Farming practices and biodiversity: Evidence from a Mediterranean semi-extensive system on the island of Lemnos (North Aegean, Greece).

The management of agroecosystems affects biodiversity at all levels from genetic to food-web complexity. Low-input farming systems support higher levels of genetic, species and habitat diversity than high-input, industrial ones. In Greece, as in other Mediterranean countries, the role of traditional farming practices has been underlined in studies concerning conservation in agricultural landscapes. With this study, we aim to provide evidence for the potential of semi-extensive farming for biodiversity conservation at landscape-scale, focusing on Lemnos, a medium-sized island in the North Aegean. Evidence was gathered by species- and community-level local-scale surveys on various trophic levels (vascular plants, arthropods, birds). The surveys took place in 2018 and 2019 in 25 sampling areas comprising 106 plots of 100 m2 (vascular plants, arthropods) and 57 points where bird species were recorded. The plots were classified into three landscape types: mosaic agriculture, mixed rangelands and uniform rangelands. The relevés of Lemnos farmlands were assigned to plant communities of 18 phytosociological alliances, grouped into 12 classes. The most abundant arthropods were Coleoptera, Chilopoda, and Hymenoptera, followed by Opiliones and Isopoda, while 133 different bird species were recorded in total, including the recording for the first time on the island of five species. Farming on Lemnos is rather extensive compared to most agricultural landscapes of Europe. Our approach has demonstrated that, given the geographic characteristics of the area, the measured data reveal very high biodiversity. Our explorative findings suggest that moderate seasonal grazing, the mixed habitat mosaic with ecotones, fallow and stubble fields at the landscape scale, and the small size of fields, the kinds of crop, and farm-scale crop diversification, like mixed cultivation and crop rotation, are key practices supporting this diversity. These explorative findings are considered as a first step providing the baseline for future assessments. A wider effort, for systematic evaluation of the impacts of farming practices to biodiversity, is required, as part of a subsidized agri-environmental scheme and/or through a market-oriented product certification system for the area.

Genetic differentiation of mainland-island sheep of Greece: Implications for identifying candidate genes for long-term local adaptation.

In Greece, a number of local sheep breeds are raised in a wide range of ecological niches across the country. These breeds can be used for the identification of genetic variants that contribute to local adaptation. To this end, 50k genotypes of 90 local sheep from mainland Greece (Epirus, n = 35 and Peloponnesus, n = 55) were used, as well as 147 genotypes of sheep from insular Greece (Skyros, n = 21), Lemnos, n = 36 and Lesvos, n = 90). Principal components and phylogenetic analysis along with admixture and spatial point patterns analyses suggested genetic differentiation of 'mainland-island' populations. Genome scans for signatures of selection and genome-wide association analysis (GWAS) pointed to one highly differentiating marker on OAR4 (FST = 0.39, FLK = 21.93, FDR p-value = 0.10) that also displayed genome wide significance (FDR p-value = 0.002) during GWAS. A total number of 6 positional candidate genes (LOC106990429, ZNF804B, TEX47, STEAP4, SRI and ADAM22) were identified within 500 kb flanking regions around the significant marker. In addition, two QTLs related to fat tail deposition are reported in genomic regions 800 kb downstream the significant marker. Based on gene ontology analysis and literature evidence, the identified candidate genes possess biological functions relevant to local adaptation that worth further investigation.

Clustering patterns mirror the geographical distribution and genetic history of Lemnos and Lesvos sheep populations.

Elucidating the genetic variation and structure of Lemnos and Lesvos sheep is critical for maintaining local genetic diversity, ecosystem integrity and resilience of local food production of the two North Aegean islands. In the present study, we explored genetic diversity and differentiation as well as population structure of the Lemnos and Lesvos sheep. Furthermore, we sought to identify a small panel of markers with the highest discriminatory power to assign animals across islands. A total number of n = 424 (n = 307, Lemnos and n = 117, Lesvos) ewes, sampled from n = 24 herds dispersed at different geographic regions on the two islands, were genotyped with the 50K SNP array. Mean observed heterozygosity was higher (but not statistically significantly different) in Lesvos than in Lemnos population (0.384 vs. 0.377) while inbreeding levels were higher in Lemnos than Lesvos herds (0.065 vs. 0.031). Results of principal components along with that of admixture analysis and estimated genetic distances revealed genetic clusters corresponding to Lesvos and Lemnos origin and the existence of infrastructure within islands that were associated with geographical isolation and genetic history of the studied populations. In particular, genetic analyses highlighted three geographically isolated herds in Lemnos that are located at mountainous areas of the island and are characterized as representatives of the local sheep by historic data and reports. Admixture analysis also showed a shared genetic background between Lemnos and Lesvos sheep attributable to past gene flow. Little overall genetic differentiation was detected between the two island sheep populations, while 150 discriminatory SNPs could accurately assign animals to their origin. Present results are comparable with those reported in the worldwide sheep breeds, suggesting geography related genetic patterns across and within islands and the existence of the local Lemnos sheep.

Feeding of carob (Ceratonia siliqua) to sheep infected with gastrointestinal nematodes reduces faecal egg counts and worm fecundity.

The present study explored the anthelmintic effects of condensed tannins (CT) in carob (Ceratonia siliqua) pods fed to sheep against gastrointestinal nematodes. Three independent in vivo trials tested whether i) carob pod (CaBP)-containing feed had an anthelmintic effect and if yes, which was the optimal concentration in the diet; ii) whether this effect could be attributed to tannins through the polyethylene glycol (PEG) test and iii) whether there were any synergistic effects when combined with another tannin-containing feed (e.g. sainfoin). In all trials 6-month old nematode-naive lambs, experimentally infected with both Haemonchus contortus and Trichostrongylus colubriformis, were used. Faecal egg counts (FEC) were performed regularly and at the end of each trial adult worm counts (AWC) and female worm fecundity were recorded. In trial 1, 35 lambs (five groups of seven lambs) were fed different CaBP concentrations ranging from 0% to 12 % w/w. FEC declined up to 39.2 % only in the group fed with 12 %CaBP, while a declining trend (P < 0.06) was demonstrated for the AWC of T. colubriformis, which was associated with the increasing concentration of CaBP in feed. Female worm fecundity was reduced in groups fed CaBP for both parasites, however this was only significant for H. contortus (P < 0.001), in a dose dependent manner. In trial 2, four groups of six infected lambs each were used, which received the carob diets CaBP or CaBP + PEG, and the tannin-free diets with or without PEG (C or C + PEG). Results showed that FEC of Groups C, C + PEG, and CaBP + PEG were comparable throughout the trial, while the group receiving only CaBP showed lower FEC from DAY 25 onwards. AWC showed a reduction (67.7 %) only for H. contortus (P < 0.03). Reversal of the anthelmintic effect of CaBP after PEG administration suggested that CT contributed to the anthelmintic action. However, no effect of CaBP was observed on T. colubriformis AWC and on female worm fecundity for both species. Finally, for trial 3 four groups of six lambs each received a diet based on CaBP, sainfoin (S) or a combination (CaBP + S) and were compared to a control (C) diet of lucerne. On DAY 37 FEC values in groups CaBP + S and S tended to be lower compared to the two other groups (C, CaBP), while for AWCs no significant differences were observed for both parasites. The fecundity of H. contortus and T. colubriformis demonstrated significant differences between the treated and control groups, with lower values in the animals receiving CaBP + S. Overall, the results supported the hypothesis that carob had an anthelmintic effect due to its CT, but there was no clear indication of a synergistic effect with sainfoin.

Greek Graviera Cheese Assessment through Elemental Metabolomics-Implications for Authentication, Safety and Nutrition.

This study presents the comprehensive elemental profile of Greek Graviera (Gruyère) cheeses. In total, 105 samples from nine different geographic regions produced from sheep, goat and cow milk and their mixtures were assessed. Elemental signatures of 61 elements were investigated for determination of geographic origin and milk type. Regional and milk type classification through Linear Discriminant Analysis was successful for almost all cases, while a less optimistic cross validation exercise presented lower classification rates. That points to further research using a much larger sample set, increasing confidence for cheese authentication utilizing also bioinformatics tools under development. This is the first study reporting signatures of 61 elements in dairy products including all sixteen rare earth elements and all seven precious metals. Safety and quality were assessed regarding toxic and nutritive elements. According to both EU and USA regulations and directives, Graviera is a nutritional source for trace and macro elements with low levels of toxic elements.

Challenges for local breed management in Mediterranean dairy sheep farming: insights from Central Greece.

Local breeds are recognized as an important element for the maintenance of various and specific farming systems. Challenges for local breeds' management, in a context of crossbreeding with exotic highly productive breeds, have been mainly studied in tropical countries. However, similar situation and challenges are likely to exist in Mediterranean countries subjected to climatic and feed scarcity issues. The objective of this work is to identify the challenges for local breed management in a regional context of informal crossbreeding with highly productive breeds. For this purpose, the case of dairy sheep farming in the region of Thessaly, in Central Greece, was examined. Semi-structured interviews were performed in 46 farms and processed through hierarchical classification on principal components. A follow-up on seven farms raising the Karagouniko sheep breed, the main local breed of the region, was carried on during one milk campaign. Results showed that a diversity of breeding strategies involving local purebred and crossbred flocks coexist in the region. The Karagouniko breed is facing several challenges. The supply in exotic breeding males and their crosses could be wide-scaled and involved a diversity of operators, whereas the supply in breeding males of Karagouniko breed was restrained to between-farm supply among flocks under milk recording scheme. In addition, the heterogeneity of access and quality of collective rangelands affected the farming of Karagouniko breed ewes, whose purebred flocks were significantly associated with the grazing on native grasslands. Finally, unfavorable dairies' policies led Karagouniko farmers to seek higher flock milk production through levers that could impact the vulnerability of the farm, such as earlier lambing period or earlier weaning age. Farmers also questioned the use of highly productive breeds as a potential lever to reach higher flock milk production.

Differences in sheep and goats milk fatty acid profile between conventional and organic farming systems.

The objective of this study was to investigate whether there is a difference in chemical composition and particularly in fatty acid (FA) profile, with emphasis on cis-9, trans-11 CLA, of milk obtained from conventional and organic dairy sheep and goats farms under the farming conditions practiced in Greece. Four dairy sheep and four dairy goat farms, representing common conventional production systems and another four dairy sheep and four dairy goat farms, organically certified, representing organic production and feeding systems were selected from all over Greece. One hundred and sixty two individual milk samples were collected from those farms in January-February 2009, about three months after parturition. The milk samples were analyzed for their main chemical constituents and their FA profile. The results showed that the production system affected milk chemical composition: in particular fat content was lower in the organic sheep and goats milk compared with the corresponding conventional. Milk from organic sheep had higher content in MUFA, PUFA, alpha-LNA, cis-9, trans-11 CLA, and omega-3 FA, whereas in milk from organic goats alpha-LNA and omega-3 FA content was higher than that in conventional one. These differences are, mainly, attributed to different feeding practices used by the two production systems. The results of this study show that the organic milk produced under the farming conditions practiced in Greece has higher nutritional value, due to its FA profile, compared with the respective conventional milk.