Lotus Pedunculatus Research Articles

Addition of Tannin-Containing Legumes to Native Grasslands: Effects on Enteric Methane Emissions, Nitrogen Losses and Animal Performance of Beef Cattle

Extensive cattle production on native grasslands is vital to the sustainability of the South American Pampas, and the inclusion of tannin-containing legumes can increase farm profitability, improve nitrogen (N) use and reduce greenhouse gas (GHG) emissions. This study quantified the effects of adding tannin-containing legumes to native grasslands on enteric methane (CH4) emissions, animal performance and N balance in cattle. A crossover design trial was conducted with 22 beef heifers under two treatments: native grassland (NG) and native grassland with the addition of Lotus uliginosus and L. angustissimus (NG+L). The results showed that forage mass disappearance was similar between treatments; however, 54% of the forage disappearance in the NG+L corresponded with legumes, indicating that the heifers in this treatment consumed a better-quality diet. While individual CH4 emissions were similar between treatments, heifers grazing the NG+L showed a higher average daily gain (ADG) and lower emissions intensity than heifers grazing NGs (0.25 vs. 0.58 g CH4/g ADG, p < 0.05). Additionally, they also ingested 20% more N and were more efficient in its utilization. Incorporating tannin-containing legumes into native grasslands can improve animal productivity and N use efficiency while reducing the intensity of enteric CH4 emissions.

The Effects of Soil Acidity and Aluminium on the Root Systems and Shoot Growth of Lotus pedunculatus and Lupinus polyphyllus.

Lotus pedunculatus (lotus) and Lupinus polyphyllus (Russell lupin) persist in the upland grasslands of New Zealand, where soil acidity and associated aluminium (Al) toxicity impede conventional pasture legumes. This experiment investigated the response of lotus and Russell lupin to soil acidity and Al. The species were sown in 20 cm tall 1.2 L pots of acidic upland soil. A mass of 4.5 or 6.7 g lime (CaCO3)/L was added to either the top or bottom or both soil horizons (0-9 cm and 9-18 cm), resulting in six treatments across six randomised blocks in a glasshouse. The soil pH was 4.4, 4.9, and 5.4; the exchangeable Al concentrations were 24, 2.5, and 1.5 mg/kg for 0, 4.5, and 6.7 g lime/L. At 16 weeks post-sowing, the plants were divided into shoots and roots at 0-9 cm and 9-18 cm. Root morphology, shoot and root dry matter (DM), shoot nitrogen (N), and nodulation were measured. The total plant DM and shoot-to-root DM ratio were higher, and the shoot %N was lower for the lotus plants than the Russell lupin plants for the various lime rates (13.2 vs. 2.9 g plant-1, 5.6 vs. 1.6, and 2.4 vs. 3.3%, p < 0.05). No response to lime in terms of total DM or total root morphology parameters was exhibited in either species (p > 0.05). Root morphology adjustments in response to acidity between soil horizons were not observed. The results indicated that lotus and Russell lupin are tolerant to high soil acidity (pH 4.4-5.4) and exchangeable Al (1.5-24 mg kg-1), highlighting their considerable adaptation to grasslands with acidic soils.

Investigating the efficacy of purified tannin extracts from underutilized temperate forages in reducing enteric methane emissions in vitro

The study investigated how the concentration and composition of purified tannin extracts, at various inclusion rates, affect the ruminal in vitro fermentation parameters. Tannin extracts were isolated from four different forage species: birdsfoot trefoil (Lotus corniculatus), sulla (Hedysarum coronarium), big trefoil (Lotus pedunculatus), and salad burnet (Sanguisorba minor). Plants extracts were purified by Sephadex LH-20 gel chromatography and analyzed by UPLC–ESI–MS/MS. The results showed a large variation among the extracts from different species in terms of tannin composition and structural features. The extracts from salad burnet were dominated by hydrolysable tannins, comprising mainly ellagitannins. The extracts derived from sulla and big trefoil contained predominantly proanthocyanidins (PA), primarily composed of prodelphinidins with high mean degree of polymerisation (mDP). Birdsfoot trefoil extracts comprised procyanidin-rich PAs with low mDP. To determine whether the combined presence of tannins and flavonoid together lead to synergistic or antagonistic effects, the tannin extracts were incubated both with or without rutin at concentrations of 10, 20, and 30 g/kg DM, using a base substrate of perennial ryegrass (Lolium perenne, control). In general, all the tannin extracts decreased methane (CH4) production compared to the control, while no significant effect of rutin was observed on both gas (GP) and CH4 production, neither pure, nor in the simultaneous presence of tannins. The highest CH4 reduction (15%, at 30 g/kg DM) was observed from sulla and big trefoil extracts compared to control, but this was also supplemented with a concomitant reduction in GP (11%) indicating a reduction in feed digestibility. The extracts from birdsfoot trefoil and salad burnet reduced CH4 by up to 12% without significantly reducing GP, indicating the importance of tannin composition on ruminal fermentation.

Responses of Lotus pedunculatus to sulphur fertiliser and defioliation regime in acid soils at low temperature

The potential for Lotus pedunculatus is greatest in acid soils on hill and high country, but requires establishment success, nutrient supply and grazing management. This work investigated the role of sulphur fertiliser and the influence of defoliation regime on production of Lotus in low pH, high aluminium soils. Yields measured in year one were positively related to soil sulphate but not phosphorus. Sulphur, applied at two sites (low or high productivity) at 0, 24 or 78 kg S/ha, failed to alter yield over three years. The site, at 1000 m a.s.l., had base vegetation of unimproved tussock, low soil pH and P (4.8 and 7 respectively) and high Al (30-40 ppm). Yields ranged between 1900 and 3200 kg DM/ha/annum and was reduced when initial plant numbers were lower. Lotus contributed 65- 85% of the herbage for grazing (excluding tussocks). Yields were maximised by defoliation to 2 cm height each season or once in autumn. Establishing high plant numbers increased productivity. Single, late season grazing promoted rhizome development and resulted in the highest yields by year 3. Grazing to 2 or 5 cm residual once a season produced similar herbage, but lower rhizome development.

Plant species, nitrogen status and endophytes are drivers of soil microbial communities in grasslands

Context There is concern that the introduction of ‘novel’ plant germplasm/traits could outpace our capacity to measure and so assess their impacts on soil microbial communities and function. Aim This study aimed to investigate the effects of plant species/functional traits, nitrogen (N) fertilisation and endophyte infection on grassland soil microbial communities within a short time span of 2 years. Methods Two field experiments with monoculture plots were conducted in a common soil. Experiment 1 compared grasses and legumes, using two cultivars of perennial ryegrass (Lolium perenne) that varied in fructan content, along with the legumes white clover (Trifolium repens) and bird’s-foot trefoil (Lotus pedunculatus) that varied in tannin content. Grass treatments received high and low N application levels. Experiment 2 compared the presence/absence of Epichloë strains in ryegrass, tall fescue (Schedonorus phoenix) and meadow fescue (Schedonorus pratensis). Soil microbial communities were analysed by using high-throughput sequencing of DNA isolated from bulk soil cores. Key results Higher abundance of ligninolytic fungi was found in grass soils and pectinolytic fungi in legume soils. Levels of N fertilisation and fructan in ryegrass had only minor effects on soil fungal communities. By contrast, N fertilisation or fixation had a strong effect on bacterial communities, with higher abundance of nitrifiers and denitrifiers in high-N grass soils and in legume soils than in low-N grass soils. Epichloë affected soil microbiota by reducing the abundance of certain fungal phytopathogens, increasing mycorrhizal fungi and reducing N-fixing bacteria. Conclusions Chemical composition of plant cell walls, which differs between grasses and legumes, and presence of Epichloë in grasses were the main drivers of shifts in soil microbial communities. Implications Impacts of farming practices such as mono- or poly-culture, N fertilisation and presence of Epichloë in grasses on soil microbial communities should be considered in pasture management.

Effects of selected condensed tannins on Cryptosporidium parvum growth and proliferation in HCT-8 cell cultures

Infections with Cryptosporidium spp. constitute a substantial public health burden and are responsible for widespread production losses in cattle herds. Reducing disease and shedding of Cryptosporidium spp. oocysts is an important One Health goal. There are very few therapeutic options available to treat cryptosporidiosis. Interest in plant bioactive compounds to mitigate the spread of anthelmintic resistance in ruminants has led to investigation of these phytocompounds against other parasitic taxa. Condensed tannins (CTs) are plant secondary metabolites that have shown potential against nematodes in vitro and in vivo but their applicability to Cryptosporidium spp. is comparatively under-explored. Cryptosporidium parvum infected human ileocecal colorectal adenocarcinoma (HCT)-8 cell cultures were treated with escalating doses of highly purified and well-characterized CTs from five plant species, big trefoil (Lotus pedunculatus), black currant (Ribes nigrum), sainfoin (Onobrychis viciifolia), white clover (Trifolium repens) and grapeseed (Vitis vinifera) for 44 h. Quantitative-PCR (qPCR) analysis revealed that none of the CTs examined demonstrated inhibitory potential against the parasite. Substantial inhibition of C. parvum by paromomycin was observed in positive controls in all assays (76.94–90.72% inhibition), proving the validity of the assay. Despite the lack of inhibition, these results represent an important step towards identifying alternative treatment options against this parasite.

Long-term effects of artificial nighttime lighting and trophic complexity on plant biomass and foliar carbon and nitrogen in a grassland community.

The introduction of artificial nighttime lighting due to human settlements and transport networks is increasingly altering the timing, intensity, and spectra of natural light regimes worldwide. Much of the research on the impacts of nighttime light pollution on organisms has focused on animal species. Little is known about the impacts of daylength extension due to outdoor lighting technologies on wild plant communities, despite the fact that plant growth and development are under photoperiodic control. In a five‐year field experiment, artificial ecosystems (“mesocosms”) of grassland communities both alone or in combination with invertebrate herbivores and predators were exposed to light treatments that simulated street lighting technologies (low‐pressure sodium, and light‐emitting diode [LED]‐based white lighting), at ground‐level illuminance. Most of the plant species in the mesocosms did not exhibit changes in biomass accumulation after 5 years of exposure to the light treatments. However, the white LED treatment had a significant negative effect on biomass production in the herbaceous species Lotus pedunculatus. Likewise, the interaction between the white LED treatment and the presence of herbivores significantly reduced the mean shoot/root ratio of the grass species Holcus lanatus. Artificial nighttime lighting had no effect on the foliar carbon or nitrogen in most of the grassland species. Nevertheless, the white LED treatment significantly increased the leaf nitrogen content in Lotus corniculatus in the presence of herbivores. Long‐term exposure to artificial light at night had no general effects on plant biomass responses in experimental grassland communities. However, species‐specific and negative effects of cool white LED lighting at ground‐level illuminance on biomass production and allocation in mixed plant communities are suggested by our findings. Further studies on the impacts of light pollution on biomass accumulation in plant communities are required as these effects could be mediated by different factors, including herbivory, competition, and soil nutrient availability.

Linking metabolites in eight bioactive forage species to their in vitro methane reduction potential across several cultivars and harvests

An in vitro Hohenheim gas test was conducted to analyze the fermentation end-products from 17 cultivars of eight polyphenol containing forage species. The polyphenol composition and proanthocyanidin (PA) structural features of all the cultivars were analyzed with UPLC-MS/MS in leaves of vegetative or generative plants. The samples were incubated with and without polyethylene glycol (PEG, a tannin-binding agent) to separate the tannin-effect on methane (CH4, ml/200 mg DM) production from that of forage quality. Sulla and big trefoil, two particularly PA rich species, were found to have the highest CH4 reduction potential of up to 47% when compared to the samples without PEG. However, concomitant reduction in gas production (GP, ml/200 mg DM) of up to 44% was also observed. An increase in both GP and CH4 production under PEG treatments, confirms the role of tannins in CH4 reduction. Moreover, PA structural features and concentration were found to be an important source of variation for CH4 production from PA containing species. Despite having low polyphenol concentrations, chicory and plantain were found to reduce CH4 production without reducing GP. Additionally, interspecies variability was found to be higher than intraspecies variability, and these results were consistent across growth stages, indicating the findings’ representativeness.

Assessing the Potential of Diverse Forage Mixtures to Reduce Enteric Methane Emissions In Vitro.

Simple SummaryChanges in agriculture towards simpler and more intensive systems have contributed to increased environmental problems. In temperate sown grasslands, this has resulted in ryegrass dominance, and forage legume use is limited mainly to three species: red clover, white clover and lucerne. Other dicot forages, such as Lotus pedunculatus and Sanguisorba minor, are of interest as they contain plant specialized metabolites (PSM), especially tannins, potentially reducing methane from ruminants, an important source of agricultural greenhouse gas emissions. In an in vitro study, we compared binary mixtures of perennial ryegrass with one of eight dicot species, including PSM-rich species in different proportions, to assess their potential to reduce methane production. An additional aim was to determine whether moderate additions of these forage species can be sufficient to reduce methane formation or whether the relationship is linearly dose-dependent. Results show that all dicot species studied, including the non-tannin-containing species, reduced methane production. While all plant species rich in PSM reduced methane production, they also decreased digestibility. Additionally, they did not persist in the pasture into the second year. The lowest methane emissions per digestible forage unit were obtained with chicory (Cichorium intybus), a promising forage herb with both agronomic and bioactive potential.Methane emissions from ruminants are a major contributor to agricultural greenhouse gas emissions. Thus, eight different forage species were combined in binary mixtures with Lolium perenne in increasing proportions, in vitro, to determine their methane reduction potential in ruminants. Species were sampled in two consecutive years where possible. The aims were: a) to determine if mixtures with specific forages, particularly those rich in plant specialized metabolites (PSM), can reduce methane emissions compared to ryegrass monocultures, b) to identify whether there is a linear-dose effect relationship in methane emissions from the legume or herb addition, and c) whether these effects are maintained across sampling years. Results showed that all dicot species studied, including the non-tannin-containing species, reduced methane production. The tannin-rich species, Sanguisorba minor and Lotus pedunculatus, showed the greatest methane reduction potential of up to 33%. Due to concomitant reductions in the forage digestibility, Cichorium intybus yielded the lowest methane emissions per digestible forage unit. Contrary to total gas production, methane production was less predictable, with a tendency for the lowest methane production being obtained with a 67.5% share of the legume or herb partner species. Thus, linear increments in the partner species share did not result in linear changes in methane concentration. The methane reduction potential differed across sampling years, but the species ranking in methane concentration was stable.

Methane emissions from lambs fed kikuyu hay alone or mixtured with lotus hay

Dietaryinclusionof contain­tanninlegumes mayreduce enteric methane emissioninruminants. Toevaluate methane emissions fromsheep fed with a kikuyu grass (Cenchrus clandestinus) diet partially substitutedwith lotus(Lotus uliginosus), twelve growing lambs, with 23 ± 2 kg average liveweight, were assigned randomly totwo treatments and with three measurement periods in a switchover design. Treatments consisted of 100 %kikuyuhay or 70 %kikuyu hay: 30 %lotus hay and with 6 lambs per treatment. Each of three periods lasted 20 d, where thefirst 15 d were for acclimatization and the last 5 d for measurements. Lambs were placed in metabolic cages and fedonce a day (8 AM) at 90 %of their voluntary feedintake, with free access to drinking water. Feedintake, fecalproductionandfeeddigestibility were determinedat eachperiod. Methane productionwas measuredfor eachtreatment group of 6 lambs using the poly­tunnel technique. Legume addition reduced total methane production(27.6vs.23.1 g animal­1; p &lt; 0.01), methane production per dry matter intake (DMI) (18.8 vs. 12.2 g kg­1DMI; p &lt;0.01), methane production per digestible OM(DOM) (36.1vs. 23.4 g kg­1DOM; p &lt; 0.01) and methane productionper digestible NDF (DNDF) (43.5vs. 34.0 g kg­1DNDF; p &lt; 0.01). In conclusion, lotus inclusion in pasture systemscould be a suitable legume to reduce methane emissions in grazing systems.

Productivity and nutritional value of 20 species of perennial legumes in a low‐rainfall Mediterranean‐type environment in southern Australia

AbstractIn the rain‐fed mixed‐farming systems of southern Australia, the consistent supply of high‐value forage is limited by a range of climatic, edaphic and systems constraints. Over 2 years, we compared biomass production and nutritional value of 30 accessions of perennial legumes, and predicted intake, grazing days and growth of ewes and lambs. There was significant variation in nutritional value and biomass production between and within species. Lucerne (Medicago sativa) and sulla (Hedysarum coronarium) produced the greatest amount of biomass and energy. There was variability among accessions in digestibility (DMD), acid detergent fibre (ADF) and crude protein, and the rate of change in these traits as plants matured. Trifolium species had the highest DMD across all growth stages. Hairy canary clover (Dorycnium hirsutum), erect canary clover (Dorycnium rectum), greater birdsfoot trefoil (Lotus uliginosus), Australian trefoil (Lotus australis) and running postman (Kennedia prostrata) had energy levels that would not maintain liveweight of mature sheep. In the second year, species differed in response to harvesting treatments. Lucerne and sainfoin (Onobrychis viciifolia) were more productive under a frequent cutting regime. Accessions of white clover (Trifolium repens), red clover (Trifolium pratense), alsike clover (Trifolium hybridum), cullen (Cullen australasicum), strawberry clover (Trifolium fragiferum), sainfoin and birdsfoot trefoil (Lotus corniculatus) showed some promise, while Tedera (Bituminaria bituminosa) and milkvetch (Astragalus cicer) performed poorly under the experimental conditions. We conclude by discussing additional agronomic and nutritional factors that need consideration when developing novel perennial legumes for mixed‐farming systems in the context of a changing climate.

24 Condensed tannins: structure, activity and characterization

Abstract As a class of plant polyphenolic compounds contained in some forages [i.e., sainfoin (Onobrychis viciifolia Scop.), big trefoil (Lotus pedunculatus Cav.), birdsfoot trefoil (Lotus corniculatus L.)], condensed tannins (CTs) exhibit a variety of biological effects on ruminants. The potential positive impact of CTs on the agricultural industry stems from their ability to modulate proteolysis during forage conservation and ruminal digestion, to prevent bloat, to reduce intestinal parasite burdens, and to abate methane and ammonia emissions from ruminants. How CTs exert these effects on ruminants focuses on the interaction of CTs with proteins. The structure-activity relationship in CT–protein interaction is not well understood but is known to be dependent on the structure and properties of both the CT and the protein. The objectives of this presentation are fivefold. First, examples of the structural diversity of CTs will be provided to enable the audience members to appreciate that not all CTs are the same. Second, examples of how CTs structural diversity affects their interaction with the protein, which in turn, dictates the biological response from the animal will be discussed. Third, the presentation will outline hurdles in obtaining highly pure and well-characterized CTs from natural sources for use in CT structural analysis and in vitro experiments. This will be followed by brief descriptions of improved and emerging techniques for CT analysis and, finally, the presentation concludes with questions to address in future investigations and a list of recommendations for CT researchers to follow.

Can Lotus pedunculatus over-sowing in low-fertility tussock country increase farm resilience?

The range of legumes to boost farm productivity in low-fertility hill country are limited. Lotus pedunculatus (Lotus) provides an option when soil pH is below 5 but is intolerant to severe and regular grazing. However, itcan be used at sites that are only grazed occasionally during spring summer and autumn. Oversowing of Lotus pedunculatus has been used to improve 17% of the total area of Avenel Station, Millers Flat. Measurements of pasture growth and animal production were collected over three years post-sowing, indicating a doubling of pasture production from the native form while providing liveweight gains of 0.135 and 0.75 kg/d in lambs and yearling cattle respectively. Systems modelling was used to re-examine the current farm-system configuration to test the possible outcomes from implementing further enterprises that may capture the increased late spring and summer production effectively. Reducing ewe numbers and concentrating on improving lamb survival were the most cost-effective and environmentally effective options. Buying in further finishing lambs to use the summer surplus also improved profitability. Increasing cow numbers was similarly profitable. Increasing ewe numbers also required an increase in winter feed supply and was the least profitable way to capture the benefits. Testing using climatic extreme scenarios demonstrated that the oversowing of Lotus on the higher, wetter tussock country increased the resilience of the farm by providing a buffer during dry summers.

Species selection and assessment of eco‐engineering effects of seedlings for biogeomorphological landscape experiments

AbstractLandscape experiments of fluvial environments such as rivers and deltas are often conducted with live seedlings to investigate effects of biogeomorphological interactions on morphology and stratigraphy. However, such experiments have been limited to a single species, usually alfalfa (Medicago sativa), whereas important environments in nature have many different vegetation types and eco‐engineering effects. Landscape experimentation would therefore benefit from a larger choice of tested plant species. For the purpose of experimental design our objective was to identify fast‐germinating and fast‐growing species and determine their sensitivity to flow conditions during and after settling, their maximum growth, hydraulic resistance and added bank strength. We tested germination time and seedling growth rate of 18 candidate species with readily available seeds that are fast growing and occur at waterlines, plus Medicago sativa as a control. We selected five species that germinate and develop within days and measured properties and eco‐engineering effects depending on plant age and density, targeting typical experimental conditions of 0–0.3 m/s flow velocity and 0–30 mm water depth. Tested eco‐engineering effects include bank strength and flow resistance. We found that Rumex hydrolapathum can represent riparian trees. The much smaller Veronica beccabunga and Lotus pedunculatus can represent grass and saltmarsh species as they grow in dense patches with high flow resistance but are readily erodible. Sorghum bicolor grows into tall, straight shoots, which add significantly to bank strength, but adds little flow resistance and may represent sparse hardwood trees. Medicago sativa also grows densely under water, suggesting a use for mangroves and perhaps peat. In stronger and deeper flows the application of all species changes accordingly. These species can now be used in a range of landscape experiments to investigate combined effects on living landscape patterns and possible facilitation between species. The testing and treatment methodology can be applied to new species and other laboratory conditions. © 2019 The Authors. Earth Surface Processes and Landforms Published by John Wiley &amp; Sons Ltd. © 2019 The Authors Earth Surface Processes and Landforms Published by John Wiley &amp; Sons Ltd © 2019 The Authors Earth Surface Processes and Landforms Published by John Wiley &amp; Sons Ltd.

Effect of Condensed Tannins in Lotus uliginosus cv. E-Tanin and/or Extracts of Quebracho and Chestnut in Carcass and Lamb Meat

The effect of condensed tannins (CT) in the pasture (Lotus uliginosus cv. E-Tanin) and/or quebracho and chestnut extracts on lamb carcass and meat quality was evaluated. Two experiments were conducted in consecutive years with 60 crossbreed lambs each year. The animals were stratified by live weight (LW) and randomly assigned to the treatments: white clover grazing (WC); WC plus concentrate supplementation (WC + C); WC plus concentrate supplementation and quebracho and chestnut extract (WC + C + CT); Lotus uliginosus grazing (E-Tanin); E-Tanin plus concentrate supplementation (E-Tanin + C); E-Tanin plus concentrate supplementation plus quebracho and chestnut extract (E-Tanin + C + CT); with two repetitions each year. Pasture type affected loin weight, which was greater (P &lt; 0.05) in lambs grazing WC than E-Tanin. Meat color redness (a*) was greater (P &lt; 0.05) in supplemented treatments with C (with or without CT) than those exclusively grazing. Intramuscular fat content was not affected (P &gt; 0.05) by the pasture, the supplementation, or the year. The proportion of dihomo-gamma-linolenic acid (C20:3-n6) was greater (P &lt; 0.05) in lambs grazing E-Tanin compared to WC. Supplementation affected (P &lt; 0.05) the saturated (SFA) and monounsaturated fatty acids (MUFA) proportions, and the omega 6:omega 3 ratio. Estimation of dry matter intake under grazing conditions would allow a better understanding of the effect of CT on carcass and meat quality.

Activity, Purification, and Analysis of Condensed Tannins: Current State of Affairs and Future Endeavors

As a class of plant polyphenolic compounds contained in some forages (i.e., sainfoin [Onobrychis viciifolia Scop.], big trefoil [Lotus pedunculatus Cav.], birdsfoot trefoil [Lotus corniculatus L.]), condensed tannins (CTs), also referred to as proanthocyanidins (PAs or PACs), exhibit a variety of biological effects on ruminants. The potential positive impact of CTs on the agricultural industry stems from their ability to modulate proteolysis during forage conservation and ruminal digestion, to prevent bloat, to reduce intestinal parasite burdens, to abate methane and ammonia emissions from ruminants, and to inhibit the activity of soil‐nitrifying bacteria. How CTs exert these effects on ruminants focuses on the interaction of CTs with proteins. The structure‐activity relationship in CT–protein interaction is not well understood but is known to be dependent on the structure and properties of both the CT and the protein. The purpose of this perspective is fivefold. First, we provide the reader with a better understanding of the structural diversity of CTs present in plant material and enable the reader to appreciate that not all CTs are the same. Second, we provide examples of how CT structural diversity affects the interaction of CTs with the protein, which, in turn, dictates the biological response from the animal. Third, we describe the hurdles in obtaining highly pure and well‐characterized CTs from natural sources for use in studies to attempt to elucidate how CTs impart each biological effect. We then describe improved and emerging techniques for CT analysis and, finally, we conclude this perspective with questions to address in future investigations and forward a list of recommendations for CT researchers to follow.

Dry matter yield of six perennial legume species in response to lime over 3 years at Glenmore Station, Mackenzie Basin

The production and persistence of legumes which may improve South Island high-country pasture in the Mackenzie Basin were studied. The initial soil test results confirmed toxic levels of aluminium (9 mg/kg, 0-150 mm soil depth) that has prevented the development of large areas of land traditionally grazed by merino sheep and beef cattle. A 3-year field experiment was direct-drilled in December 2012 with five rates (0, 0.5, 1, 2, and 4 t/ha) of surface applied lime and six legumes (lucerne, ‘Russell’ lupin, Caucasian clover, white clover, balansa, and Lotus pedunculatus) in three replicates. In Year 1, balansa clover and Russell lupin were the highest yielding legumes (7 t DM/ha), regardless of lime application rates. In Year 2 ‘Russell’ lupin was the highest (10.4 t DM/ha). Caucasian clover produced 4.6 t DM/ha and white clover and lucerne around 4.0 t DM/ha. Balansa clover (1.4 t DM/ha) and Lotus pedunculatus (2.5 t DM/ha) were the lowest yielding and they did not recover or persist after the initial grazing by merino sheep. These results confirmed that ‘Russell’ lupin and Caucasian clover thrived without the application of lime in this acidic soil with high Al levels.

Phytososological analysis of hydro ecosystems of the NDP «North Podillya»

It is substantiated that the territory of the NNP «Pivnichne Podillia» is unique in the phytososological meaning of the ecosystem — three species of plants (Cochlearia pyrenaica DC., Sesleria caerulea (L.) Ard., Spiranthes amoena (M. Bieb.) Spreng.) occur in Ukraine only here, the location of which was confirmed by us in 2017 new NNP ecotops of the Red Data Book of Ukraine were marked: Utricularia intermedia Hayne, U. minor L., Salix lapponum L., Dactylorhiza maculata (L.) Soo, D. majalis (Rchb.) PF Hunt et Summerhayes. A part of the species of the Red Data Book of Ukraine, found in literary sources, have been identified. The list of regionally rare plants of Lviv region is analyzed. There are 10 phytogroups of the Green Data Book of Ukraine. Rare wetland types of natural environments of the Bern Convention are presented. After further research, we recommend the inclusion of the following vulnerable, rare, endangered species of flora of wetlands and peatlands of the North Podillia: Drosera rotundifolia L., Lathyrus palustris L., Ophioglossum vulgatum L., Veratrum lobelianum Bernh., Lotus uliginosus L., Eriophorum gracile Koch, Parnassia palustris L., Triglochin palustre L., Blysmus compressus (L.) Panz. ex Link., to the fourth edition of the Red Data Book of Ukraine.

Physiological and biochemical responses to water deficit in Lotus uliginosus × L. corniculatus hybrids

Lotus uliginosus (greater lotus, GL) and L. corniculatus (birdsfoot trefoil, BT) are species markedly different in their genetics, morphology and environmental adaptation. We evaluated the hybrids and parental species under differential conditions: in vitro culture, growth chamber and open field environments. The experimental evaluation included biochemical, physiological, developmental and productive parameters. Parental species exhibited significant differences in root growth under different osmotic potentials generated by polyethylene glycol, and hybrids exhibited variability in their response compared with their parents. Plants grown in pots and subjected to drought exhibited differences in biochemical parameters. Proline accumulation and oxidative damage measured by lipid peroxidation were higher in birdsfoot trefoil than any other genotype, and the lowest values were observed in greater lotus. The total phenolic content in shoot ranged from 147 to 279 mg gallic acid equivalents 100 g–1 dry mater. With regard to antioxidant capacity estimated by 2,2-diphenyl-1-picrylhydrazyl, greater lotus had the highest value and birdsfoot trefoil the lowest, at almost one-third, whereas hybrids had intermediate values. Under stress conditions, water-use efficiency showed significant differences between both parental species. In the field, under either irrigation or drought, the dry matter accumulated by the hybrids was higher than that of the parents. Hybrids express recombination of features, making them an interesting material to continue evaluation.

Detection of habitats colonized by Omphiscola glabra (Gastropoda: Lymnaeidae) on acid soils using indicator plants

The lymnaeid Omphiscola glabra is known to be an intermediate host in the life cycle of Fasciola hepatica and knowing its distribution in the field is important to determine that of the parasite. Looking at indicator plants has been proposed by several authors as one of the most practical ways of carrying out a malacological survey in pastures of a farm. Field investigations on 361 French farms breeding cattle or sheep on acid soils were thus carried out for 36 years in March or April to list indicator plant species in seven types of habitats colonized by O. glabra , and determine the characteristics of their habitats (area, snail density) for each indicator plant. Six plant species: Agrostis stolonifera , Dactylis glomerata , Glyceria fluitans , Juncus acutiflorus , Juncus effusus and Lotus uliginosus , were examined in 3137 snail habitats. J. effusus was the best indicator species to detect snail populations in five habitat types, while J. acutiflorus was the best indicator for snail populations living in spring heads. The other four plant species characterized only a few populations. Snail habitats detected with J. acutiflorus , J. effusus or L. uliginosus were significantly larger in area, while the density of overwintering snails was significantly lower in habitats with G. fluitans . These initial results suggest that J. effusus is a good indicator for the detection of O. glabra in the acid soils of central France. However, additional studies are required to test its limitations in the field.