 10 min) were immersed in48 h old Luria broth bacterial cultures (OD3.0), air-dried, and the ability of F. culmorum to retard coleop-tile (shoot) growth of germinating seeds was deter-mined using an in vitro seedling blight test [5]. For invivo growth chamber seedling blight tests, the stembases of 2-week-old seedlings of wheat (cv. GK-Othalom) and barley (cv. Lux) were inoculated with400µl of a F. culmorum conidia suspension (1x106

spores ml-1 0.2% Tween 20) and 400µl of 48 h-oldbacterial cultures (OD3.0) as described by Doohan etal. [6]. Plants were grown in a climate controlledgrowth room at day/night temperatures of 20/12°Cwith a 12h light period (700µmol m-2 s-1) and

constant humidity of 95%. Visible stem base diseasesymptoms were scored 21 days after inoculationusing the system described by Goulds and Polley [7].

Results and discussionOf the one hundred and fifty bacterial isolatestested, ten isolates suppressed the growth of F.culmorum, F. graminearum and F. poae in dual culturesplate tests (Figure 1 and Table 1). Two of these ten(i.e. Pseudomonas fluorescens strains MKB 90 andMKB 156) also suppressed shoot growth retardationcaused by F. culmorum in in vitro seedling blight tests(Fig. 2 and Table 1). However, the ability of thirteenbacterial isolates to inhibit Fusarium seedling blightin vitro, although they showed no activity againstFusarium in the dual culture tests (Table 1) indicatesthat such bacteria control the disease by means otherthan direct inhibition [8].

Bacterial suppression of Fusarium seedlingblight in the in vitro test was cultivar-specific(Table 1); this may be due to their differentialability to colonise different cultivars [9]. Sevenbacterial isolates were found to inhibit Fusariumseedling blight in all the wheat cultivars tested(Table 1). In vivo seedling blight tests revealed thatPseudomonas sp. strain MKB 158 suppressed F.culmorum seedling blight on wheat and barleystems by 79% and 82%, respectively (Figure 3).This bacterium did not directly inhibit the growthof F. culmorum (Table 1) indicating that it suppress-es Fusarium seedling blight on wheat and barleystems by means other than direct inhibition

Future work will determine the ability of theseseven bacteria to control FHB of different wheatand barley cultivars in glasshouse and field trials.Moreover, investigations will be conducted to seeif they can activate host defense mechanismagainst FHB in wheat and barley cultivars.

Fig. 1. Growth inhibition of Fusarium graminearum strainHUGR9 by Pseudomonas flourescens strain MKB156 in dualculture tests. F. graminearum strain HUGR9 grown in theabsence (A) and presence (B) of Pseudomonas flourescensstrain MKB 156.

Fig. 2. Bacterial inhibition of in vitro coleoptile retardation ofgerminating wheat (cv. GK-Othalom) seeds caused by Fusariumculmorum strain FCF 200. Non-treated (A) and Pseudomonas sp.strain MKB158-treated (B) seeds were germinated on F.culmorum strain FCF 200-inoculated potato dextrose agar plates.

A B A B



AcknowledgementsThis work was funded by Irish Department ofAgriculture, Food and Rural DevelopmentResearch Stimulus Fund.

References1. Parry DW, Pettitt TR, Jenkinson P and Lees AK, 1994. The

cereal Fusarium complex. In Ecology of Plant Pathogens, J.P.Blakeman and B. Williamson (eds.). CAB International,Wallingford, pp. 301–320.

2. McMullen M, Jones R and Gallenberg D, 1997. Scab ofwheat and barley: a reemerging disease of devastatingimpact. Plant Diseases 81, 1340–1348.

3. Desjardins E and Beaulieu C, 2003. Identification of bacteriacontaminating pulp and a paper machine in a Canadianpaper mill. Journal of Industrial Microbiology andBiotechnology 30,141–145.

4. Pedersen EA, Reddy MS and Chakravarty P, 1999. Effect ofthree species of bacteria on damping-off, root rot develop-ment, and ectomycorrhizal colonization of Lodgepole pineand White spruce seedlings. European Journal of ForestPathology 29, 123–134.

5. Masterhazy A, 1983. Breeding wheat for resistance toFusarium graminearum and F. culmorum. Zeitschrift fürPflanzenzüchtung 91, 295–311.

6. Doohan FM, Weston G, Rezanoor HN, Parry DWandNicholson P, 1999. Development and use of a reversetranscription-PCR assay to study expression of Tri5 byFusarium species in vitro and in planta. Applied andEnvironmental Microbiology 65, 3850–3854.

7. Goulds A and Polley RW, 1990. Assessment of eyespot andother stem base diseases of winter wheat and winterbarley. Mycological Research 94, 815–822.

8. Pieterse CMJ, Wees SCMV, Hoffland E, Pelt JAV and LoonLCV, 1996. Systemic resistance in Arabidopsis induced bybiocontrol bacteria is independent of salicylic acid accu-mulation and pathogenesis-related gene expression. ThePlant Cell 8, 1225–1237.

9. Germida JJ and Siciliano SD, 2001. Taxonomic diversity ofbacteria associated with the roots of modern, recent andancient wheat cultivars. Biology and Fertility of Soils 33,410–415.

Dual culture tests ( % growth inhibition relative to

control cultures)

In vitro control of Fusarium culmorum strain FCF 200seedling blight of different wheat cultivars

( % coleoptile growth relative to untreated seeds)

IsolateCode

Species* F. culmorum

F. graminearum

F. poae

GKOthalom

Access Marshal Claire Soisson Baldus

MKB 25 ND 0 0 0 47 35 96 30 95 85MKB 41 Exiguobacterium sp. 0 0 0 49 0 0 0 0 0MKB 55 ND 0 0 0 23 0 0 0 0 0MKB 74 Low G+C gram-positive

bacterium D-Su(1)-21B0 0 0 30 0 0 0 0 0

MKB 76 ND 18 12 13 0 0 0 0 0 0MKB 85 Bacillus mycoides 0 0 0 26 0 0 0 0 0MKB 86 ND 44 11 14 0 0 0 0 0 0MKB 90 Pseudomonas fluorescens 33 52 36 58 0 0 0 0 0MKB 99 ND 11 30 13 0 0 0 0 0 0MKB 100 Ps. fluorescens 0 0 0 26 14 49 75 55 30MKB 107 ND 33 29 14 0 0 0 0 0 0MKB 121 ND 0 no 0 56 14 53 60 50 45MKB 142 ND 42 25 10 0 0 0 0 0 0MKB 156 Ps. fluorescens 37 36 42 32 0 0 0 0 0MKB 158 Pseudomonas sp. 0 0 0 42 37 89 55 62 77MKB 159 ND 0 0 0 36 0 0 0 0 0MKB 193 ND 31 31 13 0 0 0 0 0 0MKB 202 Ps. fluorescens 0 0 0 92 25 13 35 25 32MKB 207 ND 30 11 13 0 0 0 0 0 0MKB 242 ND 35 10 11 0 0 0 0 0 0MKB 249 ND 0 0 0 60 29 30 17 25 15MKB 264 ND 0 0 0 31 0 0 0 0 0MKB 277 ND 0 0 0 37 12 40 42 44 15

Table 1. In vitro bacterial inhibition of Fusarium growth and Fusarium seedling blight.

Fig. 3. Bacterial inhibition of Fusarium culmorum strain FCF200 stem base disease of wheat and barley seedlings in vivo.Bacteria: Pseudomonas fluorescens strain MKB156 andPseudomonas sp. strain MKB158.



Tick detachment studies in relation to disease transmissionJ. S. Gray, O. Kahla, M. Kundib and G. Stanekb

aFree University of Berlin, Germany bMedical University of Vienna, Austria

Ticks transmit many diseases to both livestock and humans. Whereas livestock tick-borne diseases can be prevented by the application of chemicals (acaricides) and themanagement of grazings, the best defence against human diseases transmitted bythese parasites is increased awareness and detachment of feeding ticks from theskin, possibly followed by chemotherapy. The management of tick bites, includingmethods of tick detachment and risk assessment of disease transmission, remainscontroversial because of a lack of objective data. The studies described here soughtto relate the risk of transmission of Lyme borreliosis, the most important tick-bornedisease of animal origin (zoonosis) in the temperate northern hemisphere, to themanner and timing of tick detachment.

The tick Ixodes ricinus L. is a common andwidespread arthropod vector of human andanimal diseases in northern Europe (Figure 1).One of the diseases it transmits, Lyme borreliosis(LB), caused by spirochaete bacteria of the Borreliaburgdorferi sensu lato (s.l.) genospecies complex, isthe most prevalent arthropod-borne disease ofhumans in temperate regions of the northernhemisphere, including north America and Asia,where other Ixodes species serve as vectors. Thedifficulties associated with the diagnosis andtreatment of Lyme borreliosis and the highfrequency of tick bites amongst exposedpopulations mean that a risk analysis approach tothe probability of disease transmission andappropriate intervention is necessary. Antibiotictherapy has often been advocated following tickbite, but there are insufficient data on the risk of

infection and subsequent disease in relation tomethods of tick removal and duration ofattachment of the tick to the skin. The presentstudies investigated the influence of differentmethods of tick removal and of duration of tickfeeding on transmission of B. burgdorferi tolaboratory gerbils, and also established the rela-tionship between duration of feeding and thedimensions of ticks detached from laboratoryrabbits, as a means to further assess disease trans-mission risk.

Effect of tick removal method and feedingduration on pathogen transmissionThe debate surrounding the detachment of ticksfrom human skin has mainly concerned theproblems of complete removal of embeddedtick mouthparts, and of avoiding increasedtransmission risk by damaging or irritating thetick. Whereas rapid, aggressive detachment islikely to leave remnants of tick mouthparts inthe bite-lesion, the application of heat orchemicals to encourage the tick to detach, maycause an increase in salivary gland secretionand thus in pathogen transmission. The longerthe tick is attached the more likely it is totransmit pathogens, and American studiessuggest that most transmission occurs approxi-mately 48 hours after the commencement offeeding [1].

Fig. 1. A recently attached female Ixodes ricinus tick in anunusual location (courtesy of Prof. G. Stanek)



These aspects were investigated using gerbils(Meriones unguiculatus) infested with B. burgdorferi-infected nymphal I. ricinus ticks (three per gerbil)confined in 7mm diameter plastic capsules. Theticks were detached using one of three differentmethods:

1) Rapidly by grasping the base of themouthparts as close to the skin as possiblewith a pair of fine forceps and exerting firm,steady traction.

2) Slowly and roughly, involving a 3 min period ofintensive squeezing of the tick with fine forcepsto simulate inefficient and slow removal.

3) After application of nail polish for 1 h to inhibitrespiration by blocking the tick spiracles.

Each group of ticks was further subdividedaccording to duration of feeding and ticks weredetached approximately 17, 29, 47 and 65 h afterattachment. Two weeks after tick detachment eachgerbil was investigated for patent spirochaeteinfection by xenodiagnosis using pathogen-freelaboratory-reared larval I. ricinus. Approximately100 tick larvae were applied with a small paint-brush to each gerbil, the resulting engorged larvaewere collected, allowed to develop to nymphs andthe unfed nymphs were then examined forinfection by indirect immunofluorescent assay.

The results (Table 1) showed that the method oftick removal had no effect on the likelihood oftransmission, indicating that irritation of the tickduring the removal process did not increase therisk of transmission. After 47 h of tick feeding, allgerbils had become infected, but an average of56% of the gerbils had become infected after only17 h of tick feeding, which contrasts strongly withthe American findings [1]. The overall conclusionswere that to minimise the likelihood of infection, aprompt, aggressive approach to tick removal isessential and that the risks of causing problemsthrough irritating ticks or damaging them aresmall or non-existent.

These results informed the design and imple-mentation of a follow-up study that examined therelationship between duration of feeding and thechange in tick dimensions during the feedingprocess. The establishment of such a relationshipcan be used to estimate the length of time that atick has been attached to a patient and thusprovide an indication of infection risk.

Determination of tick feeding durationSince it is anticipated that the data produced bythis study will be used in prospective studies onthe epidemiology of LB in highly endemic regionsof Europe, the nymphal and adult female tickswere collected from areas in Austria (70km northof Vienna) where the incidence of LB approaches150 per 100,000 population.

Two New Zealand White rabbits (Oryctolaguscuniculus) were infested with ticks confined in 5cmdiameter plastic capsules and detached at regularintervals (6, 12, 24, 36, 48, 60 and 72h for nymphs,and 12, 24, 48, 72, and 96h for adult females) usingfine forceps. The degree of engorgement (as ameasure of feeding duration) of detached ticks wasdetermined by measuring the ratio of the width ofthe scutum (the hard sclerotised plate on thedorsum of the tick) to the total length of the body.This ratio is the Scutal Index and was first used forthe main American vector of Lyme borreliosis,Ixodes scapularis [2]. However, in view of therelatively early transmission of B. burgdorferi by theEuropean vector (see above), a secondmeasurement was made to exploit the fact thatbody width dimensions change earlier than bodylength during feeding. To determine body widththe distance between the base (coxa) of the 4th pairof legs was measured. The ratio of thismeasurement to scutal width is the Coxal Index.

Mean hours oftick feeding

Removalmethod

Bb positive ger-bils (6 pergroup) (%)

16.7 Quick 67

Slow 50

Nail-polish 50

28.9 Quick 50

Slow 67

Nail-polish 40

47.0 Quick 100

Slow 100

Nail-polish 100

65.2 Quick 100

Slow 100

Nail-polish 100

70-120 Not removed 100

Table 1. Effect of nymphal tick (Ixodes ricinus)feeding duration and removal method on transmis-sion of Borrelia burgdorferi s.l. (Bb) to gerbilsdetected by xenodiagnosis.



The results showed that the Coxal Index is moresensitive than the Scutal Index for both adult andnymphal I. ricinus over the first 24h of feeding, butafter 48h this advantage had disappeared (Figure2). The relationships between the indices and tickfeeding durations (t) are expressed by thefollowing regression equations:

Scutal Index, nymphslogSI=10.503x10-3 t + 0.576 r2 =0.82

Scutal Index, adults logSI= 6.161x10-3 t + 0.562 r2 =0.76

Coxal Index, nymphslogCI=17.359x10-3 t - 0.751 r2 =0.93

Coxal Index, adultslogCI=15.422x10-3 t - 0.950 r2 =0.94

Feeding duration may best be determined byestimating both the Scutal and Coxal Indices for adetached tick, and applying the appropriateregression equation, depending on whether thetick was in the early or late feeding phase.

Assessment of the relevance of the datapresented here to the transmission of tick-bornepathogens to humans awaits prospective studiesinvolving patients. It is also necessary to obtainmore information on the dynamics of B. burgdorferitransmission by both nymphal and adult ticksover the first 24h of feeding. However, it is evidentthat the use of Coxal and/or Scutal Indices for tickspecimens removed from humans has thepotential to contribute to more accurate riskassessment and prevention of Lyme borreliosis.

References1. Piesman J, Mather TN, Sinsky RJ and Spielman A, 1987.

Duration of tick attachment and Borrelia burgdorferi trans-mission. Journal of Clinical Microbiology 25, 557–558.

2. Falco RC, Fish D and Piesman J, 1996. Duration of tick bitesin a Lyme disease-endemic area. American Journal ofEpidemiology 143, 187–192.Fig. 2. Comparison of Scutal Index (blue) and Coxal Index (red)

as a function of feeding time (four parameter logistic model).

Animal reservoirs of tick-borne zoonoses in EuropeB. Pichon, A. Estrada-Penaa, O. Kahlb, A. Mannellic and J. S. GrayaUniversity of Zaragoza, Spain, bFree University of Berlin, Germany, cUniversity of Turin, Italy

Ticks are among the most important vectors of human and animal disease in the north-ern hemisphere. Identification of reservoir hosts is a prerequisite for effective preven-tion and control of tick-borne zoonoses, but presents many difficulties, such as the needto trap large numbers of wild animals, and to perform laboratory transmission experi-ments. A more efficient approach, developed in this laboratory, consists of the molecu-lar analysis of the blood-meal remnant of ticks infected by the particular pathogen ofinterest. The study presented here describes the use of this approach to investigate thediversity of tick-borne pathogens and their reservoir hosts in different parts of Europe.

Adult Females

Nymphs



The castor bean tick, Ixodes ricinus, is the mostwidespread and common tick in Europe. In addi-tion to being the vector of several animal diseases,it also transmits important human diseases(zoonoses), including Lyme borreliosis (caused byBorrelia burgdorferi s.l.). DNA detection methodswere developed to identify the animal reservoirsof pathogens transmitted by this tick and requireintegration of the identity of zoonotic pathogenswith that of the host origin of blood-meal rem-nants in the same host-seeking ticks. This studydemonstrates the field-use of this approach in dif-ferent European countries and provides newinsights into the ecology of some of the pathogensinvolved.

Materials and methodsUnfed, host-seeking nymphal I. ricinus ticks werecollected from comparable woodlands in Ireland,Germany, Spain and Italy. The ticks were analysedin UCD to detect pathogens and to identify thehosts that the nymphs had fed on as larvae [1].

Briefly, vertebrate DNA in the tick gut was identi-fied by PCR amplification using universal primerstargeting part of the 18S rRNA gene, followed byReverse Line Blot (RLB). This method identifiedhosts at the subgroup level, e.g. ruminants,hares/rabbits, carnivores, rodents, insectivores,game birds, song birds. The pathogens weredetected in the same tick material with a novelmultiplex PCR, and RLB hybridisation was thenperformed to identify them.

ResultsIreland - Ticks were collected from small areas ofdeciduous/coniferous woodland in a parklandsetting on the outskirts of Killarney, Co. Kerry. Atotal of 756 nymphs were analysed for pathogensand 322 for blood meals. The most frequentpathogens detected (18.4%) were members of theBorrelia burgdorferi genospecies complex, the mostcommon of which were B. garinii (43%) and B.valaisiana (29%). B. afzelii and B. burgdorferi s.s. wererelatively uncommon (13 and 11%, respectively).

Ap Babesia spp Ba Bb Bg Bv RFLB Rh No pathogen

A. Ireland

Large mammals 100 0 6.6 0 0 0 0 0 NA

Rodents 0 0 13.3 0 4.2 3.8 0 0 NA

Birds 0 0 20 80 48 68.6 57 0 NA

Unknown 0 100 60 20 45.8 28.6 43 100 NA

B. Germany

Large mammals 100 0 0 0 0 0 NA

Rodents 0 0 16.7 16.7 50 0 33 11.1 NA

Birds 0 0 6.7 16.7 50 50 0 0 NA

Unknown 0 0 76.7 66.7 0 50 66 88.9 NA

C. Spain

Large mammals 0 0 16.7 0 0 0 0 0 36.3

Rodents 0 0 16.7 0 0 0 0 0 18.2

Birds 0 0 50 0 100 100 0 0 45.4

Unknown 0 100 16.7 0 0 1 0 0 0

D. Italy

Large mammals 0 0 0 0 0 0 0 0 0

Rodents 0 0 0 0 0 0 0 0 0

Birds 0 0 0 0 100 0 0 0 79.3

Unknown 0 0 0 0 0 0 0 0 20.7

Table 1. Relative association (%) of tick-borne pathogens with categories of vertebrate hosts determinedby blood meal analysis of unfed nymphal Ixodes ricinus.



Rickettsia helvetica and a relapsing fever-likeBorrelia sp. (RFLB) were found for the first time inIreland (0.1 and 1.1%, ticks respectively).Additionally, small numbers of ticks were infectedwith Babesia microti, B. divergens and Anaplasmaphagocytophilum (0.5, 0.1, 0.1% ticks, respectively).Although host DNA was detected in only 49.4% ofanalysed ticks, the results (Table 1A) support theview that B. garinii and B. valaisiana predominant-ly utilise song birds as reservoir hosts. It was sur-prising, however, to find that the same is evident-ly true of B. burgdorferi s.s, and that even therodent-associated B. afzelii was occasionally foundin ticks that had fed on birds.

Germany - This site is within the city limits ofBerlin and the level of pathogen-infected ticks herewas very high (55%) The majority carried B. afzelii(42% infected ticks) and the other B. burgdorferigenospecies were relatively uncommon. However,ticks infected with Rickettsia helvetica were surpris-ingly frequent (25% infected ticks). Whilst hostDNA could be detected in only 30% of analysedticks, the results suggested that R. helvetica is asso-ciated with rodents, as is B. afzelii, although, as inthe Irish study, this latter pathogen was also foundin bird-feeding ticks (Table 1B).

Spain - A total of 61 nymphal I. ricinus nymphs,collected in Spain from the Rioja area, wereanalysed. Despite the use of an additional genetarget for host DNA (12SrRNA), unambiguousdata on pathogen and/or host DNA could only beobtained for 25 specimens. Thirteen ticks wereinfected with pathogens, primarily Borreliaburgdorferi s.l., of which six contained B. afzelii, fivecontained B. valaisiana and three contained B.garinii. One tick was infected with B. microti.Detected hosts included wood mice, wild boar, reddeer, song birds and game birds. Once again thesupposedly rodent-associated B. afzelii occurred inticks that had fed on birds (game birds in this case)and there was also one positive tick associatedwith wild boar (Table 1C).

Italy - The Italian site used for tick collection islocated in central Tuscany and consists of decidu-ous woods, predominantly oak and chestnut. Noquantitative data on hosts were obtained, but deerare known to be present and song birds are abun-dant. A sample of 64 ticks was subjected to DNA

extraction in the Italian laboratory and the lysatesexamined for host DNA in Dublin. In contrast tothe situation elsewhere, a high rate of host DNAdetection was obtained (81%), but it is curious thatall of them were for birds (one game bird and 52song birds). Only 4 specimens were positive forpathogens, all of them B. garinii (Table 1D).

Discussion and conclusionsThe contrast in prevalence of tick-bornepathogens between the four countries, despitesimilarities in habitat, is striking. Whereas thebird-associated B. garinii and B. valaisiana werepredominant in Ireland, B. afzelii was the mostabundant pathogen in Germany. The small num-bers of infected ticks obtained in Spain and Italymake comparisons difficult, but it would appearthat in Italy at least, birds play a major role as tickhosts and probably import most of the ticks intothe area from locally infested regions. The lowdetection rate of host DNA in field-collected tickswas surprising since laboratory data suggestedthat 100% detection is achievable as long as 7months after the moult [1]. However, the datagenerally supported the bird-association of B.garinii and B. valaisiana, and the rodent-associa-tion of B. afzelii. It was also shown that B. afzeliimay be associated to a minor extent with birdsand wild boar, though it is possible that co-feed-ing (simultaneous feeding of infected and unin-fected ticks) or transovarial transmission (passageof pathogens via the ovary of adult female ticks tothe offspring) have a role in this apparent break-down of host specificity. Few ticks were infectedwith Anaplasma phagocytophilum and they wereinvariably associated with ruminants, as expect-ed. The pathogenic significance of the newly dis-covered Rickettsia helvetica and relapsing fever-like Borrelia sp. in Ireland and Germany requiresfurther investigation.

AcknowledgementsWe are grateful to Damian Egan (ERM) and MarkRogers (Department of Zoology, UCD) for theirassistance and to the Wellcome Trust for financialsupport.

ReferencesPichon B, Egan D, Rogers M and Gray JS, 2003. Detection

and identification of pathogens and host DNA in unfedhost-seeking Ixodes ricinus L. (Acari: Ixodidae). Journal ofMedical Entomology 40, 723–731.



Emissions of nitrous oxide from Irish soils – fertiliserand soil type effects

B. Hyde, A. Fanninga, M. Ryana, M. Hawkinsb, O. T. Cartona and P. O’Toolea Teagasc, Johnstown Castle, Co. Wexfordb Department of Statistics, UCD

Agriculture accounts for approximately 32% of greenhouse gas (GHG) emissions inIreland; emission of nitrous oxide (N2O) from soils accounts for 28% of the total agri-cultural GHG emission and is the second largest source, after methane emissionsfrom enteric fermentation in ruminants [1]. The aims of the present field- andlysimeter-scale experiments (now nearing completion) are to quantify the effects offertiliser nitrogen (N), soil type and grassland management on N2O emissions. Theresults will be compared with the Intergovernmental Panel on Climate Change(IPCC) default [N2O] emission factors [2] which are to be used in the absence ofnational [Irish] experimental data for calculating GHG emissions. The potential forchanging the IPCC default values will be evaluated in terms of their implications forIrish GHG emission inventory calculations.

Materials and methodsField experimentThe objective of this experiment is to measure N2Oemissions from grazed pasture fertilised with 0,225 and 390 kg N ha-1 corresponding to a basal orcontrol level, an application rate for an intermedi-ately stocked (1.0 Livestock units ha-1; ~170 kgorganic N ha-1) and a maximum rate forintensively stocked (2.5 Livestock units ha-1; ~250kg organic N ha-1) grassland, respectively [3]. Thesite (located at Teagasc Research Centre,Johnstown Castle, Co. Wexford) is under apermanent (>10 years old) perennial ryegrass(Lolium perenne) sward. The soil is predominantlyan imperfectly drained gley of loam over clayloam texture derived from Irish Sea till. Thegrassland management is based on rotationalgrazing (~21 days) using steers (300–350kg liveweight at spring turnout to grazing). N fertiliserapplications to the treatments throughout thegrazing season occur at the same time.

N2O emissions are measured using the closedchamber technique [4]. The chambers (11.3cm indiameter and 15.0cm high) are inserted into thesoil to a depth of 2cm to provide a seal from theatmosphere and provide an effective volumeabove the soil of 1300cm3. The chambers are left inplace for 1 hour between 10.30 and 11.30 am on

each measurement date. Samples are thenwithdrawn from the chambers using a 10mlsyringe through a rubber septum and stored inpre-evacuated 7ml screw cap septum vials for GasChromatography analysis within 6 hours. Eachmeasurement day comprises 8 chambers for eachof the 3 replicate plots per N treatment (72chambers in total).

Lysimeter experimentThe objective of this experiment is to measure N2Oemissions from grassed lysimeter units (0.6mdiameter by 1m deep) with five soil typesreceiving N fertiliser application rates similar tothose used in the field experiment. The five soiltypes represent a range of drainage classes fromseriously impeded, through moderately well-drained to very well-drained soils (Table 1). Therange is considered to be representative of themajority of Irish soils. There are nine replicatemonoliths of each soil type with three replicatesper fertiliser N treatment on each soil type. Thegrass (L. perenne) in the lysimeter unit is cut at six-week intervals from March through to November.Nitrogen is applied in four to five equal applica-tions during the growing season. A single N2Osampling chamber, as described above, is used oneach lysimeter column.



Preliminary resultsField experiment Two preliminary models of N2O emissions fromgrazed grassland have been developed: (i) amultiple regression model based on average dailyemissions per replicate plot in which the importanceof causative factors, the nature of their influence, andtheir interrelationship can be identified; and (ii) amixture model in which individual flux chamber dataare modelled (i.e. the raw data are modelled).

Lysimeter experimentThe preliminary results are in line with expectedtrends. The soil effect on N2O emissions appears to beas great as the fertiliser effect with higher emissionsassociated with the seriously impeded soils (Figure 1).

ConclusionsIn both experiments, larger N2O emissions areassociated with higher N fertiliser inputs, wet

rather than dry conditions and in the periodsimmediately following the fertiliser applications.

AcknowledgementsThis work is funded by Teagasc and the EPA underthe ERTDI programme as part of the Irish NationalDevelopment Plan 2000–2006.

References1. Anon, 2000. National climate change strategy 2000. Department

of Environment, Heritage and Local Government,Stationary Office, Dublin.

2. IPCC, 1997. Revised 1996 guidelines for national greenhouse gasinventories: Reference Manual, Chapter 4. IntergovernmentalPanel on Climate Change, OECD, Paris.

3. Coulter BS (ed), 2001. Nutrient and trace element advice forgrassland and tillage crops. Teagasc, Johnstown CastleResearch Centre, Wexford.

4. Smith KA, Clayton H, McTaggart IP, Thomson PE,Arah JRM and Scott A, 1995. The measurement ofnitrous oxide emissions from soil using chambers.Philosophical Transactions of the Royal Society, London351, 327–338.

Castlecomer Poorly drained organic clay loam over clay loam texture

Rathangan Poorly drained loam overlying clay loam structure

Elton Well drained gravelly loam overlying gravelly clay loam texture

Clonroche Well drained loam to clay loam, overlying shaley loam textured brown earth

Oakpark Gravelly brown earth of course sandy loam texture

Table 1. Experimental soil types used in the lysimeter unit.

Fig. 1. Effect of soil type and N application rate on N2O emissions.



The impact of forestry operations on phosphorus insurface waters at an ex-agricultural siteO. M. McCabe, E. P. Farrell and P. O’Dea1

1Coillte Teoranta Research Laboratory, Newtownmountkennedy, Co. Wicklow

In the PEnrich project, the impacts of current forestry practices on water quality areexamined, with particular emphasis on phosphorus. The efficacy of the Forestry andWater Quality Guidelines in minimising phosphorus loss will be assessed andwhere necessary recommendations will be made for improvements to these guide-lines. The impacts of alternative silvicultural techniques (continuous cover forestryand group felling) on water quality will also be examined.

IntroductionIn recent decades there has been much concernabout phosphorus (P) loss from land to water andeutrophication of Irish rivers and lakes. Forestryactivities have been identified as one of thepotential risks to water quality and have beenexamined in a number of studies carried out inIreland [1,2,3]. The impact of forestry operationson water quality has also been examined withinrecent catchment-based studies at Lough Leaneand the Three Rivers Boyne, Liffey and Suir [4, 5].

Over the past fifteen years there has been a moveaway from poor mineral upland and peat soilstowards afforestation of land previously used foragricultural purposes. Soils in these areas can bequite different to those that traditionally underwentafforestation and little is known about the impact offorest establishment on water quality at such sites –an issue to be addressed in the PEnrich project(Forestry Operations and Eutrophication).

The main aims of the PEnrich project are to monitorthe impact of current forestry practices on phosphorusin surface waters and to assess the efficacy of Forestryand Water Quality guidelines in minimisingphosphate loss. Recommendations for amendmentsand/or improvements to these guidelines and to theCode of Best Forest Practice will be considered on thebasis of the findings of the project. In this report,preliminary findings are presented of a study carriedout at an ex-agricultural site in Co. Mayo.

Materials and methodsThe study site (Figure 1) is located nearCrossmolina, Co. Mayo and is approximately1.5km from the shores of Lough Conn. The site

Fig. 1. Ex-agricultural study site, Co. Mayo, pre- and post-forestry operations.



covers an area of 3.5ha and is isolated by water-courses on all sides. It received annual applica-tions of NPK fertilizer until 2000. It is a poorlydrained soil with a Morgan’s P content of 14.2 to19.1mg l-1 in the upper layer (0–5cm)

Sampling began in June 2002, nine months priorto any forestry operations, thus enabling us toestablish baseline data of water quality at the site.The sampling program is intensive, consisting of:(i) weekly grab samples from various subsurfacedrains, (ii) composite weekly samples at each ofthe drains/streams leading into the main drainageoutlet, and (iii) composite daily samples (collectedon an hourly basis) from the main drainage outletof the site using a Sigma 900 Max autosampler.

Samples are analysed for P fractions (totalreactive P [analysed since December 2003],dissolved reactive P and total P), cations (Mg, Ca,Fe, K, Na, Mn and Al), anions (SO4

2-, NO3- and Cl-

), suspended solids, pH, conductivity, alkalinityand ammonia in the laboratories at UCD andCoillte. Flow readings for the main drainage outletof the site are also recorded to enable us toquantify nutrient loading. In this paper, resultswill be discussed in terms of total reactive P con-centrations (TRP) only, i.e. molybdate reactive P inunfiltered water samples.

Forestry operations at this site commenced inMarch 2003. This involved the installation ofmound drains followed by planting a mixture ofoak and Scots pine. Sampling will continue at thissite until August 2004.

Results and discussionHigh total reactive P (TRP) concentrations wereobserved for water samples collected fromsubsurface drains and streams/drains leading intothe main drainage outlet of the site on manyoccasions prior to and after forestry operations(Table 1). TRP values often exceeded the 30µg l-1

threshold outlined in the Phosphorus Regulations[6] as the target for good water quality. This may berelated to higher suspended sediment concentra-tions also observed on these occasions. While high

TRP concentrations have been detected on manyoccasions since forestry activities began, thesevalues do not appear to differ significantly fromTRP concentrations measured before this period.

In general, TRP concentrations of samplescollected from the main drainage outlet of the sitewere relatively low before and after forestryoperations. TRP concentrations in the main drainageoutlet ranged between 7.6 and 32.8µg l-1 prior to anyforestry operations (Table 1). During this period,TRP concentrations exceeded the 30µgl-1 thresholdon only two occasions. This recommended targetvalue has been exceeded on four occasions in theperiod 19 March to 31 December 2003. The mostsignificant of these events was on 12 April when aTRP concentration of 334.2µg l-1 was observed.There is no indication that this high TRP concentra-tion is related to any increase in sediment loss at thetime. TRP values observed in subsequent days wereless than 30µg l-1. The significance of this event willbe further investigated.

It seems that while TRP concentrations instreams and drains leading into the main drainageoutlet of this site are often quite high, these valuesare diluted substantially as the water moves intothe larger stream. Thus, TRP concentrations insurface water leaving this site are generally low.

Early indications are that with the exception ofone occasion, forestry operations at this site have nothad any major adverse impacts on water quality.Other nutrient concentrations and loads have yet tobe examined and this will give us a better under-standing of water quality as a whole at this site.

AcknowledgementsThe authors wish to acknowledge COFORD andEPA for funding this research. We also wish tothank Erica Geurts, Norman Butler, MarinaConway (UCD and FERG) and staff at the Coillteresearch lab for their assistance.

Further information on the PEnrich project can befound at the following website:http://www.ucd.ie/ferg/Research/Projects/Penrich

Date Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Range 7.6-121.2 3.7-24.52 2.8-33.0 0.7-334.2 3.8-27.6 1.5-45.0 nd-31.7 nd-11.2 1.2-16.0 1.0-20.9 7.3-32.7 9.8-22.1

mean 22.6 11.9 8.1 20.8 9.4 8.4 10.1 3.5 4.8 6.7 16.1 14.3

SD 30.4 6.0 6.5 66.9 4.9 10.6 5.9 3.2 3.5 5.5 8.8 5.4

Table 1. Range, mean and standard deviation (SD) of TRP concentrations (µg l-1) in the main drainageoutlet of the study site during 2003; nd=not detectable.



Ground vegetation management on afforested cutawaypeatlands in the Irish midlandsF. Renou and E. P. Farrell

Cutaway peatlands are areas left over after peat harvesting. In Ireland, as much as50,000 hectares of these peatlands have the potential to become an important sitetype for forestry development. The BOGFOR research programme was developed inorder to study optimum means of growing various tree species to their full potentialwithout compromising the environment. While competition from ground vegetationcan be a serious barrier to adequate tree growth, the conventional management ofweeds in these new forests is restricted and new methods such as mowing and weedwiping have been shown to be efficient.

As a result of peat harvesting for horticulture orenergy generation, large areas of bare cutawaypeatlands are left for other land uses. Naturalrecolonisation can be slow due to the harsh environ-mental conditions. Depending on the surroundingseed source, birch, ericaceous shrubs and rushes areoften the only species found even 10 years afterharvesting has ceased. The flora is somewhatboosted when fertiliser is applied after tree planting.Competition from ground vegetation can have anadverse effect on the growth and survival of the treecrop. However, conventional vegetationmanagement methods are not suited to localconditions. Information on the level of competitionencountered has been gathered and newmanagement practices have been tested.

Materials and methodsThis study is part of a larger investigation into thevegetation ecology of afforested cutaway peatlands.Three vegetation surveys were carried out in 1999

[1], 2001 [2] and 2002 [3] on over one hundredquadrats (10m × 10m each) covering 15 differentsites in county Offaly. It was found that several envi-ronmental factors affect the level of competitionbetween ground vegetation and planted trees,including the age of the cutaway, the pH of the peatand water table levels. In all cases however, somedegree of weed control was required for satisfacto-ry tree growth. Observations were recorded fromregularly managed sites and permanent quadratswere selected to test new vegetation managementtreatments: mowing and/or weed wiping. Thevegetation response to the treatments is beingmonitored and will continue to be until such time asthe trees are tall enough to shade the weeds.

Results and discussionWeeds can be as great a problem on peat than onmineral soils. Soon after fertiliser is applied atplanting time, weeds quickly cover the ground,but they normally consist of fewer species in

References1. Renou F, Jones SM and Farrell EP, 2000. Leaching of

phosphorus fertiliser applied on cutaway peatland forestsrecently established in central Ireland. Sustaining ourPeatlands: Proceedings of the 11th International Peat Congress,Québec, Canada, Vol II, 984–990.

2. Cummins T and Farrell EP, 2000. Effects of forest operationson acid peatland streamwater composition. Proceedings ofthe International Association of Theoretical and AppliedLimnology 27, 1556–1559.

3. Cummins T and Farrell EP, 2003. Biogeochemical impacts ofclearfelling and reforestation on blanket peatland streamsI. phosphorus. Forest Ecology and Management 180, 545–555.

4. Kirk McClure Morton and Pettit Project Design andManagement, 2000. A catchment based approach forreducing nutrient inputs from all sources to the Lakes ofKillarney: Second interim report. Lough Leane Catchment,Monitoring and Management System, Kerry CountyCouncil, Tralee.

5. Earle JR, 2003. The Three Rivers Project – water qualitymonitoring and management systems in the Boyne, Liffeyand Suir catchments in Ireland. Water Science andTechnology 47, 217–225.

6. Anon, 1998. Statutory Instrument No. 258 of 1998. LocalGovernment (Water Pollution) Act, 1977 (Water QualityStandards for Phosphorus) Regulations, 1998.



higher numbers. Also, more weed flushes occurduring the season and weed seedlings can emergefrom a greater depth than on mineral soil. Theeffect of cultivation such as deep ploughing hasbeen found to reduce weed populations in the firstyear after scarification. Elsewhere, peat that hasbeen mixed with mineral soil presents evengreater problems than pure peat soils in terms ofweed diversity and competition.

While common bent (Agrostis capillaris), rosebaywillowherb (Chamaenerion angustifolium), downybirch (Betula pubescens) and willow species (Salixspp.) are the most common species on driercutaway peatlands, soft rush (Juncus effusus) is themost dominant species. This poses problems as itcompetes strongly with the trees [4].

Unfortunately, neither residual nor foliarherbicides can be used on these particular sites,because the former are strongly adsorbed onorganic matter and the latter should not be appliedto wet foliage, when rain is imminent or whenthere is a danger of drift. In open, windy peatlandareas there are usually few days in the earlygrowing season when ideal spraying conditionsoccur.

Mowing has been carried out for severalseasons at some sites where a small all-terrainvehicle has been used to tow a purpose-built flailbetween the rows of trees (Figure 1). Evidencesuggests that the timing of mowing is important. Ifthere is to be only one mowing, the optimum timeis in mid-summer when growth rates are highest.However, other observations indicate that forgood control, repeated treatments (2–4) arenecessary, especially when the site is colonisedwith rushes. On several sites, soft rush (Juncuseffusus) has ‘swamped’ the trees and could only bemowed at half their height, which had nonoticeable effect.

A second method was tested using a weed-wiper (Figure 2). It consisted of a carpeted rollerwetted with a contact herbicide (Glyphosate) by amanually controlled 12V pump. The vegetation iswiped twice, the second time in reverse direction.While the rushes died off in most plots, vegetationcover did not decrease as much as after mowing.In the context of reducing the use of herbicides innew Irish forests, weed wiping should only beused in cases where mowing is not feasible.

Finally, on waterlogged sites, it may be sensible toretain some natural vegetation such as rushes toact as a water consumer.

AcknowledgementsThis work was funded by Bord na Móna, Coillteand COFORD (National Development Plan).

References1. Parr SL, 1999. Baseline studies on ground vegetation in

plantation forests in cutaway bog. Forest EcosystemResearch Group Report 65, Department of EnvironmentalResource Management, UCD, 53 pp.

2. Pöllänen M and Renou F, 2002. Development of plantationforest on industrial cutaway peatlands in relation toground vegetation – 2001 survey. Forest Ecosystem ResearchGroup Report 66, Department of Environmental ResourceManagement, UCD, 50 pp.

3. Bennett C, Pöllänen M and Renou F, 2003. Vegetation dynamicson afforested cutaway peatlands in the Irish midlands. ForestEcosystem Research Group Report 68, Department ofEnvironmental Resource Management, UCD, 54 pp.

4. McCorry MJ and Renou F, 2003. Ecology and managementof Juncus effusus on cutaway peatlands in the Irishmidlands. Forest Ecosystem Research Group Report 69,Department of Environmental Resource Management,UCD, 75 pp.

Fig. 1. Flail mower.

Fig. 2. Roto-wiper behind a quad bike.



Inter-annual and seasonal variation of methaneemissions in a wetland plant communityD. Wilson, J. Alma, J. Laineb, K. A. Byrnec and E. P. Farrella Finnish Forest Research Institute, Finlandb University of Helsinki, Finlandc Department of Civil and Environmental Engineering, University College Cork

The role of wetland plant communities in regulating methane emissions is wellknown. However, emissions are subject to considerable variability both spatiallyand temporally. The aim of this study was to determine the inter-annual andseasonal variation in methane emissions in one of the most widely distributed plantcommunities in restored cutaway peatlands in Ireland.

Methane (CH4) is an important greenhouse gas andits concentration in the atmosphere has doubledover the last two hundred years. Natural wetlandsrelease around 115 Tg (1Tg = 1012g) of methane peryear [1], primarily as a result of the anoxicconditions in the soil profile caused by a persistent-ly high water table. The role of vegetation inregulating CH4 fluxes is also of great importance[2,3]. Wetland plants provide a medium for methaneproduction through the release of labile compoundsin root exudates and litter input, and they alsofacilitate the movement of CH4 from anoxic peat tothe atmosphere through specialised internal cellstructures. Given that there could be up to 30,000hectares of cutaway peatland available forrestoration to wetlands in Ireland over the next30–40 years, there is an obvious need to quantify thepotential methane emissions from these newecosystems.

Materials and methodsThe study was carried out on a cutaway peatland atTurraun, Co. Offaly, as part of a larger study investi-gating carbon gas fluxes within a range of wetlandvegetation communities. Following the cessation ofindustrial harvesting in the 1970s, the drainageditches were blocked in 1990 and the cutaway wasreflooded to encourage the development of awetland ecosystem. Since that time, a diversepattern of vegetation communities has becomeestablished on the cutaway. At the beginning of thestudy, four permanent stainless steel collars (60cmby 60cm) were established within an Eriophorumangustifolium (bog cotton) / Carex rostrata (bottle

sedge) plant community in order to monitor,measure and assess the contribution of thatcommunity to CH4 gas emissions over a 2-yearperiod. Wooden boardwalks were constructed toprevent damage to the vegetation and peat surface.CH4 measurements were carried out at 1–2 weekintervals from June to December in 2002 and 2003.Gas samples were collected using a static chambermethod [4] (Figure 1).

Four 40ml samples were withdrawn into 60mlpolypropylene syringes from the chamberheadspace at 5-minute intervals. At the same time,air temperature inside the chamber, soiltemperature at 2, 5, 10, 20 and 30cm depths andwater table depth were recorded. Gas sampleswere analysed for CH4 concentration within 24hours of collection with a gas chromatograph(Shimadzu GC-14-B) using a flame ionisationdetector (FID). CH4 fluxes (mg m-2 h-1) werecalculated from the linear change in CH4 concen-

Fig. 1. Static chamber system used for methane (CH4)sampling in Turraun, Co. Offaly.



tration in the chamber headspace over time.Positive flux values indicate movement of CH4

from the soil to the atmosphere. Negative valuesindicate uptake of CH4 by the soil.

Results and discussionThe mean CH4 flux rates differed significantlybetween the summer months of 2002 and 2003(P<0.001) with maximum mean flux rates of1.61mg m-2 h-1 and 0.7mg m-2 h-1 observed in Juneand August respectively (Figure 2). In both years,CH4 fluxes decreased during the autumn with theonset of senescence in the vegetation. However,there was no significant difference in the amountof CH4 released between the winter months of2002 and 2003.

Temporal variation in CH4 fluxes, both seasonaland inter-annual, have been reported in othersstudies [5]. The production, transport andemission of CH4 are controlled by a large numberof variables such as temperature, water tableposition, nutrient status of the soil, microbialpopulation and the plant species themselves.These variables can change from year to year witha subsequent effect on methane dynamics.

ConclusionsResults from this study suggest that Eriophorumangustifolium / Carex rostrata plant communities arelikely to be sources of CH4 but the magnitude ofthe emissions may vary seasonally and inter-annually depending on the prevailing environ-mental conditions.

AcknowledgementsThis research was funded by Bord na Móna.

References1. Matthews E and Fung I, 1987. Methane emission from

natural wetlands: global distribution, area and environ-mental characteristics of sources. Global BiogeochemicalCycles 1, 61–86.

2. Sebacher DI, Harriss RC and Bartlett KB, 1985. Methaneemissions to the atmosphere through aquatic plants.Journal of Environmental Quality 14, 40–46.

3. Frenzel P and Rudolph J, 1998. Methane emission from awetland plant: the role of CH4 oxidation in Eriophorum.Plant and Soil 202, 27–32.

4. Crill PM, 1991. Seasonal patterns of methane uptake andcarbon dioxide release by a temperate woodland soil.Global Biogeochemical Cycles 5, 319–334.

5. Tuittila E-V, Komulainen V-M, Vasander H, Nykanen H,Martikainen PJ and Laine J, 2000. Methane dynamics of arestored cut-away peatland. Global Change Biology 6,569–581.

Fig. 2. Mean flux rates of CH4 (mg m-2 h-1) + standard error at sample site from June to December 2002 and 2003.



Mapping the geography of the postglaciallandscapeJohn Feehan and Louise Dunne

Since the astonishing discovery of a Mesolithic camp carbon-dated to around 9,200 y.a.under the deep raised bog at Boora in Co. Offaly in the 1970s, interest in the geographyof the pre-bog landscape has been growing among archaeologists [1]. At the end of theIce Age 10,000 or so years ago, before the raised bogs extended their hold over centralIreland, the water table over much of the country was several metres higher than itspresent level. A key piece of evidence for the limits of the lakes that dominated thelowlands at this time survives in the form of limestone boulders and bedrock eroded bywave action at the edges of early postglacial lakes which have since vanished or retreat-ed. These solitary sentinels still bear the unmistakable marks of long-continued erosionby lapping waves at the lake edge. They are sometimes shaped like mushrooms; othershave an overhang facing in one direction, but all are notched and undercut in a fashionas to suggest prolonged exposure to standing water at some time in the past. Themushroom-shaped stones are produced where the notching forms a fairly even circlearound the stone, the flat underside of the mushroom marking the former lake-level.The wavestones are always of limestone: generally isolated boulders which are oftenglacial erratics, or (more rarely) exposed bedrock (Figure 1).

By recording these features on a map we are ableto plot the original shoreline of the earlywaterworld in which the first Irish people lived.These wave-moulded stones are most frequent inthe Shannon basin, especially on the western shoreof Lough Ree and between Clonmacnois andShannonbridge, where their location and the lakeheight recorded by their characteristic lip helps usto plot the extent of the postglacial lake network inthis area. Wave-worn boulders are also found here

and there around the Midland bogs, often milesfrom any modern lake; these are even moresignificant milestones marking the position of theancient and now vanished pre-bog shoreline(Figure 2). This is highly significant in archaeolog-ical terms because this shoreline was the focus ofsettlement for the earliest human communities inthis part of Ireland. Tracing that shorelinetherefore helps us to pinpoint the areas wheresettlement evidence is most likely to be found.

Fig. 1. This stone at Cornaseer, west of Ballybay in Co.Roscommon, can be regarded as a sort of ‘type specimen’ forall mushroom stones. It is the most striking of a large group ofwave-eroded boulders found along the western shore of LoughRee. It has the classic mushroom form, and exhibits perfectlythe double lip – recording a two-stage drop in lake level – foundon many of the mushroom stones in the midlands.

Fig. 2. The Crancreagh stone near the edge of Derrinlough bogin Co. Offaly.



Wavestones are also known from other parts ofIreland. They occur in the Corrib floodplain. InClare they have been found especially along theFergus and at the foot of Mullaghmore (Figure 3),recording a substantially higher water table at someas yet undefined time in the past. But if the watertable was higher where these marker stones occur, itwas higher everywhere in the Burren lowlands. Thismakes for an altogether different land-waterinterface from that which we see today: and that hasimplications for what the resource base of earlypeoples and their settlement patterns were.

A recent survey has plotted the location of allknown mushroom stones, using GPS to measure thevanished water levels they record [2,3]. This figurecan be extrapolated onto a local contour map to plotthe run of the ancient shoreline. Several new stonescame to light during the course of this study, andthere can be little doubt that many others awaitdiscovery. To heighten public awareness of thenature and importance of these fragile features anattractive full-colour booklet has been producedwith the assistance of the Heritage Council [4](Figure 4), and a website set up to record and reportfurther finds, and to promote further study:http://www.ucd.ie/envinst/envstud/mushroomstones

References1. Feehan J and O’Donovan G (1996). The Bogs of Ireland, an

Introduction to the Natural, Cultural and Industrial Heritage ofIrish Peatlands. UCD, The Environmental Institute, 518 pp.

2. Dunne LA, 1998. The cataloguing, mapping, photographing andmeasuring of the lip height of ‘mushroom stones’ with an inves-tigation into the possible factors involved in their formation andconservation recommendations. Unpublished M.Sc. thesis,ERM.

3. Dunne LA and Feehan J (2003). The origin and significanceof mushroom stones in lowland karst regions. Irish Journalof Earth Sciences, 20: 33–40.

4. Dunne, L. and Feehan, J. (2003). Ireland’s Mushroom Stones:Relics of a Vanished Lakeland. UCD, Department ofEnvironmental Resource Management.

Fig. 3. The Crossard stone in the Fergus floodplain at the edgeof the Burren.

Fig. 4. The cover of the mushroom stones booklet. This is the second in a series of occasional lanscape studies published by theDepartment of Environmental Resource Management.



Detection and quantification of soil-borneinoculum of Spongospora subterranea f. sp.subterranea, the cause of powdery scab disease ofpotatoJ. M. Tuohy, X. Qu, D. Egan, J. A. Kavanagh and F. M. Doohan

In this investigation a competitive PCR assay for the quantification of Spongosporasubterranea f. sp. subterranea, the cause of powdery scab of potato, in soils ofdifferent texture and naturally infested field soils was developed. Heavily infestedsoils which carry a high risk of powdery scab can be identified before planting andgrowers can now avoid serious reduction in tuber quality and yield losses. Thismakes an important contribution to the development of a management programmefor this disease, since no highly resistant cultivars or environmentally acceptablechemicals are currently available for disease control.

IntroductionSpongospora subterranea f. sp. subterranea is a soil-borne obligate parasite that causes powdery scabdisease of potato and survives for many years in soilas cystosori (spore balls) consisting of aggregates ofresting spores [1]. Under favourable conditionscystosori germinate and each resting spore releasesa single zoospore that infects roots and stolons [2].There is little information on the levels of cystosoriinoculum in field soils but it has been suggested thatsoils heavily infested with S. subterranea containseveral hundred cystosori per gram of soil [3]. In thedevelopment of a management strategy forpowdery scab accurate determination of cystosorinumbers in soils is important in identifying fieldsoils at risk to this disease.

Molecular methods based on the polymerasechain reaction (PCR) [4,5] have been developedfor the detection and quantification of S.subterranea cystosori in spiked soils in thelaboratory and more recently cystosori andzoospore inoculum levels in soil, water andtubers have been assayed using real time PCR [6].The objective of this study was to refine anddevelop a quantitative PCR-based method [5] forthe detection and quantification of S. subterraneacystosori in soils of different texture and toconsider the feasibility of its application for theroutine testing of soils as part of a risk assessmentstrategy for powdery scab disease.

Materials and methodsNon-infested (Spongospora-free) soil samples,collected from three sites with no previous potatocropping history, were air-dried and sieved(2mm). Soil texture analysis was performed usinga standard method [7]. Soil sub-samples (0.25g)(further sieved through a 100 µm mesh) werespiked with aliquots (100 µl) of serial dilutions ofcystosori (5-103 per gram soil) prepared frompowdery scab lesions. DNA was extracted fromeach soil sub-sample using an UltraClean SoilDNA Kit. Species specific PCR was performed toconfirm the presence of S. subterranea DNA in soilDNA extracts [5]. Quantitative PCR analysis wascarried out using a heterologous competitortemplate (1pg per reaction) with flanking S.subterranea specific primer sites (SBITSA, SBITSD)(5). Quantitative PCR was conducted as describedby Qu et al. [5], except that the programmeconsisted of 30 PCR cycles. Standard curves wereconstructed by plotting PCR co-amplified S.subterranea target sequence (434 bp) to competitortemplate (541 bp) ratios obtained for each soil typeand the mean obtained for all soil samples againstthe cystosori content of spiked soil.

This method was then used for the quantifica-tion of cystosori in thirteen naturally infested fieldsoils and two garden soils from different locationsby subjecting 1µl soil DNA extracts for PCRanalysis (as above). The resulting target to



competitor ratios were used to extrapolate thecystosori content from each of the quantitativePCR standard curves. All quantitative PCR resultswere based on at least two replicate PCR reactionsof two replicate DNA extractions for each soilsample. Data were analysed using the nonpara-metric Kruskal-Wallis Test in StatView 5.0.

ResultsThe three Spongospora – free soils used for thegeneration of standard curves (5-103 cystosoriper gram soil) were classified as clay loam andsandy-clay-loam (Table 1). Standard curvesgenerated for each soil type (Figure 1) regressedthe S. subterranea target (434 bp) to heterologouscompetitor (541 bp) product ratios against thelog number of spiked cystosori. The meanregression coefficient of r2 = 0.99 (Figure 1)indicates that PCR quantification of S.subterranea cystosori in soil of differing texturewas not only accurate but also reproducible.Statistical analysis showed that there were nosignificant differences between the PCR productratios obtained for the three soils at any concen-tration of cystosori tested (P≥ 0.05).

S. subterranea cystosori in thirteen infested fieldsoil samples (S1-S13) and two garden soils (G1,G2) were then detected and quantified using thisPCR assay. All thirteen field soils were infestedwith S. subterranea cystosori but the garden soilswere not. The numbers of cystosori per gram soilin each soil were extrapolated from equations

produced from each soil type and the overall mean(Table 2). Cystosori were successfully quantifiedin field soils. Results showed that soils containedbetween 1,134 and 55,648 cystosori per gram ofsoil. Also, no significant differences were foundfor any soil in the cystosori number calculatedusing either the different standard curves or thestandard curve of the mean of the three soils.

DiscussionIn this study it was confirmed that the S.subterranea – specific PCR primers developed inthis laboratory [5] can detect this pathogen in soilsof different textures. The primers were also suc-cessfully used for the quantification of cystosori ina range of field soils naturally infested with S.subterranea and non-infested garden soils fromdifferent locations. This quantitative PCR assay isnot only sensitive and specific but also veryaccurate and reproducible, detecting approximate-ly 5 cystosori per gram of soil. It should be notedthat the PCR method for the detection and quan-tification of S. subterranea in tubers and soil [4] anda similar method [6] using real time PCR for S.subterranea quantification in soil, water and tuberswhile detecting <5 cystosori per gram soil, involvea more complex DNA extraction procedureincluding bead beating and Sephadex purificationthat takes approximately four hours. TheUltraClean TM Soil DNA extraction protocol usedin this investigation only takes only one hour andfewer procedural steps. Real time PCR however,

�

�

�

��

Fig. 1. Spongospora subterranea quantitative PCR standard curves, relating the S. subterranea - specific target to competitor PCRproduct ratios to the number of cystosori in three different soils. The mean standard curve for the three soil types is also shown.



though requiring more costly equipment, is morerapid than the conventional quantitative PCRused here. It should be pointed out also that theassay described by Bell et al. [4] was not used forthe quantification of naturally infested field soilsor soils of different textures and that the detectionand quantification of cystosori in clay soil was notconsistent, possibly due to DNA binding or PCRinhibition [6]. It is concluded that the quantitativePCR method used in this investigation is veryadaptable and can be recommended withconfidence for the routine quantification of S.

subterranea in field soils as part of a riskassessment management strategy for powderyscab disease.

AcknowledgementsThis work was funded by the NationalDevelopment Plan (NDP) Research Stimulus Fund(Department of Agriculture, Food and RuralDevelopment) (J. M. Tuohy) and in part by aTeagasc Walsh Fellowships (X. Qu) and theNational Veterinary and Biotechnology Centre(UCD).

Soil Origin of sample % Sand % Silt % Clay Texture

1 Co. Westmeath 29.9 36.3 33.8 Clay-loam

2 Co. Kildare 47.5 20.0 32.5 Sandy-clay loam

3 Co. Dublin 34.7 27.9 37.5 Clay-loam

Table 1. Origin and texture analysis of three Irish soil samples.

Table 2. Number of cystosori g-1 soil for fifteen field and garden soil samples, as determined by quanti-tative PCR analysis of soil DNA extracts and extrapolated through equations for each soil type or meanequation from standard curves.

a Soils prefixed by G originated from gardens; those prefixed by S originated from field sites. b Mean cystosori pergram soil ± standard error for any given sample; values extrapolated from each standard curve were not signifi-cantly different (P≥ 0.05).

Cystosori (g-1 soil) b

Soil sample a Standard curveSoil 1

Standard curveSoil 2

Standard curveSoil 3

Mean standardcurve (3 soils)

G1 0 ± 0 0 ± 0 0 ± 0 0 ± 0

G2 0 ± 0 0 ± 0 0 ± 0 0 ± 0

S1 3168 ± 183 3769 ± 216 3541 ± 210 3485 ± 202

S2 4805 ± 62 5691 ± 73 5434 ± 73 5299 ± 69

S3 14589 ± 174 16952 ± 199 17356 ± 212 16259 ± 194

S4 4891 ± 121 5791 ± 141 5534 ± 141 5394 ± 141

S5 9058 ± 329 10579 ± 381 10634 ± 397 10068 ± 368

S6 4823 ± 336 5754 ± 397 5351 ± 383 5297 ± 371

S7 7395 ± 334 8783 ± 393 8302 ± 386 8143 ± 370

S8 49977 ± 3451 58165 ± 3974 59200 ± 4202 55648 ± 3865

S9 6853 ± 785 7668 ± 910 7613 ± 938 7259 ± 876

S10 14233 ± 884 16541 ± 1017 16921 ± 1081 15859 ± 992

S11 1033 ± 6 1235 ± 7 1141 ± 7 1134 ± 6

S12 1934 ± 13 2295 ± 16 2174 ± 15 2129 ± 14

S13 8553 ± 1312 9993 ± 1517 10026 ± 1581 9502 ± 1466



References1. Kole AP, 1954. A contribution to the knowledge of

Spongospora subterranea (Wallr) Lagerh., the cause ofpowdery scab disease of potatoes. Tijdschrift overPlantenziekten 60, 1-65.

2. Lahert H and Kavanagh JA, 1985. The fine structure of thecystosorus of Spongospora subterranea, the cause ofpowdery scab of potato. Canadian Journal of Botany 63,2278-2282.

3. Merz U, 1993. Epidemiological aspects of powdery scabof potatoes caused by Spongospora subterranea. 2nd

Symposium of the International Working Group on PlantViruses with Fungal Vectors, Proceedings. Denver, USA:The American Society of Sugar Beet Technologists, 103-106.

4. Bell KS, Roberts J, Verrall S, Cullen DW, Williams NA,Harrison JG, Toth IK, Cooke DE, Duncan JM and ClaxtonJR, 1999. Detection and quantification of Spongosporasubterranea f. sp. subterranea in soils and on tubers usingspecific PCR primers. European Journal of Plant Pathology 105,905-915.

5. Qu X, Kavanagh JA and Egan D, 2000. Quantification of thefungal cause of potato powdery scab in soil using acompetitive polymerase chain assay. Research Report 1998-1999, Faculty of Agriculture, University College Dublin, 74-76.

6. Van der Graaf P, Lees AK, Cullen DW and Duncan JM, 2003.Detection and quantification of Spongospora subterranea insoil, water and plant tissue samples using Real Time PCR.European Journal of Plant Pathology 109, 589-597.

7. Kilmer VJ and Alexander LT, 1949. Methods of makingmechanical analyses of soils. Soil Science 68, 15-24.



Journal articlesAbosriwil S and Clancy KJ (2002). A protocol for

evaluation of the role of disinfectants in limitingpathogens and weed moulds in commercialmushroom production. Pest ManagementScience, 58: 282–289.

Aherne, J, Kelly-Quinn M and Farrell EP (2002). Asurvey of lakes in the Republic of Ireland:hydrochemical characteristics and acidsensitivity. Ambio, 31:452–459.

Aherne J and Farrell EP (2002). Deposition ofsulphur, nitrogen and acidity in precipitationover Ireland: chemistry, spatial distribution andlong-term trends. Atmospheric Environment, 36:1379–1389.

Aherne J and Curtis CJ (2003). Critical loads ofacidity for Irish lakes. Aquatic Sciences, 65: 21–35.

Ansari KI, Palacios N, Araya C, Langin T, Egan Dand Doohan FM (2002). Genetic diversityamong Colletotrichum lindemuthianum isolatesfrom Central and Latin America, Africa andEurope. Plant Protection Science, 38: 378–380.

Arnett RTP and Whelan J (2001). Comparing thediet of cod (Gadus morhua) and grey seals(Halichoerus grypus): an investigation ofsecondary ingestion. Journal of the MarineBiological Association of the United Kingdom, 81:365–366.

Brennan J, Egan D and Doohan FM (2002).Characterization and differentiation of IrishErwinia amylovora isolates. Journal ofPhytopathology, 150: 414–422.

Brennan JM, Fagan B, van Maanen A, Cooke BMand Doohan FM (2003). Studies on in vitrogrowth and pathogenicity of EuropeanFusarium fungi. European Journal of PlantPathology, 109: 577–587.

Brennan M and Whelan J (2000). A comparativestudy of the bird communities in coniferous andbroadleaved woodland at various stages in thegrowth cycle. Irish Forestry, 57: 11–17.

Browne RA, White EM and Burke JI (2002).Hullability of oat varieties and its determina-tion using a laboratory dehuller. Journal ofAgricultural Science, 138: 185–191.

Browne RA, White EM and Burke JI (2003). Effectof nitrogen, seed rate and plant growthregulator (chlormequat chloride) on the grainquality of oats (Avena sativa). Journal ofAgricultural Science, 141: 249–258.

Chauzat MP, Purvis G, and Dunne R (2002).Release and establishment of a biologicalcontrol agent, Psyllaephagus pilosus (Noyes)(Hym. Encrytidae) for eucalyptus psyllid,Ctenarytaina eucalypti (Maskell) in Ireland.Annals of Applied Biology, 141: 293–304.

Collins R (2002). Sex differences in the movementsand mortality of Mute Swans. Waterbirds, 25(Suppl. 1): 157–161.

Cummins EJ, Grace PM, McDonnell KP, Ward SMand Fry DJ (2001). Predictive modelling and riskassessment of BSE: A review. Journal of RiskResearch, 4: 251–274.

Cummins EJ, Colgan SF, Grace PM, Fry DJ,McDonnell KP and Ward SM (2002). Humanrisks from the combustion of SRM-derivedtallow in Ireland. Human and Ecological RiskAssessment, 8: 1177–1192.

Cummins E, Grace P, Fry J, McDonnell K andWard S (2002). BSE: Risk, uncertainty and policychange. Risk Health, Safety and Environment, 13:95–113.

Cummins EJ, Grace PM, Fry DJ, McDonnell KP,Colgan SF and Ward SM (2002). Quantitativeexposure assessment for the combustion ofmeat and bone meal derived from specifiedrisk material in the context of BSE in Ireland.Journal of Agricultural Safety and Health, 8:356–383.

Cummins T and Farrell EP (2003). Biogeochemicalimpacts of clearfelling and reforestation onblanket peatland streams. 1. phosphorus. ForestEcology and Management, 180: 545–555.

Cummins T and Farrell EP (2003). Biogeochemicalimpacts of clearfelling and reforestation onblanket peatland streams. 2. major ions anddissolved organic carbon. Forest Ecology andManagement, 180: 557–570.

Curry JP, Byrne D and Schmidt O (2002). Intensivecultivation can drastically reduce earthwormpopulations in arable land. European Journal ofSoil Biology, 38: 127–130.

Diamond H and Cooke BM (2002). Preliminarystudies on biological control of the Fusarium earblight complex of wheat. Crop Protection, 22:99–107.

Doohan FM, Brennan J and Cooke BM (2003).Influence of climatic factors on Fusarium speciespathogenic to cereals. European Journal of PlantPathology, 109: 755–768.

Publications



Dunne LA and Feehan J (2003). The origin and sig-nificance of mushroom stones in lowland karstregions. Irish Journal of Earth Sciences, 20: 33–40.

Feehan J (2003). Divine earth. A Christianperspective on nature. Resurgence, 221, 6–9.

Fournier N, Pais VA, Sutton MA, Weston KJ,Dragosits U, Tang SY and Aherne J (2002).Parallelisation and application of a multi-layeratmospheric transport model to quantifydispersion and deposition of ammonia over theBritish Isles. Environmental Pollution, 116: 95–107.

Gebauer J (2002). Effects of different nitrogencontents in wheat leaves on the food choice andfeeding activity of the grey field slug Derocerasreticulatum (Müller) under laboratoryconditions. Zeitschrift für Pflanzenkrankheitenund Pflanzenschutz – Journal of Plant Diseases andProtection, 109: 421–429.

Gebauer J (2002). Survival and food choice of thegrey field slug (Deroceras reticulatum) on threedifferent seed types under laboratoryconditions. Anzeiger für Schädlingskunde –Journal of Pest Science, 75: 1–5.

Gray JS (2002). Biology of Ixodes species ticks inrelation to tick-borne zoonoses. Wiener KlinischeWochenschrift, 114: 473–478.

Gray JS, von Stedingk LV and Granström M(2002). Zoonotic babesiosis. International Journalof Medical Microbiology, 291 (Suppl. 33): 108–111.

Gray JS, von Stedingk LV, Gürtelschmid M andGranström M (2002). Transmission studies onBabesia microti in Ixodes ricinus ticks and gerbils.Journal of Clinical Microbiology, 40: 1258–1263.

Hakvoort SGH and Schmidt O (2002). 15N stableisotope labelling of slugs (Gastropoda:Pulmonata). Annals of Applied Biology, 141:275–281.

Hassall M, Helden A and Benton T (2003).Phenotypic plasticity and interpopulationdifferences in life history of Armadillidium vulgare(Isopoda: Oniscidae). Oecologia, 137: 85–89.

Helden AJ and Dixon AFG (2002). Life-cyclevariation in the aphid Sitobion avenae: costs andbenefits of male production. EcologicalEntomology, 27: 692–701.

Hyde BP, Carton OT, O’Toole P and MisselbrookTH (2003). A new inventory of ammoniaemissions from Irish agriculture. AtmosphericEnvironment, 37: 55–62.

Langton C, Gray JS, Waters PF and Holman PJ(2003). Naturally-acquired babesiosis in areindeer (Rangifer tarandus tarandus) herd inGreat Britain. Parasitology Research, 89: 194–198.

McCabe OM and Morgan MA (2002). Phosphorusbehaviour on a grassland drumlin slope, 2:reseeded pasture. Tearmann, 2: 69–76.

McDonnell RM, Holden NM, Ward SM, Collins JF,Farrell EP and Hayes MHB (2001).Characteristics of humic substances inheathland and forested peat soils. Biology andEnvironment. In Press.

McNamara K, O’Kiely P, Whelan J, Forristal PD andLenehan JJ (2002). Preventing bird damage towrapped baled silage during short- and long-term storage. Wildlife Society Bulletin, 30: 809–815.

McNamara K, O’Kiely P, Whelan J, Forristal PDand Lenehan JJ (2002). Simulated bird damageto the plastic stretch-film surrounding baledsilage and its effects on conservation character-istics. Irish Journal of Agricultural and FoodResearch, 41: 29–41.

McNamara K, Whelan J, O’Kiely P, Forristal PDand Lenehan JJ (2002). Bird damage to theplastic stretch-film on baled silage in Ireland.Aspects of Applied Biology, 67: 112–120.

Muldowney J and Schmidt O (2002). Allolobophoracupulifera Tétry (Oligochaeta: Lumbricidae) inIreland: first records for the British Isles.Megadrilogica, 9: 29–32.

Muldowney J, Curry JP, O’Keeffe J and Schmidt O(2003). Relationships between earthwormpopulations, grassland management andbadger density in County Kilkenny, Ireland.Pedobiologia, 47: 913–919.

O’Donovan G (2002). Nutrient dynamics ofSesleria-dominated grasslands in the BurrenNational Park, Co. Clare. Tearmann, 2: 29–38.

O’Neill G and Whelan J (2002). The occurrence ofCorynosoma stromosum in grey seals caught ofthe Atlantic coast of Ireland. Journal ofHelminthology, 76: 231–234.

Pichon B, Egan D, Rogers M and Gray JS (2003).Detection and identification of pathogens andhost DNA in unfed host-seeking Ixodes ricinus L.(Acari: Ixodidae). Journal of Medical Entomology,40: 723–731.

Purvis G and Fadl AM (2002). The influence ofcropping rotations and soil cultivation practiceon the population ecology of carabids(Coleoptera: Carabidae) in arable land.Pedobiologia, 46: 452–474.

Purvis G, Chauzat MP, Segonds-Pichon A andDunne R (2002). Life history and phenology ofthe eucalyptus psyllid, Ctenarytaina eucalypti(Homoptera: Psylloidea) in Ireland. Annals ofApplied Biology, 141: 283–292.



Schmidt O, Clements RO and Donaldson G (2003).Why do cereal–legume intercrops support largeearthworm populations? Applied Soil Ecology,22: 181–190.

Schmidt O, Dyckmans J and Scrimgeour CM(2003). Isotopic labelling of earthworms:Comments on the paper by Whalen andJanzen (2002). Soil Biology and Biochemistry, 35:875–876.

Schrader S and Larink O (2003). Earthworms aspromoters of soil structure rehabilitation.Tearmann, 3: in press.

Spehn EM, Scherer-Lorenzen M, Schmid B,Hector A, Caldeira MC, Dimitrakopoulos PG,Finn JA, Jumpponen A, O’Donovan G, PereiraJS, Schulze ED, Troumbis AY and Korner C(2002). The role of legumes as a component ofbiodiversity in a cross-European study ofgrassland biomass nitrogen. Oikos, 98:205–218.

Torpey P, Morgan MA and McCabe OM (2002).Phosphorus behaviour on a grassland drumlinslope, 1: permanent pasture. Tearmann, 2: 53–67.

van Maanen A and Xu XM (2003). Modelling plantdisease epidemics. European Journal of PlantPathology, 109: 669–682.

Vor T, Dyckmans J, Loftfield N, Beese F and FlessaH (2003). Aeration effects on CO2, N2O, and CH4

emission and leachate composition of a forestsoil. Journal of Plant Nutrition and Soil Science,166: 39–45.

Xu XM, Parry DW, Nicholson P, Thomsett MA,Simpson D, Edwards SG, Cooke BM, Doohan F,van Maanen A, Moretti A, Tocco G, Mule G,Hornok L, Giczey G, Tatnell J and Ritieni A(2003). Is the amount of mycotoxins in cerealgrains related to the quantity of Fusarium DNA?Aspects of Applied Biology, 68: 101–108.

Zintl A, Westbrook C, Skerrett H, Gray JS andMulcahy G (2002). Chymotrypsin and neu-raminidase treatment inhibit host cell invasionby Babesia divergens (Phylum Apicomplexa).Parasitology, 125: 45–50.

Zintl A, Westbrook C, Skerrett H, Mulcahy G andGray JS (2002). Invasion and short- and long-term survival of Babesia divergens (PhylumApicomplexa) in non-bovine erythrocytes.Parasitology, 124: 583–588.

Zintl A, Mulcahy G, Skerrett HE, Taylor SM andGray JS (2003). Babesia divergens: a bovineblood parasite of veterinary and zoonoticimportance. Clinical Microbiology Reviews, 16:622–636.

Proceedings and AbstractsAnsari KI, Palacios N, Araya C, Langin T, Egan D

and Doohan FM (2002). Characterization ofColletotrichum lindemuthianum, the causalorganism of anthracnose disease of commonbean (Phaseolus vulgaris), using differentialcultivars and molecular marker analysis. 6thConference of the European Federation of PlantPathology, Prague, p. 67.

Ansari KI, Egan D, And Doohan FM (2003).Differential gene expression in four wheatcultivars in response to trichothecenemycotoxin deoxynivalenol. 11th InternationalCongress on Molecular Plant Microbe Interaction,St. Petersburg, Russia, p. 218.

Ansari KI, Rocha O, Egan D, McCabe P andDoohan FM (2003). Deoxynivalenol mycotoxin:Plant cell death and differential geneexpression. Irish Plant Scientists’ Meeting, Dublin,p. 26.

Bennett CM and Cooke BM (2002). Fungalpathogen development in conventional andweedy winter wheat. Proceedings of the VIIthCongress of the European Society for Agronomy,Cordoba, Spain, pp. 167–168.

Black K, Osborne B, Bolger T, Reidy B, TobinB, Gardiner J, Schmidt O, Dyckmans J,Byrne K, Saiz G and Farrell T (2003). Age-related dynamics of carbon exchange inIrish forests. Pathways to a SustainableFuture – A Showcase of EnvironmentalResearch. 10th Anniversary Conference of theEnvironmental Protection Agency, Dublin, p.58.

Brennan JM, van Maanen A, Egan D, Cooke BMand Doohan FM (2002). Effect of temperature ongrowth and in vitro pathogenicity of Europeanphytopathogenic Fusarium species causing headblight disease of wheat. 6th Fungal GeneticsConference, Pisa, Italy, p. 182.

Brennan JM, van Maanen A, Egan D, Cooke BMand Doohan FM (2002). PhytopathogenicFusarium fungi: effect of temperature on growthand in vitro pathogenicity of European Fusariumspecies causing head blight disease of wheat.Proceedings of the VIIth Congress of the EuropeanSociety for Agronomy, Cordoba, Spain, pp.171–172.

Brennan JM, van Maanen A, Egan D, Cooke BMand Doohan FM (2003). Effect of temperature ongrowth and in vitro pathogenicity of Europeanphytopathogenic Fusarium species. 8thInternational Congress of Plant Pathology,Christchurch, New Zealand, p. 208.



Brennan JM, van Maanen A, Leonard G, Fagan B,Cooke BM and Doohan FM (2003). Fusariumhead blight: a comparison of in vitro and in vivotechniques for screening pathogenicity anddisease resistance. IXth International FusariumWorkshop, Sydney, Australia, p. 24.

Brennan J, van Maanen A, Leonard G, Fagan B,Cooke BM and Doohan FM (2003). Screeningfungal pathogenicity and host disease resistancefor Fusarium head blight disease in Europe. IrishPlant Scientists’ Meeting, Dublin, p. 27.

Byrne KA, Black K, Bolger T, Farrell EP, GardinerJJ, McCullagh S, Osborne BO, Saiz G, Reidy B,Schmidt O and Tobin B (2002). A study tocompare inventory and flux based methodolo-gies for measuring carbon sequestration inforest ecosystems. 2nd CarboEurope Meeting,Budapest, Hungary.

Coulahan P and Whelan J (2003). Operationalinteractions between seals and salmon (Salmosalar) summer drift net fisheries on the WestCoast of Ireland. 4th European Vertebrate PestManagement Conference, Parma, Italy, p. 37.

Curry JP and Schmidt O (2002). Plenary Talk:Linking soil invertebrate biodiversity with soilprocesses. Workshop Kommission III Bodenbiologieder Deutschen Bodenkundlichen Gesellschaft,Neuherberg/Munich, Germany. Mitteilungen derDeutschen Bodenkundlichen Gesellschaft, 99: 167.

Doohan FM, Brennan J, van Maanen A, McCann J,Egan D and Rocha O (2002). Effect of temperatureon Fusarium fungi: growth and heat shock geneexpression. 6th Conference of the EuropeanFederation of Plant Pathology, Prague, p. 79.

Doohan FM, Brennan J, van Maanen A, McCann J,Egan D and Rocha O (2002). Effect oftemperature on Fusarium fungi: growth andheat shock gene expression. 6th Fungal GeneticsConference, Pisa, Italy, p. 436.

Doohan FM, Brennan J, van Maanen A, Rocha O,Ansari K, Tuohy J, Cooke, BM and Egan D(2003). Fusarium head blight of wheat in Ireland:studies on incidence, pathogenicity and hostsusceptibility. IXth International FusariumWorkshop, Sydney, Australia, p. 21.

Doohan FM, Rocha O, Ansari K and Egan D(2003). Understanding the host-mycotoxininteraction. Irish Plant Scientists’ Meeting,Dublin, p. 15.

Hyde BP, Misselbrook T, Carton OT and O’Toole P(2003). The Irish ammonia emission inventory :implications for compliance with theGothenburg Protocol. 12th International NitrogenCycling Workshop, Exeter, UK.

Khan MR, Egan D and Doohan FM (2003).Culture-dependent microbial diversity in thecereal growing soils of Ireland – a preliminarystudy. Irish Plant Scientists’ Meeting, Dublin, p.36.

McCabe, OM, McDonald, PC and Morgan, MA(2002). Occurrence and phosphorus concentra-tions of surface runoff from a drumlin soil.Proceedings 25th Annual Research Meeting of theSoil Science Society of Ireland, Tullamore, p. 8.

McCabe OM, McNeilis N and Morgan, MA (2002).Differences in adsorption and desorption ofphosphorus among mineral soils. 12th IrishEnvironmental Researchers’ Colloquium, Cork, p.58.

McNamara K, O’Kiely P, Whelan J, Forristal PDand Lenehan JJ (2002). Bird damage to theplastic stretch-film on baled silage: theculprits. Proceedings of the XIIth InternationalSilage Conference, Auchincruive, UK, pp.166–167.

McNamara K, O’Kiely P, Whelan J, Forristal PDand Lenehan JJ (2002). Control of bird damageto the plastic stretch-film surrounding baledsilage. Proceedings of the XIIth InternationalSilage Conference, Auchincruive, UK, pp.150–151.

Muldowney J, Curry JP and Schmidt O (2002).Relationships between earthworm populations,grassland management and badger densities inCo. Kilkenny, Ireland. 7th InternationalSymposium on Earthworm Ecology, Cardiff, UK, p.377.

Murphy J, Egan D, Leonard G and Doohan FM(2003). Fusarium head blight of wheat andbarley. Irish Plant Scientists’ Meeting, Dublin, p.34.

O’Meara CO, Egan D, Doohan FM and EvansACO (2002). Differential gene expression indominant and subordinate bovine ovarianfollicles using DDRT-PCR. 1st Conway InstituteFestival of Research, Dublin, p. 90.

Quilter JM, Schmidt O, Monahan FJ, Boland MP,Cooke DW and Scrimgeour CM (2002).Carbon and nitrogen stable isotope ratios incattle: variation within and between musclesand populations. 3rd International Conferenceon Applications of Stable Isotope Techniques toEcological Studies, Flagstaff, Arizona, USA, p.130.

Rocha O, Egan D, Heffernan A and Doohan FM(2002). Cytological and molecular response ofwheat tissue to mycotoxin treatment. IrishBotanists’ Meeting, Maynooth, p. 41.



Ruckenbauer P, Borum F, Neumayer A, Weyen J,Mesterhazy A, Doohan FM, Cooke BM, Adam Gand Nicholson P (2003). Novel tools fordeveloping Fusarium-resistant toxin-free wheatfor Europe: the EU FUCOMYR project. 8thInternational Congress of Plant Pathology,Christchurch, New Zealand, p. 291.

Schmidt O (2002). Earthworm populations underconventional and reduced tillage: a literaturemeta-study. 7th International Symposium onEarthworm Ecology, Cardiff, UK, p. 330.

Schmidt O, Curry JP and Scrimgeour CM (2002).Nitrogen and carbon assimilation from surfaceplant litter by undisturbed earthworm(Lumbricidae) populations. 7th InternationalSymposium on Earthworm Ecology, Cardiff, UK, p.285.

Schmidt O, Dyckmans J, Scrimgeour CM and CurryJP (2002). Intestinal secretion, reabsorption andloss of carbon and nitrogen in lumbricidearthworms. 7th International Symposium onEarthworm Ecology, Cardiff, UK, p. 331.

Schmidt O, Monahan FJ, Dunne PG, Moloney AP,Scrimgeour CM and Begley IS (2002). Inferringthe dietary history of beef from light elementstable isotope ratios. 3rd International Conferenceon Applications of Stable Isotope Techniques toEcological Studies, Flagstaff, Arizona, USA, p. 143.

Schmidt O, Curry JP, Rota E and Scrimgeour CM(2003). Dual stable isotope analysis (δ13C andδ15N) of a detritivore soil food web. BritishEcological Society 2003 Annual Symposium,Lancaster, UK, p. 22.

Schmidt O, Dyckmans J and Black K (2003).Bodenalgen als C-Quelle für Bodentiere. Tagungder Deutschen Bodenkundlichen Gesellschaft,Franfurt/Oder, Germany. Mitteilungen derDeutschen Bodenkundlichen Gesellschaft, 102:397–398.

Sheridan H, Culleton N and O’Donovan G (2003).Field margin management to enhance biodiver-sity on grazing land. 13th Irish EnvironmentalResearchers’ Colloquium, Galway, p. 57.

Sheridan H, Culleton N and O’Donovan G (2003).The Syrphidae (Diptera) of contrastinggrassland farms in Ireland. II InternationalSymposium on the Syrphidae – Biodiversity andConservation, Alicante, Spain, p. 33.

Sticht S, Schrader S, Giesemann A, Larink O andWeigel HJ (2003) Effects of atmospheric CO2

enrichment on collembolan biodiversity and C-isotopic composition in an agroecosystem.British Ecological Society 2003 Annual Symposium,Lancaster, UK, p. 31.

Tuohy J, Jalli M, Cooke BM and O’Sullivan E(2002). Pathogenic variation in populations ofDrechslera teres f.sp. teres and Drechslera teres f.sp. maculata. Proceedings of the VIIth Congress ofthe European Society for Agronomy, Cordoba,Spain, pp. 229–230.

Tuohy J, Egan D, Murray C and Doohan FM (2003).Isolation and expression analysis of a b-glucosidasegene from the wilt pathogen Fusarium oxysporum.Irish Plant Scientists’ Meeting, Dublin, p. 36.

van Maanen A, Leonard G, Brennan JM, DoohanFM and Cooke BM (2003). Disease epidemiolo-gy of Fusarium head blight in Ireland: influenceof climatic factors on the incidence of FHB inwheat during 2001 and 2002. Irish PlantScientists’ Meeting, Dublin, p. 36.

van Maanen A, Leonard G, Brennan JM, DoohanFM and Cooke BM (2003). Influence of climaticfactors on the incidence of Fusarium Head Blight(FHB) and mycotoxin in wheat across Irelandduring 2001 and 2002. Abstracts of the 8thInternational Congress of Plant Pathology,Christchurch, New Zealand, p. 106.

ReportsHealth Research Board (2003). Health and environ-

mental effects of landfilling and incineration of waste– A literature review. Health Research Board,Dublin (Dr. John Fry contributions to, andediting of, section on Environmental Effects).

Scott S, Bacon P and Fry J (2003). Evaluation of Eco-Auditing in the context of the National DevelopmentPlan 2000–2006. Report prepared for theNDP/CSF Evaluation Unit, Department ofFinance, Dublin.

BooksDunne L and Feehan J (2003). Ireland’s Mushroom

Stones: Relics of a Vanished Lakeland. UCD,Department of Environmental ResourceManagement.

Feehan J (2003). Farming in Ireland: History, Heritageand Environment. UCD Faculty of Agriculture,606 pp.

Edited BooksBailey JA, Logrieco A and Cooke BM (eds.) (2002).

Mycotoxins in Plant Disease. Kluwer AcademicPublishers, Dordrecht, the Netherlands. 176 pp.

Convery F and Feehan J (2003). Achievement andChallenge: Rio+10 and Ireland. UCD, TheEnvironmental Institute, 580 pp.



Gray JS, Kahl O, Lane RS and Stanek G (eds.)(2002). Lyme Borreliosis: Biology, Epidemiology andControl. CAB International, Wallingford, UK.347 pp.

Xu XM, Bailey JA, and Cooke BM (eds.) (2003).Epidemiology of Mycotoxin Producing Fungi.Kluwer Academic Publishers, Dordrecht, theNetherlands. 136 pp.

Book ChaptersBolger T, Schmidt O, Purvis G and Curry JP (2002).

The biodiversity, function and management ofsoil invertebrate populations – an Irishperspective. In “Achievement and Challenge:Rio+10 and Ireland” edited by F Convery and JFeehan. The Environmental Institute,University College, Dublin, pp. 2–10.

Byrne KA and Farrell EP (2002). Towards andunderstanding of the role of Irish forests inmitigating greenhouse gas emissions. In“Achievement and Challenge: Rio+10 and Ireland”edited by F Convery and J Feehan. TheEnvironmental Institute, University College,Dublin, pp. 112–115.

Convery F, Fry J, Matthews A, O’Shea S andPender A (2002). European Union agri-environ-mental policy: Issues and potential. In “Greeningthe budget: Budgetary policies for environmentalimprovement” edited by JP Clinch, KSchlegelmilch, RU Sprenger and UTriebswetter. Edward Elgar, Cheltenham, UK,pp. 248–274.

Farrell EP (2001). Ecological impacts of plantationforests on water and nutrient cycles andecosystem stability. In “Ecological and Socio-Economic Impacts of Close-to-Nature Forestry”edited by T Green. EFI Proceedings, 37, pp.27–34.

Farrell EP and Byrne KA (2002). The emergence ofthe multifunctional forest in Ireland. In“Achievement and Challenge: Rio+10 and Ireland”edited by F Convery and J Feehan. TheEnvironmental Institute, University College,Dublin, pp. 18–22.

Feehan J (2002). Biodiversity and Ireland: meetingthe challenge of the Convention. In“Achievement and Challenge: Rio+10 and Ireland”edited by F Convery and J Feehan. TheEnvironmental Institute, University College,Dublin, pp. 23–28.

Feehan J (2002). Towards multifunctionalpeatlands. In “Peat in Horticulture. Quality andEnvironmental Challenges” edited by G.Schmilewski and L. Rochefort. Proceedings ofthe International Peat Symposium, PärnuEstonia. International Peat Society, pp. 171–177.

Feehan J (2002). Heritage inside peat: exploringthe heritage value of Ireland’s boglands. In“Local and global heritage, Proceedings of theHeritage Seminars at Koli National Park andNational Landscape in Finland, August 2001,”edited by Lasse Loven, pp. 51–64.

Feehan J (2001). Using the rural environment as aresource for rural development. Tessedik SámuelFöiskola Tudományos közlemenyek (ScientificJournal of Tessedik Samuel College) 1 (2), pp.305-310.

Feehan J (2002). Urban Nature and HumanNature. In “Biodiversity in the City” edited by LDunne. UCD, Environmental Institute, pp. 6–9.

Feehan J (2003). Creation as Revelation: a newethic towards the living world. In “A JustSociety? Ethics and Values in ContemporaryIreland” edited by J. Scally. Dublin, The LiffeyPress, pp. 93–102.

Hyde BP, Carton OT and O’Toole P (2002).Ammonia emissions from Irish agriculture. In“Achievement and Challenge: Rio+10 and Ireland”edited by F Convery and J Feehan. TheEnvironmental Institute, University College,Dublin, pp. 52–55.

Stanek S, Strle F, Gray J and Wormser G (2002).History and characteristics of Lyme borreliosis.In “Lyme Borreliosis: Biology, Epidemiology andControl” edited by JS Gray, O Kahl, RS Lane andG Stanek. CAB International, Wallingford, UK,pp. 1–28.

Whelan J and Butler F (2002). Introduced rodents:their significance in Ireland. In “BiologicalInvaders: The Impact of Exotic Species” edited by CMoriarty and D Murray. Royal Irish Academy,Dublin, pp. 84–89.

Other (electronic media)Gray JS (2003). Borrelia burgdorferi in livestock. In

“Animal Health Compendium”. CABInternational, Wallingford, UK (available asCD–ROM and online at http://www.ani-malscience.com).



Bennett CM (2002). Stem-base and foliar diseasedevelopment in diversified wheat stands.

Hackett RA (2003) Aspects of productivity inwheat growing in a clover understorey.

McNamara KM (2002). Prevention of vertebratepest damage to the plastic stretch-filmsurrounding baled silage.

Qu X (2001). Molecular characterisation anddetection of Spongospora subterranea F. sp.subterranea, the cause of powdery scab of potato.

Shafizadeh S (2003) Phytophthora root rot-stemcanker of Noble Fir christmas trees in Ireland.

Tuohy J (2002). Comparison of the net- and spot-forms of Drechslera teres on barley.

Doctoral Theses

Molloy AM (2002). Molecular and physiologicalresponses of pedunculate oak (Quercus robur)seedlings to heat stress.

Quilter JM (2002). Determination of the dietaryhistory and geographical origin of bovinemuscle using stable isotope analysis.

Roarty SP (2003). Developing an EnvironmentalHeritage Evaluation and Panagement Plan forLyons Research Farm.

Westwood C (2002). In vitro culturing andattenuation of a potential live vaccine againstbovine babesiosis.

Masters Theses

Other Completed ProjectsHost resistance to Fusarium ear blight of wheat andthe associated mycotoxin contamination of grainFM Doohan, O Rocha and D EganFunded by Enterprise Ireland Basic Research Grant

Study of the genetic variability of the pathosys-tem common bean anthracnose and identifica-tion of durable resistance sources to reducebean yields in Latin America and AfricaFM Doohan, BM Cooke, J Tuohy and D EganFunded by the EU INCO-DC Programme

Agriculturally important toxigenic fungiFM Doohan and BM CookeCOST Action 835 funded by the EuropeanCommission

Common molecular events during tissue deathin plant and animal cellsACO Evans*, CJ O’Reilly**, FM Doohan, CO`Mara, AM Molloy, K Ansari and D Egan*Department of Animal Science and Production** Department of Crop Science, Horticulture andForestryFunded by Faculty of Agriculture Research Award

Determination of the origin and dietary back-ground of beef using stable isotope analysisFM Monahan*, O Schmidt and J Quilter* Department of Food ScienceFunded by Faculty of AgricultureInterdepartmental Studentship

Projects in ProgressMonitoring, functional significance and manage-ment for the maintenance and economic utiliza-tion of bio-diversity in the intensively farmedlandscape (Ag-Biota)G Purvis, JP Curry, O Schmidt, J Whelan, AAnderson, A Helden, B McMahon, D Miksche, TBolger*, M Kelly-Quinn*, J Connolly**, J Breen***,J Finn****, R Schulte****, T Kennedy**** and others*Department of Zoology, UCD, ** Department of

Statistics, ***University of Limerick, ****TeagascFunded by the Environmental Protection Agency

The role of field margins in the farmed land-scape – form, function and biodiversity aspectsG O’Donovan, N Culleton* and H Sheridan*TeagascFunded by a Teagasc Walsh Fellowship



Sustainability, product safety and quality incereals: development of novel quantitative mod-els for risk assessment of mycotoxigenicFusarium species (RAMFIC)BM Cooke, FM Doohan, A van Maanen, JMBrennan, D Egan and othersFunded under the EU 5th Framework Programme

Novel tools for developing Fusarium resistantand toxin-free wheat for Europe (FUCOMYR)FM Doohan, BM Cooke, K Ansari, R Browne andD EganFunded under the EU 5th Framework Programme

Cereal pathology: improving the genetic basis ofhost resistance to fungal diseasesFM Doohan, J Murphy, D Egan and othersFunded by a DAFF Research Stimulus Grant

Reduction of fungicide input in Irish cerealfarming: biological control of cereal foliar andhead blight fungal pathogensFM Doohan, M Khan, S Kildea, D Egan and othersFunded by a DAFF Research Stimulus Grant

Development of a risk assessment assay for the pow-dery scab/spraing complex of potatoes using PCRFM Doohan, J Tuohy, J Kavanagh and D EganFunded by a DAFF Research Stimulus Grant

Study of nutrient value of Irish biosolidsW Kato*, OT Carton* and P O’Toole* TeagascFunded by Department of the Environment andLocal Government

Methane and nitrous oxide emission factors:emission factors for nitrous oxideO Carton*, M Ryan*, B Hyde*, A Fanning*, FO’Mara** and P O’Toole*Teagasc, Johnstown Castle, ** Department ofAnimal Science and Production, UCDFunded by the Environmental Protection Agency

A study on the biodiversity of farmland birds inrelation to habitatB McMahon and J WhelanFunded by the Environmental Protection Agency

Behaviour and range of the Hen Harrier (Circuscyaneus)B O’Donoghue and J WhelanFunded by Faculty of Agriculture, and Parks andWildlife Service

The value of parasitic Hymenoptera as indicatorsof biological diversity in agro-ecosystems G Purvis and A AndersonFunded by Environmental Protection Agency

Use of entomopathogenic fungi and a novelchemical delivery system to control soil pests ofhorticultural crops G Purvis, R Dunne*, M Maher* and M Gaffney*TeagascFunded by a Teagasc Walsh Fellowship

PEnrich - Forestry Operations and EutrophicationOM McCabe, EP Farrell and P O’Dea**CoillteFunded by the EPA and COFORD

Determination of the origin and dietary back-ground of beefFJ Monahan*, O Schmidt, AP Moloney**, and BBahar****Department of Food Science, UCD, **Teagasc,***ERM and Department of Food Science, UCDFunded by a Teagasc Walsh Fellowship

Investigations into the use of winter covercrops in spring barley production systems inIrelandO Schmidt, RA Hackett* and E O’Keeffe*TeagascFunded by a Teagasc Walsh Fellowship

Forest entomology inventory for sustainable for-est managementO Schmidt, C O’Reilly* and R Gleeson*Department of Crop Science, Horticulture andForestry, UCDFunded by Faculty of Agriculture InterdepartmentalStudentship

Carbon sequestration in Irish forest ecosystems(CARBiFOR)EP Farrell, O Schmidt, M Nieuwenhuis*, TBolger**, B Osborne***, G Saiz and others*Department of Crop Science, Horticulture andForestry, UCD, **Department of Zoology, UCD,***Department of Botany, UCDFunded by COFORD

Documents

Environmental Resource Management · Deoxynivalenol (DON) is a toxic compound produced by cereal-pathogenic Fusarium fungi. DON is harmful to both animal and plant tissues. Evidence