### Recent Submissions

• Journal Article

#### Consumers’ Evaluation of Stockfree-Organic Agriculture—A Segmentation Approach ﻿

Sustainability 2020; 12(10) p.1-19: Art. 4230
Recently, more and more research has been conducted on what sustainable nutrition could look like. Stockfree-organic agriculture is one possible approach but a relatively new and unstudied cultivation method. In addition to organic agriculture, it excludes any animal by-products during the whole cultivation process. Among the consumers of organic food are especially many vegetarians and vegans. To attract this target group, first farms in Europe have started to follow the stockfree-organic agriculture principles. As it is important to know the consumers’ point of view on new developments in agriculture at an early stage of the diffusion process, this study deals with consumers’ evaluation of stockfree-organic agriculture to draw conclusions about a possible market potential. This is especially important for stockfree-organic farmers, as well as for organic farmers who are considering converting their cultivation method, and for retailers who wonder whether it is worthwhile to offer these products. The data was collected in 2019 by means of an online survey. The sample consisted of 500 German respondents. Principal component and cluster analyses were used to identify consumer segments according to their attitudes towards the acceptance, advantages, and disadvantages of stockfree-organic agriculture. Additionally, the different segments were compared with each other according to various attitudes and eating behaviours. Overall, animal welfare considerations and environmental aspects were of particular importance to consumers. Animal usage was clearly rejected by one segment, which contained 24% of the sample. Nearly all vegetarians and all vegans supported stockfree-organic agriculture, whereas heavy meat consumers tended to refuse the support of stockfree-organic agriculture. The supporting group valuing high animal welfare and health concerns was much larger than the current status of this niche segment would suggest. This could be a major challenge for the agricultural sector in the long term, but could also include opportunities for greater sustainability.
• Journal Article

#### Growth Performance of Local Chicken Breeds, a High-Performance Genotype and Their Crosses Fed with Regional Faba Beans to Replace Soy ﻿

Animals 2020; 10(4) p.1-19: Art. 702
The faba bean (Vicia faba L.) is a native protein crop and considered a promising alternative to soybeans. Due to its anti-nutritive substances such as vicin and convicin (VC) its use in animal nutrition has been restricted. In the present study, two consecutive experiments were conducted to analyse the effects of feeding 20% faba beans, which differ in their VC content on fattening performance and slaughter traits of different chicken genotypes. In a first trial, purebred male chickens of the local breeds Bresse Gauloise and Vorwerkhuhn as well as of a high-performance White Rock line were tested. In a second trial, crossbreds of them were evaluated: Vorwerkhuhn $\times$ Bresse Gauloise, Vorwerkhuhn $\times$ White Rock, Bresse Gauloise $\times$ White Rock. Daily weight gain and feed intake were recorded until slaughter at approximately 2100 g. At slaughter the final live weight, carcass yield and the weights of the valuable parts (breasts and legs) were measured. For the genotypes studied, no adverse or undesirable effects of both VC-rich and VC-poor faba beans in the feedstuff were detected regarding body weight development, carcass quality, and fattening parameters. Furthermore, there was no indication that the birds’ health was impaired.
• Journal Article

#### Effect of Alternative Protein Feeds on the Content of Selected Endogenous Bioactive and Flavour-Related Compounds in Chicken Breast Meat ﻿

Foods 2020; 9(4) p.1-8: Art. 392
Currently, soybean meal constitutes the main protein source for poultry production. However, the environmental and social issues related to soybean production are calling for more sustainable protein sources that can offset soybean requirements in animal production. Hermetia illucens larvae and the microalga spirulina (Arthrospira platensis) have proven to be effective alternatives to soybean meal for poultry production. In this study, the effect of 100% replacement of soy with partially defatted Hermetia illucens larvae and spirulina on the contents of selected endogenous bioactive (anserine, creatine and carnosine) and flavour-related (inosine and inosine-5´-monophosphate, IMP) compounds in chicken breast meat was evaluated. The results showed that the spirulina-based diet lowered the levels of anserine, carnosine and creatine compared to the control diet (3.3 vs. 4.1 mg/g, 0.15 vs. 0.72 mg/g and 1.49 vs. 2.49 mg/g, respectively) while IMP levels tended to be higher in spirulina-fed samples. Compared to the control group, Hermetia illucens-fed samples showed a lower content of bioactive peptides (anserine: 3.6 vs. 4.1 mg/g; carnosine: 0.39 vs. 0.72 mg/g; creatine: 2.03 vs. 2.49 mg/g), albeit to a lesser extent than the spirulina treatment group.
• Journal Article

#### Identification of Age-Specific and Common Key Regulatory Mechanisms Governing Eggshell Strength in Chicken Using Random Forests ﻿

Genes 2020; 11(4) p.1-18: Art. 464
In today’s chicken egg industry, maintaining the strength of eggshells in longer laying cycles is pivotal for improving the persistency of egg laying. Eggshell development and mineralization underlie a complex regulatory interplay of various proteins and signaling cascades involving multiple organ systems. Understanding the regulatory mechanisms influencing this dynamic trait over time is imperative, yet scarce. To investigate the temporal changes in the signaling cascades, we considered eggshell strength at two different time points during the egg production cycle and studied the genotype–phenotype associations by employing the Random Forests algorithm on chicken genotypic data. For the analysis of corresponding genes, we adopted a well established systems biology approach to delineate gene regulatory pathways and master regulators underlying this important trait. Our results indicate that, while some of the master regulators ($Slc22a1$ and $Sox11$) and pathways are common at different laying stages of chicken, others (e.g., $Scn11a$, $St8sia2$, or the TGF-$\beta$ pathway) represent age-specific functions. Overall, our results provide: (i) significant insights into age-specific and common molecular mechanisms underlying the regulation of eggshell strength; and (ii) new breeding targets to improve the eggshell quality during the later stages of the chicken production cycle.
• Journal Article

#### Improved Protocol for DNA Extraction from Subsoils Using Phosphate Lysis Buffer ﻿

Microorganisms 2020; 8(4) p.1-16: Art. 532
As our understanding of soil biology deepens, there is a growing demand for investigations addressing microbial processes in the earth beneath the topsoil layer, called subsoil. High clay content in subsoils often hinders the recovery of suffcient quantities of DNA as clay particles bind nucleic acids. Here, an effcient and reproducible DNA extraction method for 200 mg dried soil based on sodium dodecyl sulfate (SDS) lysis in the presence of phosphate buffer has been developed. The extraction protocol was optimized by quantifying bacterial 16S and fungal 18S rRNA genes amplified from extracts obtained by different combinations of lysis methods and phosphate buffer washes. The combination of one minute of bead beating, followed by ten min incubation at 65°C in the presence of 1 M phosphate buffer with 0.5% SDS, was found to produce the best results. The optimized protocol was compared with a commonly used cetyltrimethylammonium bromide (CTAB) method, using Phaeozem soil collected from 60 cm depth at a conventional agricultural field and validated on five subsoils. The reproducibility and robustness of the protocol was corroborated by an interlaboratory comparison. The DNA extraction protocol offers a reproducible and cost-effective tool for DNA-based studies of subsoil biology.
• Journal Article

#### Effect of Grazing System on Grassland Plant Species Richness and Vegetation Characteristics: Comparing Horse and Cattle Grazing ﻿

Sustainability 2020; 12(8) p.1-17: Art. 3300
Horses are of increasing relevance in agriculturally managed grasslands across Europe. There is concern to what extent grazing with horses is a sustainable grassland management practice. The effect of longer-term horse grazing on the vegetation characteristics of grasslands has received little attention, especially in comparison to grazing cattle. Our study analyses the relative importance of grazing system (grazer species and regime) and grassland management for vegetation characteristics in grasslands as indicator for sustainable management. We monitored grassland vegetation in western central Germany and compared paddocks grazed by horses under two different regimes, continuous (HC) vs. rotational (HR), to paddocks grazed by cattle (C) under similar trophic site conditions. We observed more plant species and more High Nature Value indicator species on HC compared to C. The vegetation of C was more grazing tolerant and had higher forage value than HC. Regardless of the grazing regime, the competitive component was lower, the stress-tolerant component higher and the floristic contrast between patch-types stronger on HC and HR paddocks compared to C. Species richness was strongly influenced by the extent of the floristic contrast. Our results emphasize the potential of horse grazing for biodiversity in agriculturally managed grasslands.
• Journal Article

#### Acceptance and Feasibility of a Guideline for the Animal Welfare Assessment of Fattening Pigs from Farmers’ Point of View ﻿

Animals 2020; 10(4) p.1-14: Art. 711
The welfare of farm animals is being increasingly discussed in society and politics. To evaluate animal welfare, indicator systems are often used. Such a system has been developed by the German Association for Technology and Structures in Agriculture and suggested in the publication “Animal Welfare Indicators: Practical Guide—Pigs”. The association’s aim is to provide farmers with a useful method for recording the welfare of pigs. Crucial for the acceptance of the guide by farmers is a high degree of feasibility of the recommended indicators as well as the proposed methods for their recording. To evaluate this, 40 farmers keeping fattening pigs were interviewed. The guided semi-structured interview was conducted on the farms after the farmers evaluated the welfare of their fattening pigs according to the guide. The results are: Apart from the indicators faecal soiling and tail length, all the other eleven indicators are accepted for the assessment of fattening pig welfare by a majority of the interviewed farmers (between 57.5% and 90% acceptance per indicator). Furthermore, the feasibility of the individual indicators was assessed as being positive. The relationship between time expenditure and benefit was rated on a five-point scale at an average of 3.1 (medium), which clearly shows that there is a need for further development of this guide. Some possible changes with a potential for improvement could be identified; for example, the aggregation of the results after the collection of the individual indicators to an overall result that can be compared and interpreted.
• Journal Article

#### Tail Lesions and Losses of Docked and Undocked Pigs in Different Farrowing and Rearing Systems ﻿

Agriculture 2020; 10(4) p.1-11: Art. 130
This study aimed to investigate the effects of farrowing and rearing systems on tail lesions and losses of docked and undocked pigs. Pigs from three farrowing systems: Conventional farrowing crate (FC), free farrowing (FF) and group housing of lactating sows (GH) were randomly allocated to different rearing systems: A conventional system (CONV), where the pigs were regrouped and transferred to conventional finishing pens at ten weeks of age or a wean-to-finish (W-F) system, where the pigs remained in their pens until slaughter with higher space allowance during rearing. Weekly, tail lesions and losses were assessed individually. The incidence of tail lesions was higher in undocked CONV pigs compared to undocked W-F pigs (maximum: CONV 58.01%, W-F 41.16%). The rearing system had a significant effect on tail losses at the end of finishing (CONV 67.63%,W-F 38.2%). The significant effect of the rearing system might be explained by higher space allowance during rearing and reduced regrouping stress forW-F pigs. In conclusion, farrowing systems showed no effects, but the W-F rearing system reduces the frequency of tail lesions and losses; the curves of tail lesions increased slower and stayed on a lower level, which resulted in lower losses as well.
• Journal Article

#### Performance of Modern Varieties of Festuca arundinacea and Phleum pratense as an Alternative to Lolium perenne in Intensively Managed Sown Grasslands ﻿

Agronomy 2020; 10(4) p.1-13: Art. 540
In future, grass swards need to be adapted to climate change and interactions of management and site are becoming more important. The persistence of Lolium perenne on peatland or during dry periods is limited and alternative forage species are required. We tested the performance of a modern variety of Festuca arundinacea and Phleum pratense as an alternative to Lolium perenne on clay, peat, and sandy soils. Each of these grasses was sown as main species in mixture with Poa pratensis and Trifolium repens and the mixtures were subjected to different frequencies of defoliation. Differences in yield proportions in the third year were significantly influenced by main species, site and their interaction. Remaining mass proportions of main species after three years were smallest on peat; on all sites Festuca arundinacea showed the highest persistence and largest yield, followed by Lolium perenne. Mass proportions of Phleum pratense were small on peat soils and Phleum had been replaced there by Holcus lanatus, and by Lolium perenne and Poa pratensis on the clay and sandy soils. We conclude that the choice of grass species in mixtures is a management tool to control stability and productivity of grass swards under specific site conditions.
• Journal Article

#### High-Resolution Melting (HRM) Curve Assay for the Identification of Eight Fusarium Species Causing Ear Rot in Maize ﻿

Pathogens 2020; 9(4) p.1-13: Art. 270
Maize plants are often infected with fungal pathogens of the genus Fusarium. Taxonomic characterization of these species bymicroscopic examination of pure cultures or assignment to mating populations is time-consuming and requires specific expertise. Reliable taxonomic assignment may be strengthened by the analysis of DNA sequences. Species-specific PCR assays are available for most Fusarium pathogens, but the number of species that infect maize increases the labor and costs required for analysis. In this work, a diagnostic assay for major Fusarium pathogens of maize based on the analysis of melting curves of PCR amplicons was established. Short segments of genes RPB2 and TEF-1$\alpha$, which have been widely used in molecular taxonomy of Fusarium, were amplified with universal primers in a real-time thermocycler and high-resolution melting (HRM) curves of the products were recorded. Among major Fusarium pathogens of maize ears, F. cerealis, F. culmorum, F. graminearum, F. equiseti, F. poae, F. temperatum, F. tricinctum, and F. verticillioides, all species except for the pair F. culmorum/F. graminearum could be distinguished by HRM analysis of a 304 bp segment of the RPB2 gene. The latter two species could be differentiated by HRM analysis of a 247 bp segment of the TEF-1$\alpha$gene. The assay was validated with DNA extracted from pure cultures of fungal strains, successfully applied to total DNA extracted from infected maize ears and also to fungal mycelium that was added directly to the PCR master mix (“colony PCR”). HRM analysis thus offers a cost-efficient method suitable for the diagnosis of multiple fungal pathogens.
• Journal Article

#### Photosynthetic Cost Associated With Induced Defense to Plasmopara viticola in Grapevine ﻿

Frontiers in Plant Science 2020; 11 p.1-14: Art. 235
Downy mildew caused by Plasmopara viticola is one of the most destructive diseases of Vitis vinifera worldwide. Grapevine breeding programs have introgressed P. viticolaresistant traits into cultivated V. vinifera genotypes and launched interspecific hybrids with resistance against downy mildew. In general, pathogen infection affects primary metabolism, reduces plant growth and development and modifies the secondary metabolism toward defense responses, which are costly in terms of carbon production and utilization. The objective of this work was to evaluate the photosynthesis impairment by inducible defenses at the leaf level in V. vinifera cultivars resistant to P. viticola. Photosynthetic limitations imposed by P. viticola in susceptible and resistant grapevine cultivars were evaluated. Histochemical localization of hydrogen peroxide and superoxide and the activity of ascorbate peroxidase were assessed. Measurements of leaf gas exchange, chlorophyll fluorescence and the response of leaf CO$_2$ assimilation to increasing air CO$_2$ concentrations were taken, and photosynthetic limitations determined in cultivars Solaris (resistant) and Riesling (susceptible). The net photosynthetic rates were reduced (-25%) in inoculated Solaris plants even before the appearance of cell death-like hypersensitive reactions (“HR”). One day after “HR” visualization, the net photosynthetic rate of Solaris was reduced by 57% compared with healthy plants. A similar pattern was noticed in resistant Cabernet Blanc and Phoenix plants. While the susceptible cultivars did not show any variation in leaf gas exchange before the appearance of visual symptoms, drastic reductions in net photosynthetic rate and stomatal conductance were found in diseased plants 12 days after inoculation. Decreases in the maximum Rubisco carboxylation rate and photochemical impairment were noticed in Riesling after inoculation with P. viticola, which were not found in Solaris. Damage to the photochemical reactions of photosynthesis was likely associated with the oxidative burst found in resistant cultivars within the first 24 h after inoculation. Both chlorophyll degradation and stomatal closure were also noticed in the incompatible interaction. Taken together, our data clearly revealed that the defense response against P. viticola causes a photosynthetic cost to grapevines, which is not reversible even 12 days after the pathogen infection.
• Journal Article

#### Analyzing the Dietary Diary of Bumble Bee ﻿

Frontiers in Plant Science 2020; 11 p.1-9: Art. 287
Bumble bees are important crop pollinators and provide important pollination services to their respective ecosystems. Their pollen diet and thus food preferences can be characterized through nucleic acid sequence analysis. We present ITS2 amplicon sequence data from pollen collected by bumble bees. The pollen was collected from six different bumble bee colonies that were placed in independent agricultural landscapes. We compared next-generation (Illumina), i.e., short-read, and thirdgeneration (Nanopore), i.e., MinION, sequencing techniques. MinION data were preprocessed using traditional and Nanopore specific tools for comparative analysis and were evaluated in comparison to short-read sequence data with conventional processing. Based on the results, the dietary diary of bumble bee in the studied landscapes can be identified. It is known that short reads generated by next-generation sequencers have the advantage of higher quality scores while Nanopore yields longer read lengths. We show that assignments to taxonomic units yield comparable results when querying against an ITS2-specific sequence database. Thus, lower sequence quality is compensated by longer read lengths. However, the Nanopore technology is improving in terms of data quality, much cheaper, and suitable for portable applications. With respect to the studied agricultural landscapes we found that bumble bees require higher plant diversity than only crops to fulfill their foraging requirements.
• Journal Article

#### Flavor-Related Quality Attributes of Ripe Tomatoes Are Not Significantly Affected Under Two Common Household Conditions ﻿

Frontiers in Plant Science 2020; 11 p.1-14: Art. 472
Consumer complaints about the flavor of fresh tomato fruits (Solanum lycopersicum L.) have increased in the past few decades, and numerous studies have been done on the flavor of tomatoes and how it is influenced. However, it has not yet been taken into account how consumer handling affects the flavor when considering the complete post-harvest chain—from retailer (distributor) to retail to consumer. In this study, the impact of two household storage regimes on the volatile profile and important flavorrelated compounds were examined, considering the entire post-harvest handling. New breeding lines ($n = 2$) and their parental cultivars ($n = 3$) were evaluated. Fruits were harvested ripe and stored at 12.5°C for 1 day, at 20°C for 2 days, and afterward at either 20 or 7°C for another 4 days. The aroma volatile profile was measured using GC-MS and GC-FID. A trained panel was used to characterize the sensory attributes of the fruits. In both storage regimes, the relative amount of hexanal increased during the storage period in three of the five cultivars/breeding lines while benzaldehyde was the only volatile compound that decreased significantly in four cultivars/breeding lines. The relative concentration of the precursors of lipid-derived volatiles—linoleic (C18:2) and linolenic (C18:3) acid—did not change in both storage regimes. The lycopene and b-carotene contents increased slightly during storage (20 and 7°C), as the carotenoidderived volatile 6-methyl-5-hepten-2-one did. The fructose and glucose concentrations did not vary significantly, while the content of total soluble solids increased during both storage regimes. No significant difference could be found between the fruits stored at 20 or 7°C for 4 days after the post-harvest handling in all the parameters analyzed, including the sensory analysis, considering all cultivars/breeding lines. A storage temperature of 7°C is not detrimental for the flavor of ripe fruits under the experimental conditions used. The genetic background of the studied cultivars/breeding lines have a higher impact on the flavor variation than the two common household storage conditions when storing ripe fruits and taking the entire post-harvest handling into account.
• Journal Article

#### Nutrition Policy and Individual Struggle to Eat Healthily: The Question of Public Support ﻿

Nutrients 2020; 12(2) p.1-21: Art. 516
The evidence for the effectiveness of nutrition policy interventions is growing. For the implementation of such interventions, social acceptability is crucial. Therefore, this study provides insight into public support for nutrition policy measures such as labelling and taxation. Further it analyses the level of acceptance in a quantitative segmentation approach. A new element to our approach is the comparison of different policy instruments, focusing on the interaction between policy acceptance and the perceived individual struggle to eat healthily. The survey was conducted in November 2017 and a total of 1035 German consumers are included in the data. The results indicate that the majority of German citizens accept nutrition policy interventions. Based on a cluster analysis, five different target groups according to the general acceptance of policy interventions and their own struggle to eat healthily are derived. The five-cluster solution reveals that both consumers who tend to eat a healthy diet as well as those who have problems with their diet support nutritional interventions. This shows that the perceived own struggle to eat healthily does not predict whether consumers accept nutrition policy interventions.
• Journal Article

#### Impact of Environmental Conditions and Agronomic Practices on the Prevalence of Fusarium Species Associated with Ear- and Stalk Rot in Maize ﻿

Pathogens 2020; 9(3) p.1-17: Art. 236
Fusarium species are common pathogens on maize and reduce the product quality through contamination with mycotoxins thus jeopardizing safety of both animal feed and human food products. Monitoring of Fusarium infected maize ears and stalks was conducted in Germany to determine the range of Fusarium species present in the field and to assess the impact of tillage, crop rotation, and weather conditions on the frequency of Fusarium species. From 2016 till 2018, a total of 387 infected ears and 190 stalk segments from 58 locations in Germany were collected. For each sample location, site-specific agronomic data on tillage and previous crops as well as meteorological data on precipitation, air temperature, and relative humidity during the vegetation period were recorded. The most frequent Fusarium species detected in maize ears were Fusarium graminearum, F. verticillioides and F. temperatum, whereas, F. graminearum, F. equiseti, F. culmorum, and F. temperatum were the species prevailing on maize stalks. Differences in the local species composition were found to be primarily associated with weather variations between the years and the microclimate at the different locations. The results indicate that mean temperature and precipitation in July, during flowering, has the strongest impact on the local range of Fusarium spp. on ears, whereas the incidence of Fusarium species on stalks is mostly affected by weather conditions during September. Ploughing significantly reduced the infection with F. graminearum and F. temperatum, while crop rotation exerted only minor effects.
• Journal Article

#### Horses as a Crucial Part of One Health ﻿

Veterinary Sciences 2020; 7(1) p.1-17: Art. 28
One Health (OH) is a crucial concept, where the interference between humans, animals and the environment matters. This review article focusses on the role of horses in maintaining the health of humans and the environment. Horses’ impact on environmental health includes their influence on soil and the biodiversity of animal and plant species. Nevertheless, the effect of horses is not usually linear and several factors like plant–animal coevolutionary history, climate and animal density play significant roles. The long history of the relationship between horses and humans is shaped by the service of horses in wars or even in mines. Moreover, horses were essential in developing the first antidote to cure diphtheria. Nowadays, horses do have an influential role in animal assisted therapy, in supporting livelihoods in low income countries and as a leisure partner. Horses are of relevance in the spillover of zoonotic and emerging diseases from wildlife to human (e.g., Hendra Virus), and in non-communicable diseases (e.g., post-traumatic osteoarthritis in horses and back pain in horse riders). Furthermore, many risk factors—such as climate change and antimicrobial resistance—threaten the health of both horses and humans. Finally, the horse is a valuable factor in sustaining the health of humans and the environment, and must be incorporated in any roadmap to achieve OH. Keywords:
• Journal Article

#### Use of urease and nitrification inhibitors to reduce gaseous nitrogen emissions from fertilizers containing ammonium nitrate and urea ﻿

Global Ecology and Conservation 2020; 22 p.1-11: Art. e00933
Nitrogen (N) fertilizers increase agricultural yields, but also lead to the release of thegreenhouse gases nitrous oxide (N$_2$O) and ammonia (NH$_3$). This not only reduces the efficiency of N use, but also results in climate change and loss of biodiversity. The use ofnitrification inhibitors may improve the efficiency of N use and reduce the emission ofgreenhouse gases. We tested three inhibitors (NZONE MAX, Piadin and $N$-($n$-butyl) thio-phosphoric triamide (NBPT)) added to two common N fertilizers (urea and urea ammo-nium nitrate (UAN)) and determined emissions of CO$_2$, N$_2$O and NH$_3$ to evaluate theeffectiveness of these three inhibitors and to improve our understanding of the soil ni-trogen cycle. NBPT effectively reduced NH3volatilization by 50% (from 3.0 g NH$_3$-N m$^{-2}$ in urea alone to 1.4 g NH$_3$-N m$^{-2}$ in urea + NBPT). Piadin decreased N$_2$O emissions (from 0.98 g N$_2$O-N m$^{-2}$ in urea alone to 0.15 g N$_2$O-N m$^{-2}$ in urea + Piadin and from 0.81 g N$_2$O-N m$^{-2}$ in UAN alone to 0.39 g N$_2$O-N m$^{-2}$ in UAN + Piadin) by inhibiting the conversion of NH$_4^+$ to NO$_3^-$. However, although Piadin was found to be an effective nitrification inhibitor,the risk of higher NH$_3$ emissions (from 3.0 g NH$_3$-N m$^{-2}$ in urea alone to 4.5 g NH$_3$-N m$^{-2}$ in urea + Piadin) with the addition of Piadin cannot be neglected in environmental andeconomical evaluations.
• Journal Article

#### Using a Citizen Science Approach with German Horse Owners to Study the Locomotion Behaviour of Horses on Pasture ﻿

Sustainability 2020; 12(5) p.1-12: Art. 1835
Engaging farmers as citizen scientists may be a cost-effcient way to answering applied research questions aimed at more sustainable land use. We used a citizen science approach with German horse farmers with a dual goal. Firstly, we tested the practicability of this approach for answering ‘real-life’ questions in variable agricultural land-use systems. Secondly, we were interested in the knowledge it can provide about locomotion of horses on pasture and the management factors influencing this behaviour. Out of 165 volunteers, we selected 40 participants to record locomotion of two horses on pasture and provide information on their horse husbandry and pasture management. We obtained complete records for three recording days per horse from 28 participants, resulting in a dataset on more individual horses than any other Global Positioning System study published in the last 30 years. Time spent walking was greatest for horses kept in box-stall stables, and walking distance decreased with increasing grazing time. This suggests that restrictions in pasture access may increase stress on grass swards through running and trampling, severely challenging sustainable pasture management. Our study, involving simple technology, clear instructions and rigorous quality assessment, demonstrates the potential of citizen science actively involving land managers in agricultural research.
• Journal Article

#### Agroforestry Management Systems Drive the Composition, Diversity, and Function of Fungal and Bacterial Endophyte Communities in Theobroma Cacao Leaves ﻿

Microorganisms 2020; 8(3) p.1-24: Art. 405
Cacao (Theobroma cacao L.) is one of the most economically important crops worldwide. Despite the important role of endophytes for plant growth and health, very little is known about the effect of agroforestry management systems on the endophyte communities of T. cacao. To close this knowledge gap, we investigated the diversity, community composition, and function of bacterial and fungal endophytes in the leaves of T. cacao trees growing in five major cacao-growing regions in the central region of Cameroon using DNA metabarcoding. Fungal but not bacterial alpha diversity measures differed significantly between the agroforestry management systems. Interestingly, less managed home-garden cacao forests harbored the lowest fungal richness and diversity. Our results suggest that the composition of bacterial and fungal endophyte communities is predominantly affected by agroforestry management systems and, to a lesser extent, by environmental properties. The core microbiome detected comprised important fungal phytopathogens, such as Lasiodiplodia species. Several predicted pathways of bacterial endophytes and functional guilds of fungal endophytes differed between the agroforest systems which might be attributed to bacteria and fungi specifically associated with a single agroforest. Our results provide the basis for future studies on foliar fungal and bacterial endophytes of T. cacao and their responsiveness towards agroforestry management systems.
• Journal Article

#### Plant–pollinator interactions along an urbanization gradient from cities and villages to farmland landscapes ﻿

Ecosphere 2020; 11(2) p.1-12: Art. e03020
Urbanization affects pollinator diversity and plant–pollinator networks by changing resource availability locally and in the surrounding landscape. We experimentally established (N = 12) standardized plant communities in farmland, villages, and cities to identify the relative role of local and landscape effects on plant–pollinator communities along this urbanization gradient. We found that the number of flower visits by solitary bees, but not bumblebees, was highest in cities and lowest in farmland, with villages being intermediate, whereas syrphid flies exhibited lowest numbers in cities. Villages supported the richest pollinator communities, as they appeared to benefit from both farmland and city communities. Plant–pollinator network metrics such as robustness, interaction evenness, and interaction diversity decreased with increasing urbanization, although local plant richness increased toward urban areas. In conclusion, pollinator communities were most diverse and stable in farmland and village sites, despite the high plant richness in cities. The different composition of pollinator communities along the urbanization gradient suggests considering all three landscape types for conservation schemes.