Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana

Plant‐based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C‐terminal fused to the heavy chain of 14D9 (vac‐Abs) and compared with secreted and ER‐retained variants (sec‐Ab, ER‐Ab, respectively). Accumulation of ER‐ and vac‐Abs was 10‐ to 15‐fold higher than sec‐Ab. N‐glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec‐Ab while vac‐Abs carried mainly oligomannosidic (Man 7‐9) next to GnGnXF forms. Paucimannosidic glycans (commonly assigned as typical vacuolar) were not detected. Confocal microscopy analysis using RFP fusions showed that sec‐Ab‐RFP localized in the apoplast while vac‐Abs‐RFP were exclusively detected in the central vacuole. The data suggest that vac‐Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N‐glycans). Importantly, vac‐Abs were correctly assembled and functionally active. Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post‐translational modifications, but also point to a reconsideration of current concepts in plant glycan processing.     

Plant‐based oral vaccines against zoonotic and non‐zoonotic diseases

The shared diseases between animals and humans are known as zoonotic diseases and spread infectious diseases among humans. Zoonotic diseases are not only a major burden to livestock industry but also threaten humans accounting for >60% cases of human illness. About 75% of emerging infectious diseases in humans have been reported to originate from zoonotic pathogens. Because antibiotics are frequently used to protect livestock from bacterial diseases, the development of antibiotic‐resistant strains of epidemic and zoonotic pathogens is now a major concern. Live attenuated and killed vaccines are the only option to control these infectious diseases and this approach has been used since 1890. However, major problems with this approach include high cost and injectable vaccines is impractical for >20 billion poultry animals or fish in aquaculture. Plants offer an attractive and affordable platform for vaccines against animal diseases because of their low cost, and they are free of attenuated pathogens and cold chain requirement. Therefore, several plant‐based vaccines against human and animals diseases have been developed recently that undergo clinical and regulatory approval. Plant‐based vaccines serve as ideal booster vaccines that could eliminate multiple boosters of attenuated bacteria or viruses, but requirement of injectable priming with adjuvant is a current limitation. So, new approaches like oral vaccines are needed to overcome this challenge. In this review, we discuss the progress made in plant‐based vaccines against zoonotic or other animal diseases and future challenges in advancing this field.     

Community participation and ecological criteria for selecting species and restoring natural capital with native species in the Sahel

Combining community needs and preferences with dryland plant expertise in order to select suitable native species for large‐scale natural capital restoration is the approach that has been successful in the Sahel as part of Africa's Great Green Wall program. In order to increase plant diversity and restore degraded land, we investigated four cross‐border regions of Mali, Burkina Faso, and Niger, all located in dryland ecosystems of the Sahel. In 120 beneficiary village communities, with a total population of over 50,000 farmers, including 51% women, participatory diagnostic meetings were conducted, leading to the selection of 193 plant species, most of which were mainly used for food, medicine, fodder, and fuel. Of these, 170 were native and considered suitable for enriching and restoring those village lands. The most environmentally well‐adapted and economically relevant species were prioritized, quality seeds were collected, and nursery seedlings produced under technical supervision of villages. From 2013 to 2015, 55 woody and herbaceous species were planted to initiate restoration of 2,235 ha of degraded land. On average, 60% of seedlings survived and grew well in the field after three rainy seasons. Due to its multiple uses, including gum arabic production, Acacia senegal was preferred by local people in most cases, accounting for 30% of seedlings planted. Such promising results, in an effort to restore degraded land for and with the help of thousands of farmers, could not have been achieved without the combination of scientific plant expertise and efficient rural capacity development, underpinned by high levels of community engagement.     

The role of fish-poultry integration on fish growth performance,yields and economic benefits among smallholder farmers in sub-Saharan Africa,Tanzania

Aquaculture practices from sub-Saharan Africa are characterised by low production, owing to improper technology. Production can be increased through integrating fish farming with other existing on-farm activities, particularly livestock husbandry. We assessed the role of fish-poultry integration on all male Nile tilapia, Oreochromis niloticus growth performance, yields and economic benefits among smallholder farmers in sub-Saharan Africa, Tanzania. The study also compared phytoplankton species composition, abundance and biomass between the fish-poultry integration and non-integrated system. After 180 days of the experiment, all male O. niloticus cultured under fish-poultry integration exhibited significantly higher growth rates than those in the non-integrated system (p < 0.05). Gross fish yield (GFY), net fish yield (NFY) and net annual yields (NAY) obtained from fish-poultry integration were significantly higher than those from non-integrated system (p < 0.05). Partial enterprise budget analysis revealed that fish-poultry integration was more profitable than the non-integrated system. Moreover, fish-poultry integrated system produced significantly higher phytoplankton abundance and biomass than those from the non-integrated system. Results demonstrate that rural smallholder farmers can achieve higher growth rate, farm net yields and income by integrating all male O. niloticus with other on-farm activities than practising a stand-alone fish culture system.     

Health status of the lizard Podarcis siculus (Rafinesque-Schmaltz, 1810) subject to different anthropogenic pressures

The terrestrial lizard Podarcis siculus is the most abundant reptile in Italy, where is considered a ubiquitous species. This lizard is widely distributed from the islands to the Apennines, from cultivated fields to anthropized areas such as gardens and city parks. For this reason, these animals are exposed to extensive physical and chemical stresses, as well as to the possibility of coming into contact with industrial pollutants and substances used in agricultural practices. Here, we review the health status of lizard specimens inhabiting natural non-anthropized areas and fields devoted to organic farming, considering the condition of (1) liver, representing the main detoxifying organ, directly influenced by feeding, and (2) gonads, essential for reproduction and, therefore, for the survival of the species. The morphological and biomolecular condition of these organs was then compared with those obtained from lizard specimens experimentally treated with nonylphenol, a co-formulant of many insecticides and plant protection products widely used in conventional farming, known to have harmful estrogenic effects. Taken together, data demonstrate that lizards inhabiting manured soil are in good health status and show a regular morphology of liver, testis, and ovary. Animals are found to be less exposed to the toxic heavy metals cadmium and lead if compared with specimens collected in areas not devoted to agriculture, but probably more exposed to vehicular traffic. However, manure, as well as nonylphenol, exerts a xeno-estrogenic effect, particularly evident in male specimens, more sensitive to estrogenic contamination.     

Water footprint assessment of sheep farming systems based on farm survey data

Water scarcity is among the main challenges making vulnerable the livestock farming systems in drylands. The water footprint (WF) indicator was proposed as a metric to measure the impacts of livestock production on freshwater resources. Therefore, this study aimed to assess water use in five different Tunisian sheep production systems using the Water Footprint Network methodology. The primary data were obtained from 1050 sheep farms located in 13 Tunisian provinces. A multivariate analysis was performed to characterize the different farming systems. A validation step of the WF modeled values of sheep meat was conducted in 12 sheep farms belonging to two different farming systems. This was done through year-round monitoring of on-farm practices using water metres and recording equipment’s taking into account the direct and indirect water use. The typology analysis came up with five sheep farming systems that are the mixed sheep-cereal (MSC), the agro-sylvo-pastoral (ASP), the agro-pastoral (AP), the extensive agro-pastoral (EAP) and the mixed sheep-olive tree farming systems. The WF of sheep meat produced under the target farming systems ranged from 8654 to 13 056 l/kg live weight. The evaluation of WF of five different sheep production systems figured out that sheep raised under the EAP farming system had the greatest WF per ton of live animal. However, the ASP farming system exhibited the lowest WF. Water used to grow feedstuffs for sheep production accounts for 98% of the total WF of sheep. The green WF accounts for more than 92% of the total WF in all farming systems. Results of monitoring water use at farm scale show that the modeled values of WF are overestimated by an average of 23.3% and 24.1% for the selected farms assigned to the MSC and AP farming systems, respectively. Water use for sheep production is high in most of the Tunisian farms. Therefore, the general assumption that ‘meat production is a driver of water scarcity’ is supported and should be considered as an important focal point in agricultural and water policies. Particular attention should be given to forage crops with low WFs and high contribution to dry matter to provide ration with low WF. The efficient use of green water along the meat value chain is essential to minimize the depletion of blue water resources and to reduce the economic dependency on virtual water through the import of feedstuffs.     

Direct seeding and outplantings in drylands of Argentinean Patagonia: estimated costs,and prospects for large‐scale restoration and rehabilitation

In large areas of the world that are deeply scarred by desertification and hampered by low capacity for natural regeneration, the scaling up of ecological restoration and rehabilitation can be achieved only if it is low in cost with high return on investment, and shows promise of providing long‐lasting social‐economic as well as ecological benefits. In the Monte Austral region of Patagonia Argentina, concerted efforts are underway to facilitate scaling up of ecological restoration and rehabilitation practices. Here, we evaluate financial costs and preliminary results of direct seeding as compared to outplanting of nursery‐grown seedlings of three native species (Atriplex lampa, Senecio subulatus var. subulatus, and Hyalis argentea var. latisquama) considered to be high‐priority dryland framework species. Comparative success is expressed in terms of plant survival and in monetary terms. The three candidate species showed low survival rates, ranging from 4.3 to 22.3%, after the first summer following direct seeding. In contrast, survival rates for planted seedlings of the same three taxa varied between 84 and 91%, after the first summer following reintroduction. However, cost of direct seeding varied between 1,693 and 1,772 US$ less per hectare, that is, 64% less than the cost of outplanting nursery seedlings. Therefore, in the search for ways to scale up ecological restoration and rehabilitation in drylands, direct seeding should receive more attention. We discuss the social and ecological perspectives and the way forward for direct seeding techniques in Patagonia. We also consider how costs could be reduced and effectiveness improved in large‐scale efforts.     

Influence of fire and mechanical sagebrush reduction treatments on restoration seedings in Utah,United States

Overabundance of woody plants in semiarid ecosystems can degrade understory herbaceous vegetation and often requires shrub reduction and seeding to recover ecosystem services. We used meta‐analysis techniques to assess the effects of fire and mechanical shrub reduction over two post‐treatment timeframes (1–4 and 5–10 years) on changes in cover and frequency of 15 seeded species at 63 restoration sites with high potential for recovery. Compared to mechanical treatments, fire resulted in greater increases in seeded species. Native shrubs did not increase, and forbs generally declined over time; however, large increases in perennial grasses were observed, suggesting that seeding efforts contributed to enhanced understory herbaceous conditions. We found greater increases in a few non‐native species than native species across all treatments, suggesting the possibility that interference among seeded species may have influenced results of this regional assessment. Differences among treatments and species were likely driven by seedbed conditions, which should be carefully considered in restoration planning. Site characteristics also dictated seeded species responses: while forbs showed greater increases in cover over the long term at higher elevation sites considered to be more resilient to disturbance, surprisingly, shrubs and grasses had greater increases in cover and frequency at lower elevation sites where resilience is typically much lower. Further research is needed to understand the causes of forb mortality over time, and to decipher how greater increases of non‐native relative to native seeded species will influence species diversity and successional trajectories of restoration sites.     

Lao peasants on the move: Pathways of agrarian change in Laos

Grant Evans’ body of work on Laos consistently sought to understand how peasant society responded to political, social and economic change. As the Lao peasantry became ever more integrated both nationally and within the wider Southeast Asian region, and relations of market exchange came to dominate rural life, Evans maintained the need to reflect on Laos’ peasant roots to understand resulting pathways of agrarian change. This paper reconnects with this discussion in the contemporary Lao context, focusing on three dimensions of mobility: the widening spatial and sectoral ‘footprints’ of rural livelihoods; physical movement within and beyond rural settings in Laos; and reflections on issues of differentiation. We first track the political and policy changes that have most strongly influenced rural life since Evans’ early research on collectivisation, before exploring ways in which spatial integration, subsistence‐to‐commercial transition and migration have situated the remnants of Lao peasant society in a world of flows.