Marine Pseudomonas: diving into the waves of blue biotechnology

Abstract

From marine to terrestrial environments, Pseudomonas spp. exhibit a remarkable ability not only to adapt but also thrive even amidst adverse conditions. This fact turns Pseudomonas spp. into one of the most prominent candidates for novel biotechnological solutions. Even though terrestrial isolates have been extensively studied, there is still an almost untapped source to be explored in marine Pseudomonas. Harnessing such strains offers an opportunity to discover novel bioactive compounds that could address current global challenges in healthcare and sustainable development. Therefore, this minireview aimed to provide an overview of the main recent discoveries regarding antimicrobials, antifouling, enzymes, pigments, and bioremediation strategies derived from marine isolates of Pseudomonas spp. Future research perspectives will also be discussed to foster forthcoming endeavors to explore the marine counterparts of such a prolific bacterial genus.

Introduction

Pseudomonas spp. are widely known for their biotechnological potential. These are remarkably diverse and adaptable bacteria, with more than 190 species described up to date (Silby et al. 2011, Peix et al. 2018). One of the first reports on the prospective use of Pseudomonas spp. dates from the first half of the 20th century, when the production of soluble exo-cellular pectinases in simple culture media was described (Oxford 1944). Since then, research on the biotechnological capabilities of this genus has progressively increased. This is largely due to the physiologic versatility and genetic plasticity of Pseudomonas species, which can be found in a wide range of habits. From hot desert soil to freezing Antarctic lakes, they can endure almost any kind of environmental stressors, turning them into prominent candidates for the discovery of novel bioactive compounds (Cagide et al. 2023, Zouagui et al. 2023).

The use of microbial-based alternatives to conventional processes has been gaining popularity, especially because they tend to be cost-effective and sustainable (Vuong et al. 2022). Consequently, considering the new demands that emerge as the world population exponentially increases, the role of microorganisms is anticipated in addressing issues related to food safety, bioremediation, development of new antimicrobials, new biocatalysts, and more. Within this framework, the marine environment has been proposed as a propitious source of biotechnological novelty, hence the rise of the term “Blue Biotechnology.” The intricate interplay of physical and chemical gradients shapes the evolution and adaptation of the marine microbiome, making it a valuable source of biodiversity (Rotter et al. 2021) (Fig. 1). Indeed, from 2021 to 2030, the United Nations (UN) has proclaimed the Decade of Ocean Science for Sustainable Development, which holds the premise that oceans are a crucial tool in ensuring an equitable and sustainable planet (UN 2020).

In that respect, previous studies have provided an overview of the main bioactive compounds synthesized by Pseudomonas spp. (Isnansetyo and Kamei 2009, Silby et al. 2011). Here, however, we offer an updated review centered exclusively on the biotechnology potential of marine Pseudomonas, focusing on studies published from 2009 onward. The five key research areas covered in this review (Fig. 2) include antimicrobial potential, antifouling activity, pigment production, enzyme synthesis, and bioremediation strategies. These areas illustrate the broad applications of marine biotechnology featuring Pseudomonas spp. Over the years, these areas have garnered increasing attention, particularly within the genus Pseudomonas, underscoring the relevance of studying marine isolates. This holds great promise for the discovery of novel biomolecules with potentially unique and valuable properties.

Antimicrobial activity

With the growing concern of antimicrobial-resistant pathogens, identifying new antimicrobials is an urgent research priority. Herein, marine Pseudomonas are recognized as fruitful sources of such compounds (Isnansetyo and Kamei 2009). Ever since the first report of a Pseudomonas strain isolated from tropical waters producing 2,3,4-tribromo-5(1′ hydroxy,2′,4′-dibromophenyl) pyrrole, numerous studies have subsequently highlighted the vast therapeutic potential associated with this bacterial group (Burkholder et al. 1966, Isnansetyo and Kamei 2009, Bollinger et al. 2020a).

For instance, the strain Pseudomonas aeruginosa MMG-28, isolated from marine sediments, has demonstrated antimicrobial activity toward different pathogens (Table 1). The antimicrobial was of low molecular weight with stable free radical activity that may lead to bacterial lysis (Charyulu et al. 2009). Another study focused on P. aeruginosa, this time isolated from hot spring discharge, also suggested its antimicrobial activity. The strain inhibited the growth of Mycobacterium smegmatis, and its crude extract demonstrated stability toward proteolytic enzymes and heat treatment (Zahir et al. 2014).

The strain Pseudomonas putida W15Oct28, isolated from a stream, was active against clinically relevant bacteria and fungi. The compound was highly hydrophobic, but its structure was not disclosed. The strain’s draft genome revealed the presence of type IV and VI secretion systems and several other gene clusters that encode secondary metabolites that were likely involved with antimicrobial activity (Ye et al. 2014).

The strain Pseudomonas sp. UJ-6, isolated from seawater, demonstrated antibacterial activity toward methicillin-resistant Staphylococcus aureus (MRSA). The antimicrobial compound was identified as 1-acetyl-β-carboline, and the minimum inhibitory concentration (MIC) ranged from 160 to 640 μg/ml against MRSA. This compound was also active against methicillin-sensitive S. aureus (MSSA) with MICs ranging from 16 to 128 μg/ml (Lee et al. 2013). Further, the cell-free supernatant (CFS) of Pseudomonas sp. CMF-2, isolated from jellyfish, was active against S. aureus, Escherichia coli, Bacillus subtilis, and Candida albicans. The protein CAP-1 was isolated and remained stable in a wide range of temperature (20°C–80°C) and pH (2–10) and did not display cytotoxicity (Yin et al. 2016).

Sponge-associated bacteria are known for their antimicrobial activity. For a detailed overview on their antimicrobial potential, please refer to the review by Santos-Gandelman and colleagues (2014), where a compilation of compounds, including those isolated from sponge-associated Pseudomonas spp., is provided. For instance, the strain P. putida Mm3, isolated from Mycale microsigmatosa, inhibited the growth of multidrug-resistant bacteria. The antimicrobial was resistant to NaOH and proteolytic enzymes, suggesting that its structure is devoid of a biologically active proteinaceous component or that the inhibition occurred upon production of organic acids (Marinho et al. 2009). Further, the study of Graça and colleagues (2013), which aimed to assess the antimicrobial potential of bacterial communities from the sponge Erylus discophorus, revealed that a sponge-associated Pseudomonas was active against B. subtilis, Vibrio harveyi, and Aliivibrio fischeri, as well as presented genes related to nonribosomal peptide synthetases (NRPS). The strains Pseudomonas fluorescens H40 and H41 and P. aeruginosa H51 isolated from Haliclona sp. were active against S. aureus, unveiling the presence of diketopiperazine cyclo-(LLeu-L-Pro), a compound with bactericidal effect toward S. aureus and P. aeruginosa and toxic to HEp-2 tumor cells (Santos et al. 2015). Later, it was demonstrated that genes related to the biosynthesis of the pyoverdine in P. fluorescens H41 were associated with its antimicrobial activity (Ye et al. 2015). Pseudomonas fluorescens H40 and H41 have been further investigated for their antibiofilm activity toward resistant Staphylococcus spp. isolated from canine skin. From 36 Staphylococcus strains tested, the CFS of P. fluorescens H40 and H41 dissociated the biofilm of 35 strains. The CFS from P. fluorescens H41 strongly dissociated (>75%) the mature biofilm of three strains of Staphylococcus pseudintermedius, being two multidrug resistant (Nunes et al. 2021).

The activity toward fungi has been explored in marine Pseudomonas. For example, the CFS of Pseudomonas sp. JK2, isolated from sea mud, inhibited the growth of phytopathogens, such as Aspergillus tamari, Fusarium solani, and Aspergillus fumigatus, by inducing cell wall thickening. However, the nature of the compound was not determined (Jin et al. 2013).

Antimicrobial and antilarval activities have been observed in a strain of Pseudomonas rhizosphaerae isolated from deep sea sediments. Using bioassay-guided assays, several compounds were obtained from the strain’s culture broth, including diketopiperazine and benzene-type metabolites. Among the compounds detected, some displayed activity toward Loktanella hongkongensis, Micrococcus luteus, Pseudoalteromonas piscicida, B. cereus, and Ruegeria sp. as well as inhibited larval settlement of the barnacle Balanus amphitrite (Qi et al. 2009). Examples of the antimicrobial potential of marine Pseudomonas are shown in Table 1.

Antifouling

The attachment of organisms to ship hulls and/or subsea equipment is known as marine biofouling, leading to significant economic losses, mainly due to increase in fuel consumption and maintenance costs. Therefore, antifouling strategies are urgently needed (Jin et al. 2022).

The extracts of the strain P. putida OM7 isolated from an octopus inhibited biofilm-forming strains of Alteromonas and Pseudomonas. Upon 5 h of treatment with either the intra or the extracellular extracts, a significant reduction of Alteromonas cell density was observed. Envisioning a potential application of the extracts as an antifouling agent, a coating was developed by mixing the bacterial extract with paint and applied to microscopic slides in vitro and to glass fiber plates in natural seawater. The slides were incubated with Alteromonas for 3 days, while the glass plates were kept submerged for 50 days. In both scenarios, biofilm formation on the tested surfaces was reduced when compared to the controls. In seawater, the adherence of organisms was reduced to 15.8 (intracellular extract) and 27.5% (extracellular extract) in comparison to the control (34.9%) (Viju et al. 2017).

The CFS of Pseudomonas sp. IV2006 inhibited the biofilm formation of Flavobacterium sp. II2003, known to produce a robust biofilm and to be one of the first colonizers of surfaces in the marine environment. Both strains were isolated from an intertidal mudflat biofilm sample. The CFS inhibited bacterial adhesion without killing or inhibiting bacterial growth. In static conditions, there was reduction of circa 70% of the Flavobacterium biofilm, while in dynamic conditions, a reduction of 60% in biofilm biovolume was reported, along with thickness reduction of 30%. By coating glass surfaces with the supernatant, the inhibition of biofilm formation was demonstrated. Efforts to characterize the antifouling agent revealed that it is likely of a proteinaceous nature, and it is relatively stable up to 50°C. Interestingly, the supernatant exerted antibiofilm activity with more than 50% reduction toward pathogens including Tenacibaculum maritimum, Vibrio lentus, S. aureus, Yersinia enterocolitica, and B. subtilis (Doghri et al. 2020).

Another example of the use of marine Pseudomonas strains in coatings to prevent biofouling was described by Kharchenko and colleagues (2012). The strain P. aeruginosa 1242 was isolated from the fouling microbiota of a brass plate and was selected for characterization based on its capacity to inhibit the growth of other microorganisms and to produce amylase, glycosidase, glycanase, and protease. An epoxy coating was developed using the immobilized cells of P. aeruginosa 1242, along with its enzymes and secondary metabolites. This coating was used to paint plastic panels and submerged in seawater of different climatic zones for up to 3 months, and results revealed that it significantly reduced adhesion and settlement of micro- and macrofoulers (Kharchenko et al. 2012).

Pigments

Natural pigments are currently employed in different areas, such as in food, cosmetic, and textile industries, as well as in medicine. The marine environment is a fruitful source of such compounds, which can mostly be found in invertebrates, algae, and microorganisms (Ye et al. 2019). When considering pigment-producing marine bacteria, Pseudomonas distinguish themselves by producing a variety of pigments with a broad array of functions, including antimicrobial, anti-cancer, and antioxidant (Nawaz et al. 2020).

For instance, melanin synthesis has been described in Pseudomonas. Melanin absorbs a wide range of radiation, which makes such pigments attractive to the cosmetic industry. The authors have described a melanin-producing Pseudomonas stutzeri strain isolated from red seaweed, whose production reached ∼6.7 g/l (Kumar et al. 2013). Another study demonstrated the production of pyomelanin by P. stutzeri BTCZ10 isolated from the Arabian Sea. Interestingly, when blended with commercial sunscreens, the pigment enhanced the sun protection factor (SPF; Kurian and Bhat 2018). Further, a Pseudomonas guineae strain isolated from seawater was also reported as a melanin-producing strain. The pigment exhibited antioxidant properties and could be produced in media containing cheap substrates, such as vegetable waste (Tarangini and Mishra 2013). Manirethan and colleagues (2018) have also demonstrated the ability of P. stutzeri HMGM-7, isolated from red seaweed, of producing a distinguished melanin adsorbent to heavy metals, including mercury, chromium, and lead. Later, the authors proposed the functionalization of this bacterial melanin for the efficient removal of arsenic from aqueous systems and rendering water fit for consumption. This process was achieved surpassing 99% arsenic removal efficiency until four cycles of adsorption and desorption (Manirethan et al. 2020).

Moreover, the species P. aeruginosa distinguishes itself by the number of reports on the characterization of its pigments. For example, the production of phenazine-1-carboxylic acid was described in P. aeruginosa strain GS-33 isolated from marine water collected in India (Patil et al. 2016). This pigment displayed activity toward the human skin melanoma cell line (SK-MEL-2) even at lower concentrations (<10 μg/ml). Further, it exhibited UV-B protection, enhanced the SPF of commercial sunscreens (10%–30% fold increase), and was non-toxic up to concentrations of 100 ppm. Additionally, a blue-green pigment produced by strain P. aeruginosa P1.S9, isolated from a marine sponge, was reported. The pigment’s crude extract exhibited antibacterial activity toward Gram-negative and Gram-positive bacteria, as well as toxicity toward cancer cell lines (MCF-12A and MCF-7) (Wahyudi et al. 2022).

Enzymes

Pseudomonas spp. are endowed with a remarkable enzymatic repertoire with a wide array of functions, spanning from antimicrobial resistance and virulence to biotechnological applications. Here, we have summarized the main reports on enzymes obtained from marine Pseudomonas strains and categorized them according to fields of application.

Biomedical applications

There is a need to improve current available treatments and to foster the development of innovative therapeutic strategies to combat existing and emerging diseases (Jagadeesan et al. 2019). In this scenario, enzymes emerge as compelling candidates for biomedical applications.

For instance, fibrinolytic enzymes are applied in the treatment of thrombosis, a cardiovascular disease marked by the aggregation of fibrin inside blood vessels. The disease is among the main causes of death worldwide, and current treatments are expensive and often present adverse side effects to patients (Altaf et al. 2021). Within this framework, the production of a fibrinolytic enzyme has been described in the strain P. aeruginosa KU1 isolated from marine sediments. By optimizing enzymatic production, a 3.25-fold increase in the production over prior to any optimization was reported, which is suitable for its large-scale production, reducing both time and costs involved in the production (Kumar et al. 2018). Later, this same research group reported the in vivo thrombolysis of the Pseudomonas-derived fibrinolytic enzyme. The enzyme was purified and reported as a metalloprotease with similarity with a serralysin-like alkaline protease from P. aeruginosa IFO3080. Both in silico and in vivo experiments revealed the enzyme as a promising candidate as a therapeutic agent, owing to its high affinity to bradykinin and its thrombolytic effect (Kumar et al. 2020).

Alginate lyases are regarded as interesting therapeutic agents, especially in the treatment of cystic fibrosis (CF). In the lung environment of CF patients, P. aeruginosa are found in biofilms and express a mucoid phenotype marked by the overproduction of alginate. Hence, efforts are directed toward the development of therapeutic agents that either target alginate production or lead to the enzymatic degradation of alginate, which can be achieved by alginate lyases (Blanco-Cabra et al. 2020). Further, alginate lyases can be applied to the production of alginate oligosaccharides, which are endowed with a milieu of biological activities, such as anti-inflammatory, antioxidant, and immunomodulatory (Chen et al. 2024). For instance, Li and colleagues (2011) have reported the purification and characterization of alginate oligosaccharides using an alginate lyase from Pseudomonas sp. HJZ216 isolated from brown seaweed.

L-asparaginases are hydrolases that catalyze the hydrolysis of acid asparagine into aspartic acid and ammonia and are studied for their role in the treatment of acute lymphoblastic leukemia, being most derived from E. coli and Erwinia chrysanthemi (Van-Trimpont et al. 2022). However, the search of novel enzymes from prokaryotic organisms paves the way for the discovery of unique properties that may offer advantages for therapeutic applications (Beckett and Gervais 2019). For example, an L-asparaginase derived from P. aeruginosa HR03 isolated from fish intestine demonstrated good thermal stability and was active in a wide range of pH. Even though cytotoxic assays were not conducted, the enzyme displays interesting properties that turn it into a candidate for pharmaceutical applications (Qeshmi et al. 2022).

The strain Pseudomonas oryzihabitans HUP022, isolated from deep sea, produces an esterase with high resistance to organic solvents, metal ions, surfactants, and NaCl with a proposed application in kinetic resolution of racemic ethyl 3-hydroxybutyrate and generation of ethyl (S)-3-hydroxybutyrate through direct hydrolysis reactions (Wang et al. 2016). After optimization, the yield of the final chiral product reached 87%. Ethyl 3-hydroxybutyrate is an intermediate for the synthesis of chiral drugs and chiral chemicals, which is an important step in the pharmaceutical industry owing to the different biological activities that enantiomers display. Typically, the synthesis of chiral chemicals involves harsh conditions, expensive cofactors, and toxic solvents. Thus, biocatalysis mediated by esterases emerges as a promising alternative to this scenario (Bornscheuer 2002).

Biodegradation of synthetic polymers

Plastics are recalcitrant pollutants and have quickly become a global issue. Hence, developing strategies to manage this kind of pollutant are needed, wherein microbial degradation arises as an interesting approach. Concerning Pseudomonas spp., we direct the reader to the review published by Wilkes and Aristilde (2017), where a thorough overview of the mechanisms by which Pseudomonas spp. metabolize plastic substrates and how they can be harnessed for bioremediation processes is provided. Here, however, we aim to summarize the main recent findings on polyester-degrading enzymes found exclusively in marine strains.

Bollinger and colleagues (2020b) have described a new polyester hydrolase produced by the strain Pseudomonas aestusnigri VGXO14^T, isolated from oil-polluted sand samples. Genomic analysis revealed the existence of a polyester hydrolase, PE-H, which could degrade polyethylene terephthalate (PET) and whose activity was significantly increased toward different substrates by the introduction of a single amino acid substitution, suggesting that protein engineering is an important step in optimizing PET hydrolysis on an industrial scale.

The strain Pseudomonas chengduensis BC1815, isolated from marine sediment, stood out for its ability to grow with PET as the sole carbon source (Shi et al. 2023). Genomic analysis revealed the presence of esterases, lipases, and α/β hydrolases, some of which membrane-bound and the others were secreted, suggesting that they are likely involved in PET degradation.

Additionally, Molitor and colleagues (2020) summarized screening strategies for the identification of polyesterases and applied these assays to demonstrate their production in strains belonging to the Pseudomonas pertucinogena lineage, including strains from marine habitats. The authors argued that this predominantly marine lineage holds great promise in the context of polyester hydrolysis owing to their psychrophilic and halophilic nature, as well as tolerance to heavy metals and hydrocarbonoclastic properties, thus underscoring the biotechnological potential of these bacteria, as supported by the aforementioned examples. A summary of the main enzymes discussed, their applications, and patents is disclosed in Table 2.

Bioremediation

Pseudomonas spp. are ideal candidates for bioremediation due to their versatility in using various carbon sources and their ability to reduce metal availability and toxicity. For instance, the strain P. stutzeri W228, isolated from sediment samples from Sepetiba Bay, Brazil, promoted lead absorption by the extracellular polymeric substances and the efflux of copper, making it suitable for removing these metals from contaminated environments (Coelho et al. 2020). Another example is strain P. putida SP1, isolated from seawater, which could volatilize 89% of the mercury it was exposed to and considered a candidate for the bioremediation of mercury-contaminated environments (Zhang et al. 2012). Further, the strain Pseudomonas pseudoalcaligenes S1, isolated from a mangrove area, could bioaccumulate mercury and adsorb this metal on the cell surface (Deng and Wang 2012).

Pollutants derived from myriad industries often reach the environment and cause threats to the local fauna and flora, as is the case of cadmium selenide (CdSe) nanocrystals. Strain P. fluorescens BA3SM1, isolated from a seaport, could counteract the toxicity of CdSe nanoparticles, turning it into a candidate for developing purification strategies to remediate contaminated waters (Poirier et al. 2016). Pseudomonas aeruginosa N6P6, isolated from seawater from a portuary region, was investigated for its naphthalene degradation potential. This strain was able to form biofilms under varied concentrations of naphthalene, and biofilm cultures degraded circa 99.4% of naphthalene in 20 h. The strain was able to degrade 64.3% of naphthalene in 24 h in soil microcosm experiments, suggesting the in situ application of this strain in naphthalene bioremediation (Kumari et al. 2021).

Pseudomonas spp. are also prolific producers of biosurfactants, which have been gaining growing interest in the bioremediation sector, especially because they enhance the dispersion and bioavailability of hydrophobic pollutants. Particularly, rhamnolipids are glycolipid biosurfactants typically produced by Pseudomonas that are endowed with a wide range of functions. For instance, P. aeruginosa ASW-4, isolated from marine intertidal sediments, produced a rhamnolipid that was applied to remove mercury and lead from marine sediments. At a critical micelle concentration of 43.73 mg/l, the treatment of sediment samples containing 520.32 mg/kg of lead and 13.15 mg/kg of mercury resulted in the removal of 62.50% of lead and 50.20% of mercury (Chen et al. 2021). Also, a strain of Pseudomonas guguanensis producing mono-rhamnolipids could remove n-hexadecane at a 77.2% removal rate on the sixth day after the bacterial inoculum when this hydrocarbon was supplemented to the bacterial culture. Interestingly, the dehydrated preparation could be stored at 35°C for up to 3 years, while still retaining biological activity. The fast degradation of hexadecane, diesel, and kerosene by the rhamnolipid, along with the non-pathogenic nature of P. guguanensis turn this organism into a potential candidate to remove n-alkanes from aquatic environments (Ramya Devi et al. 2018). Later, the same research group highlighted that this strain is a continuous producer of the rhamnolipid when fed with n-hexadecane as its sole carbon source and proposed the biochemical pathway for de novo synthesis of the mono-rhamnolipids, which could render higher (up to 2–3 folds) quantities for commercial use (Devi et al. 2019).

Pseudomonas aeruginosa AHV-KH10, isolated from contaminated seabed sediment, produced a rhamnolipid that could be applied in the bioremediation of diesel-contaminated sediments in saline environments. The biosurfactant demonstrated a diesel biodegradation rate of 70% for the initial concentration of 1000 mg/kg after 35 days. A biodegradation experiment was conducted in an unwashed sample of diesel-contaminated sediment for 4 months. When compared to the control, the Pseudomonas-derived rhamnolipid demonstrated 31.4% diesel removal efficiency, underscoring it as a candidate in bioaugmentation practices (Pourfadakari et al. 2021). Further, strain Pseudomonas sp. sp48, isolated from oil-contaminated water, could degrade oil, phenol, naphthalene, and pentadecane. The authors have provided a statistical optimization of the biodegradation process, rendering an 89% removal efficiency (Farag et al. 2018).

The di-rhamnolipids produced by Pseudomonas sp. S1WB, isolated from oil-contaminated water, were highly stable in terms of surface activity and emulsification when exposed to distinct temperatures, pH, and NaCl concentrations and were associated with petroleum degradation (Phulpoto et al. 2022). The strain P. aeruginosa NAPH6, isolated from seawater from a fishing harbor, degraded naphthalene, and crude oil under saline conditions. This strain also synthesized rhamnolipids on inexpensive carbon sources such as frying oil. The rhamnolipid was active on a wide range of temperatures, pH, and salinities and proved to be more efficient in hydrocarbon remobilization than synthetic surfactants (Hentati et al.2021).

A consortium of Pseudomonas isolated from Indian coastal waters was regarded as a candidate for the degradation of tributyltin (TBT), a biocide used in many antifouling paints. The five species included P. pseudoalcaligenes, P. stutzeri, P. mendocina, P. putida, and P. balearica, which were able to secrete high amounts of rhamnolipids and extracellular proteins. TBT degradation was proposed as a two-step process, in which dispersion of TBT in the aqueous phase is the first step, followed by cleavage by siderophores (Sampath et al. 2012).

Conclusions

Research provided over the last decades showcases the vast biotechnological potential of marine Pseudomonas (Fig. 3). However, such investigations are still in their infancy, as most of the available research comprises screening or preliminary characterization of bioactive molecules. Hence, few are the studies that propose and clearly indicate the potential application of the isolated compound. For instance, even though Pseudomonas is one of the leading genera in patent deposits, notably concerning bioremediation processes, a minority of those are derived from marine environments (Table 2) (Villela et al. 2019).

Recent advances in synthetic biology and genetic engineering will be crucial to optimize not only bioproducts and bioprocesses, but also bacterial strains themselves, as has been extensively investigated in the P. putida KT2440 strain (Nikel et al. 2014). Therefore, there is a window of opportunity to further investigate marine Pseudomonas, especially considering those isolated from underexplored marine environments, which could reveal compounds with unparalleled diversity.

Author contributions

Anna Luiza Bauer Canellas (Conceptualization [equal], Formal analysis [equal], Investigation [lead], Methodology [lead], Validation [equal], Visualization [lead], Writing – original draft [lead], Writing – review & editing [equal]), and Marinella Silva Laport (Conceptualization [equal], Data curation [lead], Formal analysis [equal], Funding acquisition [lead], Project administration [lead], Resources [lead], Supervision [lead], Validation [equal], Writing – review & editing [equal]).

Conflict of interest

None declared.

Funding

This work is supported by Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ, grant numbers: E-26/211.284/2021 andE-26/200.948/2021), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, grant numbers: 405020/2023-6 and 309158/2023-0) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, grant numbers: 88887.820714/2023-00, Financial Code 001).

Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

References