DOI: 10.5281/zenodo.7307470
title: Soil biological, chemical and physical parameters and herbage yield in a field experiment with organic and inorganic fertilizers on peat grassland in the Netherlands

description: To evaluate the performance of organic and inorganic fertilizers for regeneration of ecosystem services in peat grasslands with biodiversity goals, we carried out a field experiment in the western peat district in the Netherlands. The fertilizers tested represent the current practice and potential alternatives for regenerative grassland management on drained peat. <strong>Experimental setup</strong> The field experiment (2013 – 2015) was conducted on a permanent grassland on peat soil (Terric Histosol; SOM 56 g 100 g<sup>−1</sup> and pH<sub>KCl</sub> of 4.5 in 0-10 cm) at the experimental dairy farm at Zegveld (the Netherlands). In March 2013, a randomized block experiment (six blocks) was laid out with six fertilizer treatments and a control treatment (no fertilizer: “Contr”). The fertilizer used were: conventional dairy cattle slurry manure (“Slurry”), mature compost of kitchen and garden waste (“Comp”), dairy cattle farmyard manure (“FYM”), solid fraction of the cattle slurry manure (“SFrac”, obtained by pressurized filtration), inorganic N fertilizer (“IF”; calcium ammonium nitrate, 27% N) and a combination of inorganic N fertilizer and sawdust (“IF+SD”). Plot size was 4 × 10 m; for the Slurry treatment plots were 5.2 × 10 m. Slurry was applied by slit injection, the other fertilizers were applied by hand. Target application rate was 120 kg total N ha<sup>−1</sup> yr<sup>−1</sup>, divided in two applications per year (February/March and May). This is relatively low for conventional grasslands but usual for grasslands with biodiversity goals (Kleijn et al., 2004). The amount of C<sub>total</sub> applied in Comp was taken for the rate of sawdust to be applied. All plots were fertilized with 200 kg K<sub>2</sub>O ha<sup>−1</sup> yr<sup>−1</sup> (applications in March and May) (Commissie Bemesting Grasland en Voedergewassen, 2019). Fertilizer application quantities and organic matter and nutrient inputs are provided in Fertilizer_intput.csv (dataset). The grassland had an history of conventional management with mainly cutting, winter grazing with sheep and a normal fertilization regime with both slurry manure and inorganic fertilizer. The normal cutting and grazing regime was continued in the first two years of the experiment; during 2015, the monitoring year, the plots were not grazed and only cut for herbage measurements. <strong>Measurements</strong> From April to October 2015, soil and aboveground measurements were carried out. Most soil parameters were measured in October. Earthworms and insect larvae are an important food source for meadow birds during the pre-breeding period in spring (Galbraith, 1989) and were therefore sampled in April. Soil moisture and penetration resistance were measured both in April and October. <em>Soil biological parameters</em> Earthworms and insect larvae were sampled in the top soil layer in two soil cubes (20 × 20 × 20 cm) per plot. Earthworms were hand-sorted, counted, weighed and fixed in alcohol prior to identification. Both adults and juveniles were identified to species (Sims and Gerard, 1985; Stöp-Bowitz, 1969) and classified into functional groups (Bouché, 1977). Crane flies (Tipulidae; leatherjackets) or click beetles (Elateridae; wireworms) larvae were counted. Phospholipid fatty acids (PLFA) were measured in October. PLFA were extracted from 4 g of fresh soil (Palojärvi, 2006), and analyzed by gas chromatography (Hewlett-Packard, USA). PLFA i15:0, a15:0, 15:0, i16:0, 16:1ω9, i17:0, a17:0, cy17:0, 18:1ω7 and cy19:0 were chosen to represent bacteria and PLFA 18:2ω6 was used as a marker of saprotrophic fungi (Hedlund, 2002). The neutral lipid fatty acid (NLFA) 16:1ω5 occurs in storage lipids of arbuscular mycorrhizal fungi (AMF) and was used as marker of AMF (Vestberg et al., 2012). PLFA i15:0, a15:0, i16:0, i17:0 and a17:0 were used as a measure of Gram-positive bacteria, and cy17:0 and cy19:0 for Gram-negative bacteria. PLFA 10Me16:0, 10Me17:0 and 10Me18:0 represented actinomycetes. <em>Soil chemical parameters</em> A soil sample from the 0−10 cm layer (c. 50 randomly taken soil cores) per experimental plot was collected in October (auger diameter 2.3 cm; Eijkelkamp grass plot sampler, Giesbeek, the Netherlands), was sieved (1 cm mesh size) and homogenized. One sub-sample was taken for analysis of hot water extractable carbon (HWC) according to Ghani et al. (2003) and one for chemical analysis. Prior to analysis of soil acidity (pH<sub>KCl</sub>), soil organic matter (SOM), total carbon (C<sub>total</sub>), total nitrogen (N<sub>total</sub>), total phosphorus (P<sub>total</sub>) and ammonium-lactate extractable P (P<sub>AL</sub>) by Eurofins Agro (Wageningen, the Netherlands), the sub sample was dried at 40°C. Soil pH<sub>KCl</sub> was measured according to NEN-ISO 10390 2005. SOM was determined by loss-on-ignition (NEN 5754 2005). C<sub>total</sub> was measured by incineration at 1150°C, and determination of the CO<sub>2</sub> produced by an infrared detector (LECO Corporation, St. Joseph, Mich., USA). For N<sub>total</sub>, evolved gasses after incineration were reduced to N<sub>2</sub> and measured with a thermal-conductivity detector (LECO Corporation, St. Joseph, Mich., USA). P<sub>total</sub> was analysed with Fleishmann acid (Houba et al., 1997). P<sub>AL</sub> is used to assess the P supply capacity of grassland soils (Reijneveld et al., 2014) and was determined according to Egnér et al. (1960) (NEN 5793). <em>Soil physical parameters</em> Soil moisture was determined in April and October in a homogenized 0−10 cm soil sample after drying at 105°C for 24 hrs. Moisture content was expressed as percentage of fresh soil weight. Penetration resistance was measured (April and October) with a penetrologger (Eijkelkamp, Giesbeek, the Netherlands; cone of 2.0 cm<sup>2</sup> penetration surface and 60° apex angle. Penetration resistance was expressed as an average of 7 penetrations per plot and per soil layer of 0−10, 10−20, and 20−30 cm. Soil structure and rooting density were assessed in October in the 0−10 cm and 10−25 cm layers. The percentage of crumbs, sub-angular blocky elements and angular blocky elements was estimated by one experienced person as described by Peerlkamp (1959) and Shepherd (2000), Root density was estimated by scoring visible roots (score 1–10; 1 for no roots and 10 for above average). Water infiltration rate was measured in October at three spots per experimental plot in 5 of the 6 blocks (35 plots). A PVC pipe (15 cm high, 15 cm diameter) was pushed into the soil to a depth of 10 cm. 500 ml water was poured into each pipe and the infiltration time was recorded. If the infiltration time exceeded 15 min, the remaining water volume was estimated to calculate the infiltration rate (mm min<sup>−1</sup>). <em>Grass yield and botanical composition</em> Grass dry matter (DM) and N yield were determined during 2015 with a Haldrup plot harvester (J. Haldrup a/s, Løgstør, Denmark). The four harvest dates were May 15, June 29, August 19 and September 30. Fresh biomass, DM content (70°C for 24 hrs) and total N content (Kjeldahl) were determined for each harvest. Herbage DM yield (Mg DM ha<sup>−1</sup>) and herbage N yield (kg N ha<sup>−1</sup>) were calculated. Apparent N recovery (ANR; kg N.kg N<sup>−1</sup>) was calculated as (N yield<sub>(fertilized)</sub> – N yield<sub>(non-fertilized)</sub>)/(N fertilization rate) (Vellinga and André, 1999). In June 2015, botanical composition was measured by visually estimating the relative soil cover of the sward and the proportion of each species therein (Sikkema, 1997). <strong>Data files</strong> <em><strong>Data_soil_grass.csv</strong></em> <em>Content:</em> Dataset with soil biological (earthworms, microbial PLFA), soil chemical, soil physical parameters, herbage dry matter and N yields, and botanical parameters. <em>Column names and units:</em> plot: Experimental plot number (1-42) treatment: Treatment code (see text) block: Block number (1-6) EW_species_number: Earthworm - number of species EW_totalnumber: Earthworm - total number per m2 EW_epigeic: Earthworm - number of epigeic adults and juveniles per m2 EW_endogeic: Earthworm - number of endogeic adults and juveniles per m2 EW_adults: Earthworm - number of adults per m2 EW_juveniles: Earthworm - number of juveniles per m2 EW_adult_epigeic: Earthworm - number of epigeic adults per m2 EW_adult_endogeic: Earthworm - number of endogeic adults per m2 EW_juven_epigeic: Earthworm - number of epigeic juveniles per m2 EW_juven_endogeic: Earthworm - number of endogeic juveniles per m2 EW_L_rubellus: Earthworm - number of L. rubellus adults and juveniles per m2 EW_A_chlorotica: Earthworm - number of A. chlorotica adults and juveniles per m2 EW_A_caliginosa: Earthworm - number of A. caliginosa adults and juveniles per m2 EW_O_lacteum: Earthworm - number of O. lacteum adults and juveniles per m2 EW_A_rosea: Earthworm - number of A. rosea adults and juveniles per m2 EW_O_cyaenum: Earthworm - number of O. cyaneum adults and juveniles per m2 EW_L_castaneus: Earthworm - number of L. castaneus adults and juveniles per m2 EW_D_rubida: Earthworm - number of D. rubida adults and juveniles per m2 EW_adult_L_rubellus: Earthworm - number of L. rubellus adults per m2 EW_adult_A_chlorotica: Earthworm - number of A. chlorotica adults per m2 EW_adult_A_caliginosa: Earthworm - number of A. caliginosa adults per m2 EW_adult_O_lacteum: Earthworm - number of O. lacteum adults per m2 EW_adult_A_rosea: Earthworm - number of A. rosea adults per m2 EW_adult_O_cyaenum: Earthworm - number of O. cyaneum adults per m2 EW_adult_L_castaneus: Earthworm - number of L. castaneus adults per m2 EW_adult_D_rubida: Earthworm - number of D. rubida adults per m2 EW_juven_L_rubellus: Earthworm - number of L. rubellus juveniles per m2 EW_juven_A_chlorotica: Earthworm - number of A. chlorotica juveniles per m2 EW_juven_A_caliginosa: Earthworm - number of A. caliginosa juveniles per m2 EW_non_determined: Earthworm - number of non determined individuals per m2 EW_total_biomass: Earthworm - total fresh biomass per m2 Leatherjackets: number of leatherjackets per m2 Wireworms: number of wireworms per m2 TOTmicrPLFA: total microbial PLFA in nmol.g-1 dry soil bactPLFA: bacterial PLFA in nmol.g-1 dry soil saprofungPLFA: saprotrophic fungal PLFA in nmol.g-1 dry soil Fung_bactPLAF_ratio: ratio of fungal to bacterial PLFA GramPLUSplfa: gram positive PLFA in nmol.g-1 dry soil GramMINplfa: gram negative PLFA in nmol.g-1 dry soil ratioGram_PLUS_MIN: ratio of gram positive to gram negative PLFA AMFsporNLFA: AMF spores NLFA in nmol.g-1 dry soil ActinomPLFA: Actinomycetes PLFA in nmol.g-1 dry soil ShannonPLFA: PLFA shannon diversity index SOM: soil organic matter in g.100 g-1 dry soil Ctotal: total C in g.100 g-1 dry soil HWC: hot water extractable C in μg.100 g-1 dry soil Ntotal: total N in g.100 g-1 dry soil Ptotal: total P2O5 in mg.100 g-1 dry soil P_AL: total P-AL in mg.100 g-1 dry soil pH_KCl: pH-KCl CN_ratio: C:N ratio C_SOM: C:SOM ratio Soilmoisture_April: soil moisture content in April in g.100g-1 fresh soil Penetrationresistance_April_cm010: penetration resistance in April in 10-20 cm in Newton Penetrationresistance_April_cm1020: penetration resistance in April in 20-30 cm in Newton Penetrationresistance_April_cm2030: penetration resistance in April in 0-10 cm in Newton Soilmoisture_October: soil moisture content in October in g.100g-1 fresh soil Penetrationresistance_October_cm010: penetration resistance in October in 10-20 cm in Newton Penetrationresistance_October_cm1020: penetration resistance in October in 20-30 cm in Newton Penetrationresistance_October_cm2030: penetration resistance in October in 0-10 cm in Newton crumb_struct_cm010: percentage of crumb elements in 0-10 cm round_struct_cm011: percentage of sub-angular elements in 0-10 cm rootdensity_cm010: score (1-10) of root density in 0-10 cm crumb_struct_cm1025: percentage of crumb elements in 10-25 cm round_struct_cm1025: percentage of sub-angular elements in 10-25 cm sharp_struct_cm1025: percentage of angular elements in 10-25 cm rootdensity_cm1025: score (1-10) of root density in 10-25 cm water_infiltration: water infiltration rate in mm per minute DM_yield_year: total herbage dry matter yield in kg.ha-1 per year DM_yield_H1: herbage dry matter yield of harvest 1 in kg.ha-1 DM_yield_H2: herbage dry matter yield of harvest 2 in kg.ha-1 DM_yield_H3: herbage dry matter yield of harvest 3 in kg.ha-1 DM_yield_H4: herbage dry matter yield of harvest 4 in kg.ha-1 N_yield_year: total herbage N yield in kg.ha-1 per year N_yield_H1: herbage N yield of harvest 1 in kg.ha-1 N_yield_H2: herbage N yield of harvest 2 in kg.ha-1 N_yield_H3: herbage N yield of harvest 3 in kg.ha-1 N_yield_H4: herbage N yield of harvest 4 in kg.ha-1 DMperc_yield_year: herbage dry matter content (per year; weighed average over the 4 harvests) in g.100g-1 fresh weight DMperc_yield_H1: herbage dry matter content of harvest 1 in g.100g-1 fresh weight DMperc_yield_H2: herbage dry matter content of harvest 2 in g.100g-1 fresh weight DMperc_yield_H3: herbage dry matter content of harvest 3 in g.100g-1 fresh weight DMperc_yield_H4: herbage dry matter content of harvest 4 in g.100g-1 fresh weight Ncontent_yield_year: herbage N content (per year; weighed average over the 4 harvests) in g.kg-1 dry matter Ncontent_yield_H1: herbage N content of harvest 1 in g.kg-1 dry matter Ncontent_yield_H2: herbage N content of harvest 2 in g.kg-1 dry matter Ncontent_yield_H3: herbage N content of harvest 3 in g.kg-1 dry matter Ncontent_yield_H4: herbage N content of harvest 4 in g.kg-1 dry matter fresh_yield_H1: herbvage fresh yield of harvest 1 in Mg.ha-1 ANR: apparent N recovery in kg N.kg N-1 productive_grasses: cover percentage of L. perenne and P trivialis monocotyledons: cover percentage of monocotyledons dicotyledons: cover percentage of dicotyledons plant_species: number of plant species monocot_species: number of monocotyledon species dicot_species: number of dicotyledon species Lolium_perenne: plant cover % Poa_trivialis: plant cover % Phleum_pratense: plant cover % Elytrigia_repens: plant cover % Poa_annua: plant cover % Agrostis_stolonifera: plant cover % Holcus_lanatus: plant cover % Alopecurus_pratensis: plant cover % Alopecurus_geniculatus: plant cover % Trifolium_repens: plant cover % Taraxacum_officinale: plant cover % Ranunculus_arvensis: plant cover % Rumex_obtusifolius: plant cover % Rumex_crispus: plant cover % Ranunculus_acris: plant cover % Stellaria_media: plant cover % Cardamine_pratensis: plant cover % Bellis_perennis: plant cover % Rumex_acetosa: plant cover % Ranunculus_sceleratus: plant cover % Polygonum_aviculare: plant cover % Capsella_bursa-pastoris: plant cover % Glechoma_hederacea: plant cover % Geranium_molle: plant cover % <em><strong>Fertilizer_input.csv</strong></em> <em>Content:</em> Application quantities of fertilizers and ash, organic matter, C and mineral inputs, and fertilizer C:N ratio. Total N input is the sum of mineral N (Nmin) and organic N (Norg). Average values per hectare and per year over the years 2013−2015. <em>Column names and units:</em> Treatment: Treatment code (see text) Fertilizer_fresh: Applied fertilizer in Mg.ha<sup>-1</sup> per year (fresh weight) Fertilizer_DM: Applied fertilizer in Mg.ha<sup>-1</sup> per year (dry matter weight); for IF+SD this is the sum of 2.72 Mg sawdust + 0.45 Mg N fertilizer Ash: Mineral fraction in kg.ha<sup>-1</sup> per year OM: Organic matter in kg.ha<sup>-1</sup> per year C: Total C in kg.ha<sup>-1</sup> per year Nmin: Mineral N in kg.ha<sup>-1</sup> per year Norg: Organic N in kg.ha<sup>-1</sup> per year P2O5: kg.ha<sup>-1</sup> per year C_N_ratio: C:N ratio
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