Spatial, temporal and network analyses provide insights into the dynamics of the bacterial communities associated with two species of Caribbean octocorals and indicate possible key taxa
Despite the current decline of scleractinian coral populations, octocorals are thriving on reefs in the Caribbean Sea and western North Atlantic Ocean. These cnidarians are holobiont entities, interacting with a diverse array of microorganisms. Few studies have investigated the spatial and temporal stability of the bacterial communities associated with octocoral species and information regarding the co-occurrence and potential interactions between specific members of these bacterial communities remain sparse. To address this knowledge gap, this study investigated the stability of the bacterial assemblages associated with two common Caribbean octocoral species, and across time and geographical locations and performed network analyses to investigate potential bacterial interactions. Results demonstrated that general inferences regarding the spatial and temporal stability of octocoral-associated bacterial communities should not be made, as host-specific characteristics may influence these factors. In addition, network analyses revealed differences in the complexity of the interactions between bacteria among the octocoral species analyzed, while highlighting the presence of genera known to produce bioactive secondary metabolites in both octocorals that may play fundamental roles in structuring the octocoral-associated bacteriome.
Secondary metabolites from endophytic fungi: Production, methods of analysis, and diverse pharmaceutical potential
The synthesis of secondary metabolites is a constantly functioning metabolic pathway in all living systems. Secondary metabolites can be broken down into numerous classes, including alkaloids, coumarins, flavonoids, lignans, saponins, terpenes, quinones, xanthones, and others. However, animals lack the routes of synthesis of these compounds, while plants, fungi, and bacteria all synthesize them. The primary function of bioactive metabolites (BM) synthesized from endophytic fungi (EF) is to make the host plants resistant to pathogens. EF is a group of fungal communities that colonize host tissues' intracellular or intercellular spaces. EF serves as a storehouse of the above-mentioned bioactive metabolites, providing beneficial effects to their hosts. BM of EF could be promising candidates for anti-cancer, anti-malarial, anti-tuberculosis, antiviral, anti-inflammatory, etc. because EF is regarded as an unexploited and untapped source of novel BM for effective drug candidates. Due to the emergence of drug resistance, there is an urgent need to search for new bioactive compounds that combat resistance. This article summarizes the production of BM from EF, high throughput methods for analysis, and their pharmaceutical application. The emphasis is on the diversity of metabolic products from EF, yield, method of purification/characterization, and various functions/activities of EF. Discussed information led to the development of new drugs and food additives that were more effective in the treatment of disease. This review shed light on the pharmacological potential of the fungal bioactive metabolites and emphasizes to exploit them in the future for therapeutic purposes.
Wild herbivorous mammals (genus ) host a diverse but transient assemblage of fungi
Fungi are often overlooked in microbiome research and, as a result, little is known about the mammalian mycobiome. Although frequently detected in vertebrate guts and known to contribute to digestion in some herbivores, whether these eukaryotes are a persistent part of the mammalian gut microbiome remains contentious. To address this question, we sampled fungi from wild woodrats ( spp.) collected from 25 populations across the southwestern United States. For each animal, we collected a fecal sample in the wild, and then re-sampled the same individual after a month in captivity on a controlled diet. We characterized and quantified fungi using three techniques: ITS metabarcoding, shotgun metagenomics and qPCR. Wild individuals contained diverse fungal assemblages dominated by plant pathogens, widespread molds, and coprophilous taxa primarily in Ascomycota and Mucoromycota. Fungal abundance, diversity and composition differed between individuals, and was primarily influenced by animal geographic origin. Fungal abundance and diversity significantly declined in captivity, indicating that most fungi in wild hosts came from diet and environmental exposure. While this suggests that these mammals lack a persistent gut mycobiome, natural fungal exposure may still impact fungal dispersal and animal health.
A peptidoglycan-recognition protein orchestrates the first steps of symbiont recruitment in the squid-vibrio symbiosis
In symbioses established through horizontal transmission, evolution has selected for mechanisms that promote the recruitment of symbionts from the environment. Using the binary association between the Hawaiian bobtail squid, , and its symbiont, , we explored the first step of symbiont enrichment around sites where cells will enter host tissues. Earlier studies of the system had shown that, within minutes of hatching in natural seawater, ciliated epithelia of the nascent symbiotic tissue secrete a layer of mucus in response to exposure to the cell-wall biomolecule peptidoglycan (PGN) from non-specific bacterioplankton. We hypothesized that a peptidoglycan recognition protein, EsPGRP4, is the receptor that mediates host mucus secretion by sensing the environmental PGN; earlier studies of this protein family had shown that this is the only member predicted to behave as a membrane receptor. Immunocytochemistry localized EsPGRP4 to the superficial ciliated fields of the juvenile organ. We found that production of EsPGRP4 increased over the first 48 h after hatching if the light organ remained uncolonized. When colonized by , the levels of the protein in light-organ tissue remained similar to that of hatchling organs. Pharmacologically curing the initially colonized light organ with antibiotics resulted in return of EsPGRP4 production to levels similar to light organs that had remained uncolonized since hatching. Furthermore, we found that preincubation of the tissues with an EsPGRP4 antibody decreased light organ mucus production and colonization. These findings provide evidence of an innate mechanism that underlies a crucial first step in the horizontal recruitment of bacterial symbionts.
The beauty and the yeast: can the microalgae form a borderline lichen with ?
Lichenized fungi usually develop complex, stratified morphologies through an intricately balanced living together with their algal partners, but several species are known to form only more or less loose associations with algae. These borderline lichens are still little explored although they could inform us about early stages of lichen evolution. We studied the association of the extremely halotolerant fungus with the alga , discovered in a cave in the Atacama Desert (Chile), and with . , common inhabitant of saltern brines. . forms small colonies, in which cells of . can be frequently observed, while such interaction has not been observed with . . As symbiotic interactions between and have not been reported, we performed a series of co-cultivation experiments to inspect whether these species could interact and develop more distinct lichen-like symbiotic structures. We set up co-cultures between axenic strains of (isolated both from Mediterranean salterns and from the Atacama cave) and isolates of . (from the Atacama cave) and . (isolated from Mediterranean salterns). Although we used different growth media and cultivation approaches, bright field and SEM microscopy analyses did not indicate any mutual effects in these experiments. We discuss the implications for fungal algal interactions along the transition from algal exploiters to lichen symbioses.
Phytohormone release by three isolated lichen mycobionts and the effects of indole-3-acetic acid on their compatible photobionts
Evidence is emerging that phytohormones represent key inter-kingdom signalling compounds supporting chemical communication between plants, fungi and bacteria. The roles of phytohormones for the lichen symbiosis are poorly understood, particularly in the process of lichenization, i.e. the key events which lead free-living microalgae and fungi to recognize each other, make physical contact and start developing a lichen thallus. Here, we studied cellular and extracellularly released phytohormones in three lichen mycobionts, , and , grown on solid medium, and the effects of indole-3-acetic acid (IAA) on their respective photobionts, , , sp. Using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) we found that mycobionts produced IAA, salicylic acid (SA) and jasmonic acid (JA). IAA represented the most abundant phytohormone produced and released by all mycobionts, whereas SA was released by and , and JA was released by only. With a half-life of 5.2 days, IAA degraded exponentially in solid BBM in dim light. When IAA was exogenously offered to the mycobionts' compatible photobionts at "physiological" concentrations (as released by their respective mycobionts and accumulated in the medium over seven days), the photobionts' water contents increased up to 4.4%. Treatment with IAA had no effects on the maximum quantum yield of photosystem II, dry mass, and the contents of photosynthetic pigments and α-tocopherol of the photobionts. The data presented may be useful for designing studies aimed at elucidating the roles of phytohormones in lichens.
and gen. nov. (Trebouxiophyceae Chlorophyta), two different green algal endosymbionts in freshwater sponges
Associations of freshwater sponges with coccoid green algae have been known for a long time. Two types of coccoid green algae, which are commonly assigned as zoochlorellae, are recognized by morphology: small coccoids (< 3 μm) without pyrenoids and larger -like algae (4-6 μm) with pyrenoids. Despite their wide distribution in some freshwater sponges, these green algae were never studied using a combined analysis of morphology and molecular phylogeny. We investigated several endosymbiotic strains isolated from different species, which were available in culture collections. Phylogenetic analyses of SSU and ITS rDNA sequences revealed that the strain SAG 211-40a is a member of the Chlorellaceae and represents a new species of the newly erected genus , . The phylogenetic position was confirmed by morphology and ITS-2 barcode. The endosymbionts without pyrenoid were identified as by morphology and phylogenetic analyses. The comparison with free-living strains revealed the recognition of two new species, and , which were confirmed by molecular signatures in V9 region of SSU rDNA and ITS-2 barcode.
Wanted: virtual or live! How lichens are becoming part of mass internet culture
While restrictions imposed by the COVID-19 pandemic have strongly limited and affected the work of scientists and communicators, the pandemic has also encouraged the development of new ways of networking and public engagement. People have had to resort to virtual events, with a subsequent proliferation of webinars, online meetings, and digital resources. In this situation we have had to find new ways of measuring the impact of these activities. Using the activities of the Italian Lichen Society (Società Lichenologica Italiana, SLI), Google Trends and colleagues' contributions, we evaluated the performances and impacts of virtual tools on lichenological literacy. We compared the relative success of virtual and in-person events and the effort required; we evaluated followers' appreciation of various categories of posts on SLI Facebook page; and designed a questionnaire to collate information on individual experiences of in-person and virtual events linked to lichens. As expected, online events generally required less effort to put together and deploy than in-person events and engaged more people, especially when recorded and made available online for a long time. Using online searches for the word "lichens" we found an association with national events, and there was a notable increase in membership of SLI over the last 10 years, demonstrating an increasing interest by people in lichens. Without excluding the positive effects of in-person experiences, we believe that online events offer a powerful tool to help increase interest in, and knowledge about lichens. This interest may help to mitigate the impact of anthropogenic activities on this sensitive component of the ecosystem and help human-lichen relationships.
Nitrate inhibition of N fixation and its effect on micronutrient accumulation in shoots of soybean ( L. Merr.), Bambara groundnut ( L. Vedc) and Kersting's groundnut ()
Although nitrate is known to inhibit nodulation and N fixation in symbiotic legumes, little is known about its effect on the uptake and accumulation of trace elements such as Fe, Zn, Mn and Cu. The aim of this study was to evaluate the effect of 5 mM NO supply, either with or without rhizobial inoculation, on nodulation, nodule functioning and micronutrient levels in the shoots of soybean ( L.Merr.), Bambara groundnut ( L. Vedc) and Kersting's groundnut ( Harm). The results showed reduction in plant growth, nodule formation and nodule dry matter by the supply of 5 mM NO to inoculated seedlings of all three species. Nitrate inhibition respectively caused 1.2, 1.4, and 1.5-fold decrease in nodule number per plant in Bambara groundnut, soybean and Kersting's bean, which resulted in 2.3, 3.3 and 4.5-fold reduction in nodule dry weight of the test species (in that order). The application of 5 mM NO to soybean plants also resulted in 2.5, 4.0 and 5.4-fold decrease in shoot accumulation of Fe, Zn and Mn, respectively, when compared to the purely symbiotic control plants. Furthermore, we observed 1.3, 1.8 and 1.3-fold decreases in the concentration of Zn, Mn and Cu in shoots of inoculated Bambara groundnut with NO supply, levels lower than those found in soybean. With Kersting's groundnut, shoot concentration of Fe, Zn and Cu were higher with the application of 5 mM NO to inoculated plants when compared to the purely symbiotic treatment, which was opposite to soybean. But pure NOfeeding of this species respectively resulted in 2.0, 1.4 and 1.3-fold decreases in Fe, Zn and Cu relative to inoculated NO-fed plants. Clearly, NO supply to landraces/genotypes of the three legume species did not only inhibit nodule formation and functioning, it also reduced shoot micronutrient levels in soybean and Bambara groundnut, but not Kersting's bean.
Grain yield of common bean ( L.) varieties is markedly increased by rhizobial inoculation and phosphorus application in Ethiopia
A field experiment was conducted to assess plant growth, symbiotic performance and grain yield of common bean in response to rhizobial incoculation and phosphorus application at Galalicha in Southern Ethiopia during the 2012 and 2013 cropping seasons under rain-fed conditions. The treatments consisted of 2 released common bean varieties (Hawassa Dume and Ibbado), 3 levels of inoculation (uninoculated, inoculated with strain HB-429 or GT-9) and 4 levels of phosphorus application (0, 10, 20 and 30 kg P ha) using a split-split plot design with four replications. Here, phosphorus levels, inoculation and common bean varieties were assigned as main, sub- and sub-sub treatments, respectively. The results revealed marked varietal differences in plant growth, grain yield and symbiotic performance. Of the two common bean varieties studied, Hawassa Dume generally showed superior performance in most measured parameters in 2013. inoculation significantly ( ≤ 0.05) increased plant growth, symbiotic performance and grain yield. Applying strain HB-429 to bean crop respectively increased plant growth, %Ndfa, amount of N-fixed and grain yield by 19, 17, 54 and 48% over uninoculated control. Similarly, the application of 20 kg P ha to bean plants respectively resulted in 36, 20, 96 and 143% increase in plant growth, %Ndfa, N-fixed and grain yield when compared to the control. These results clearly indicate that plant growth, symbiotic performance and grain yield of common bean can be significantly increased by inoculation and phosphorus fertilization in Ethiopia. inoculants are a cheaper source of nitrogen than chemical fertilizers and when combined with moderate phosphorus application can markedly increase grain yield for resource-poor farmers.
From isolation to application: a case study of arbuscular mycorrhizal fungi of the Arabian Peninsula
The vegetation in the Arabian Peninsula experiences drought, heat, soil salinity, and low fertility, mainly due to low phosphorus (P) availability. The beneficial mycorrhizal symbiosis between plants and arbuscular mycorrhizal fungi (AMF) is a key factor supporting plant growth under such environmental conditions. Therefore, AMF strains isolated from these soils might be useful as biotechnological tools for agriculture and revegetation practices in the region. Here we present a pioneering program to isolate, identify, and apply AMF isolated from rhizosphere soils of agricultural and natural habitats, namely date palm plantations and five native desert plants, respectively in the Southern Arabian Peninsula. We established taxonomically unique AMF species as single-spore cultures as part of an expanding collection of AMF strains adapted to arid ecosystems. Preliminary experiments were conducted to evaluate the abilities of these AMF strains to promote seedling growth of a main crop L. and a common plant L. (Druce) in the Arabian Peninsula. The results showed that inoculation with certain AMF species enhanced the growth of both plants, highlighting the potential of these fungi as part of sustainable land use practices in this region.
Demonstrating the role of symbionts in mediating detoxification in herbivores
Plant toxins constitute an effective defense against herbivorous animals. However, many herbivores have evolved adaptations to cope with dietary toxins through detoxification, excretion, sequestration, target site insensitivity and/or via behavioral avoidance. While these adaptations are often directly encoded in herbivore genomes, evidence is accumulating that microbial symbionts can reduce the dose of plant toxins by metabolizing or sequestering them prior to absorption by the herbivore. Here, we describe a few well-studied examples to assess such symbiont-mediated detoxification and showcase different approaches that have been used for their analyses. These include: (i) a host phenotypic route in which the symbiotic association is manipulated to reveal host fitness costs upon toxin exposure in the presence/absence of detoxifying symbionts, including function restoration after symbiont re-infection, (ii) a molecular microbiological approach that focuses on the identification and characterization of microbial genes involved in plant toxin metabolism, and (iii) an analytical chemical route that aims to characterize the conversion of the toxin to less harmful metabolites in vivo and link conversion to the activities of a detoxifying symbiont. The advantages and challenges of each approach are discussed, and it is argued that a multi-pronged strategy combining phenotypic, molecular, and chemical evidence is needed to unambiguously demonstrate microbial contributions to plant toxin reduction and the importance of these processes for host fitness. Given the interdisciplinary nature of the topic, we aim to provide a guideline to researchers interested in symbiont-mediated detoxification and hope to encourage future studies that contribute to a more comprehensive and mechanistic understanding of detoxification in herbivores and their symbionts.
Introduction to the proceedings of the 10 International Symbiosis Congress (Lyon, France)
Effect of nutritional condition on photosymbiotic consortium of cultured (Rhizaria, Foraminifera)
Several foraminifers found in warm and low-nutrient ocean surface water have photosynthetic algae as endosymbionts (photosymbiosis). To understand the trophic interactions, we studied , a spinose planktic foraminifer that has a dinoflagellate endosymbiont. We controlled two nutritional factors, feeding and inorganic nutrients in the seawater. The growth of the host and the symbionts and the photophysiological parameters were monitored under four experimental conditions. The results demonstrated that the holobionts primarily relied on phagotrophy for growth. The foraminifers grew considerably, and the chlorophyll content per foraminifer, which is an indicator of the symbiont population, increased in the fed groups, but not in the unfed groups. The nutrient-rich seawater used for some of the cultures made no difference in either the growth or photophysiology of the holobionts. These observations indicated that the symbionts mainly utilized metabolites from the hosts for photosynthesis rather than inorganic nutrients in the seawater. Additionally, we observed that the symbionts in the starved hosts maintained their photosynthetic capability for at least 12 days, and that the hosts maintained at least some symbionts until gametogenesis was achieved. This suggests that the hosts have to retain the symbionts as an energy source for reproduction. The symbionts may also play an indispensable role in the metabolic activities of the hosts including waste transport or essential compound synthesis. Overall, our results revealed a novel mode of photosymbiosis in planktic foraminifers which contrasts with that found in benthic photosymbiotic foraminifers and corals.
Symbiosis and the Anthropocene
Recent human activity has profoundly transformed Earth biomes on a scale and at rates that are unprecedented. Given the central role of symbioses in ecosystem processes, functions, and services throughout the Earth biosphere, the impacts of human-driven change on symbioses are critical to understand. Symbioses are not merely collections of organisms, but co-evolved partners that arise from the synergistic combination and action of different genetic programs. They function with varying degrees of permanence and selection as emergent units with substantial potential for combinatorial and evolutionary innovation in both structure and function. Following an articulation of operational definitions of symbiosis and related concepts and characteristics of the Anthropocene, we outline a basic typology of anthropogenic change (AC) and a conceptual framework for how AC might mechanistically impact symbioses with select case examples to highlight our perspective. We discuss surprising connections between symbiosis and the Anthropocene, suggesting ways in which new symbioses could arise due to AC, how symbioses could be agents of ecosystem change, and how symbioses, broadly defined, of humans and "farmed" organisms may have launched the Anthropocene. We conclude with reflections on the robustness of symbioses to AC and our perspective on the importance of symbioses as ecosystem keystones and the need to tackle anthropogenic challenges as wise and humble stewards embedded within the system.
Assessing host range, symbiotic effectiveness, and photosynthetic rates induced by native soybean rhizobia isolated from Mozambican and South African soils
Host range and cross-infectivity studies are important for identifying rhizobial strains with potential for use as inoculants. In this study, 10 native soybean rhizobia isolated from Mozambican and South African soils were evaluated for host range, symbiotic effectiveness and ability to induce high rates of photosynthesis leading to enhanced plant growth in cowpea ( L. Walp.), Bambara groundnut ( L. Verdc.), Kersting's groundnut ( Harm) and soybean ( L. Merr). The test isolates had different growth rates and colony sizes. Molecular analysis based on enterobacterial repetitive intergenic consensus (ERIC)-PCR revealed high genetic diversity among the test isolates. The results further showed that isolate TUTLBC2B failed to elicit nodulation in all test plants, just as TUTNSN2A and TUTDAIAP3B were also unable to nodulate cowpea, Kersting's bean and Bambara groundnut. Although the remaining strains formed ineffective nodules on cowpea and Kersting's bean, they induced effective nodules on Bambara groundnut and the two soybean genotypes. Bacterial stimulation of nodule numbers, nodule dry weights and photosynthetic rates was generally greater with isolates TUTRSRH3A, TUTM19373A, TUTMCJ7B, TUTRLR3B and TUTRJN5A. As a result, these isolates elicited significantly increased accumulation of biomass in shoots and whole plants of Bambara groundnut and the two soybean genotypes. Whole-plant symbiotic nitrogen (N) of soybean and Bambara groundnut was highest for the commercial strains CB756 and WB74, as well as for TUTRLR3B, TUTMCJ7B and TUTRSRH3A, suggesting that the three native rhizobial isolates have potential for use as inoculants.
Selecting elite groundnut ( L) genotypes for symbiotic N nutrition, water-use efficiency and pod yield at three field sites, using N and C natural abundance
About 70% of the groundnut ( L.) produced in Ghana is from the Guinea savanna. However, low soil nutrients, especially N, together with erratic rainfall distribution have often resulted in poor grain yield. The aim of this study was to evaluate plant growth, N-fixing efficiency, N contribution, water-use efficiency and pod yield of 21 elite groundnut genotypes in the Guinea savanna of Ghana, using the N natural abundance technique. The data revealed significant variations in plant growth, symbiotic N contribution, and pod yield among the 21 genotypes tested at each field site. Average N contribution by groundnut genotypes ranged from 48 to 108 kg N ha. Also, mean pod yield ranged from 0.58 to 2.1 t ha. Genotypes ICGV-IS 08837, ICG 6222, ICGV 03315 and NKATIESARI demonstrated superior plant growth, symbiotic N contribution and greater pod yield. In fact, ICGV-IS 08837 yielded almost 2.5 fold more than CHINESE which is the most widely cultivated variety in the region. Genotypes ICGV-IS 08837, ICG 6222, ICGV 03315 and ICGV 99247 are therefore recommended for development into varieties for the Guinea savanna of Ghana. Genotypes ICG (FDRS) 4, ICGV00362 and ICGV99247 exhibited increased water-use efficiency, but were low in N fixation and N contribution, and would therefore be good parental material in breeding programs aimed at enhancing water-use efficiency in high N-fixing genotypes.
World symbiosis day webinar - when living together is a win-win
The following information is intended for those who were unable to attend the first webinar of the International Symbiosis Society (ISS) on - 'When living together is a win-win' - a celebration of 'World Symbiosis Day' on 30 July 2020. The objective of the webinar was to disseminate information about the Society, to gather feedback, and to encourage the audience to join the Society. This introduction presents a summary of the webinar, highlighting the keynote presentations, the panel discussion, the journal Symbiosis, and the next ISS conference to be held in Lyon in 2022. In addition, we report on the discussions and feedback from participants that were collected through polls and other aspects of the webinar.
An assessment of plant growth and N fixation in soybean genotypes grown in uninoculated soils collected from different locations in Ethiopia
Achieving food and nutritional security is a major challenge in Ethiopia, especially with increasing human population and low crop productivity. Legumes offer an alternative choice to chemical fertilizers for increasing crop yields. The aim of this study was to assess, under glasshouse conditions, plant growth and symbiotic performance of uninoculated soybean genotypes planted in soils collected from different locations in Ethiopia. The results showed significant differences in plant growth and symbiotic performance among the soybean genotypes planted in different soils. There was a location-specific effect of soil on plant growth and symbiotic N nutrition of soybean. Whole-plant biomass was highest in soil from Amaro, followed by Boricha, Dorebafano, Pawe, and Mambuk. The δN values ranged from +0.82‰ for Pawe to +5.11‰ at Dorebafano. However, %Ndfa of soybean was greater in plants grown in Mambuk soil, followed by Pawe with the lowest %Ndfa being in Amaro soil. The amount of N-fixed followed similar pattern as %Ndfa. The significant interaction found between soil type and soybean genotype for plant DM, shoot N concentration, δN, %Ndfa, N-fixed and soil N-uptake clearly indicated the effect of soil factors. This study revealed the presence of native rhizobia in Ethiopian soils that are compatible with soybean. The N contribution of the soybean genotypes was variable, and strongly influenced by the soil factors.
The impacts of host association and perturbation on symbiont fitness
Symbiosis can benefit hosts in numerous ways, but less is known about whether interactions with hosts benefit symbionts-the smaller species in the relationship. To determine the fitness impact of host association on symbionts in likely mutualisms, we conducted a meta-analysis across 91 unique host-symbiont pairings under a range of spatial and temporal contexts. Specifically, we assess the consequences to symbiont fitness when in and out of symbiosis, as well as when the symbiosis is under suboptimal or varying environments and biological conditions (e.g., host age). We find that some intracellular symbionts associated with protists tend to have greater fitness when the symbiosis is under stressful conditions. Symbionts of plants and animals did not exhibit this trend, suggesting that symbionts of multicellular hosts are more robust to perturbations. Symbiont fitness also generally increased with host age. Lastly, we show that symbionts able to proliferate in- and outside host cells exhibit greater fitness than those found exclusively inside or outside cells. The ability to grow in multiple locations may thus help symbionts thrive. We discuss these fitness patterns in light of host-driven factors, whereby hosts exert influence over symbionts to suit their own needs.