Checklist of marine macroalgae in two contiguous Marine Protected Areas in the south-western Atlantic

Abstract Background The Costa das Algas Environmental Protection Area (EPA) and the Santa Cruz Wildlife Refuge (WR), located in the Espírito Santo Continental Shelf, Brazil, are outstanding marine protected areas due to their high biodiversity, particularly of macroalgae. Together, these two relatively small protected areas (1,150 and 177 km2, respectively) harbour about a quarter of all macroalgal species recorded in Brazil. The checklist presented herein updates the algal flora of these two protected areas with data obtained until 2019. Two hundred and sixty-five macroalgal taxa were recorded, most of which with vouchers. Checklists based on the collections of each protected area were published on: "Catálogo de Plantas das Unidades de Conservação do Brasil" (https://catalogo-ucs-brasil.jbrj.gov.br/). New information Besides specimens collected between 2018 and 2019, the algal flora presented herein includes previous records from different Brazilian herbaria (e.g., SP, SPF, ALCB). Herbaria records may include species that do not occur in intertidal reefs (e.g., Laminaria). Overall, 249 macroalgal taxa and one marine angiosperm were recorded in the Costa das Algas EPA (87 new records) and 136 macroalgal taxa and one marine angiosperm in the Santa Cruz WR (46 new records). All taxa are native to Brazil and nine are endemic to Brazil. Our results provide a taxonomic foundation to support management, long-term monitoring and conservation in these protected areas.

The checklist presented herein updates the algal flora of these two protected areas with data obtained until 2019.Two hundred and sixty-five macroalgal taxa were recorded, most of which with vouchers.Checklists based on the collections of each protected area were ‡

Introduction
In coastal zones, benthic macroalgae are important primary producers that provide habitat and food for several organisms and multiple benefits for people.They contribute for food, fisheries support, nutrient cycling, coastal protection, water quality and carbon storage (Duffy et al. 2019).However, they are often threatened by several human activities, such as coastal development, urban wastes, invasive species, climate change, oil and gas and industrial activities (Duffy et al. 2019, Hanley et al. 2024).
The Espírito Santo State (ES) encompasses one of Brazil's highest species richness of marine macroalgae, with records of about 400 species (Guimarães 2003, Flora e Funga do Brasil 2023).Its shoreline is composed of beaches, bays, inlets, mangroves, estuaries and lateritic reefs, while its continental shelf harbours a variety of sedimented bottom and rhodolith beds (Bastos et al. 2015, Quaresma et al. 2015), both punctuated by biogenic reefs and other mesoscale benthic features (Holz et al. 2020, Oliveira et al. 2023).The diversity of structurally complex hard bottom habitats contributes to the great intertidal and subtidal diversity of algae (Guimarães 2003, Amado-Filho et al. 2010, Holz et al. 2020).Moreover, the study region is situated in a transitional zone between the tropical and warm temperate phycogeographical provinces (Horta et al. 2001), allowing for the co-occurrence of species with affinities for both warm and cold waters (Horta et al. 2001).
The multiple-use Costa das Algas EPA and the no-take Santa Cruz WR are contiguous and form part of UNESCO's Mata Atlântica Biosphere Reserve.Established in 2010 by Brazil's Federal Government, these marine protected areas (MPAs) aim at the protection of marine biodiversity and management of artisanal fisheries, and were important milestones for the protection of rhodolith beds against large-scale exploitation for the production of agriculture fertilisers [IBAMA (Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis) 2006].Both areas have great ecological and socioeconomic importance and protect critical habitats that ensure coastal zone protection against erosion and the sustainability of artisanal fisheries, the latter being a major economic activity in the region [ICMBio (Instituto Chico Mendes de Conservação da Biodiversidade) 2023a].Moreover, they are major tourism destinations, especially during the summer.These areas are faced with several threats, including the contamination by heavy metals from mining wastes transported by the Doce river, with highly increased concerns after the collapse of the Fundão Dam in November 2015 [ICMBio (Instituto Chico Mendes de Conservação da Biodiversidade) 2023b].Oliveira-Filho (1977) was the first to highlight the outstanding algal diversity in ES.However, inventories are limited to a few publications (Oliveira-Filho 1969, Oliveira-Filho 1976, Oliveira-Filho 1976, Guimarães 2006, Scherner et al. 2013) and some data was only available in grey literature (Crispino 2000, Barata 2004, Carvalho 2013).Furthermore, most inventories were carried out before 2010 and did not include geographic coordinates or vouchers.In an effort to reduce this knowledge gap, this study aimed at providing an updated checklist of macroalgae for the EPA and the WR.

Study areas
This study focused on macroalgae associated with intertidal reefs in Costa das Algas EPA and Santa Cruz WR, in the Espirito Santo Continental Shelf (ESCS, Oliveira et al. (2020)), south-eastern Brazil.The framework of these reefs, which dwell under a micro-tidal regime and strong fluvial influence, is composed of lateritic rocks interspaced with calcareous formations (Mazzucco et al. 2020).On the Espírito Santo coast, temperatures range from 25 to 28ºC in the summer and from 22.5 to 24.5ºC in the winter (Instituto Estadual de Meio Ambiente e Recursos Hídricos 2014).Upwelling associated to the intrusion of the South Atlantic Central Water impacts the ESCS, lowering the temperature during short periods between October and March (Guimarães 2003).
Checklists of species with vouchers in the main national herbaria were published on the "Catálogo de Plantas das Unidades de Conservação do Brasil" (https://catalogo-ucsbrasil.jbrj.gov.br/) for both MPAs.Here, we report 30 additional species recorded in our field surveys, but not found in the herbaria.Their abundance was assessed according to the frequencies of occurrence in the eight studied sites and four different seasons in the field surveys (2018)(2019), as follows: very common (VC = 28-32 occurrences); common (C = 21-27 occurrences); frequent (F = 3-10 occurrences) and rare (R = 1 or 2 occurrences).Coralline algae were often identified only at the genus level, since species level identification of these algae usually requires molecular analysis.
Statistical analysis: Diversity of macroalgae of both MPAs was summarised at family and genus level, as well as endemism and status as native or non-native, based on Flora e Funga do Brasil (2023).

Geographic coverage
Description: The Costa das Algas EPA covers 1,150 km , with about 96% of its area in the marine environment and the remaining in restingas, beaches and mangroves (Fig. 2).The Santa Cruz WR covers 177 km , 98% in the marine environment.Altogether, these MPAs span about 30 km of shoreline.

Analysis Taxonomy coverage
Two hundred and sixty-five macroalgal taxa were recorded in the protected areas, including four varieties, one subspecies and one marine angiosperm (Table 1, Suppl.material 1, Suppl.material 2).Costa das Algas EPA (250 taxa) was richer than the Santa Cruz WR (137 taxa) due to the combination of lower sampling effort (only two sampling sites) and smaller area of the latter (Fig. 3).

:
This inventory was carried out within the scope of two projects: 1) T1) Aquatic Biodiversity Monitoring Program (PMBA-FEST, RENOVA); 2) Long-Term Ecological Research Program (PELD-Abrolhos/CNPq); Personnel: The research is Cláudia S Karez Post-Doctoral/CNPq project in the Jardim Botânico do Rio de Janeiro (JBRJ), supervised by Leonardo T Salgado.Identifications were made under the supervision of José Marcos C Nunes (Universidade Federal da Bahia) and Ricardo G Bahia (JBRJ) for crustose coralline algae.Both project are coordinated by Rodrigo L Moura (Universidade Federal do Rio de Janeiro).Study area description: Sampling was carried out in the Espirito Santo Continental Shelf (ESCS, Oliveira et al. (2020)), south-eastern Brazil, in intertidal reefs within two contiguous marine protected areas, the Costa das Algas EPA (multiple-use) and the Santa Cruz WR (no-take).Funding: The inventory was funded by the Aquatic Biodiversity Monitoring Program (PMBA-FEST, RENOVA) and the Long-Term Ecological Research Program (PELD-Abrolhos/ CNPq).Brazil's Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) has granted Research Productivity (JMCN, RLM, PSS, LTS) and Post-Doctoral (CSK) fellowships.We thank the "Núcleo de Gestão Integrada de Santa Cruz" -Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio) for research permits and background information.

Figure 1 .
Figure 1.Shallow reef benthic assemblages adjacent to the coastline dominated by algae in the Costa das Algas EPA and Santa Cruz WR (2018-2019).A Zoanthid (Palythoa sp.) and algae; B Halimeda sp.; C Vidalia obtusilosa; D Tricleocarpa cylindrica; Е Maerls formed by living monospecific CCA covering the sea bed associated with Dictyoperis delicatula; F Living rhodoliths surrounded by nodules covered by turf; G Living rhodoliths associated with Padina sp. and Sargassum sp.; H Collection of macroalgae in the field; Photos: (A-C) A. Bertoncini; Photos (D-H) F. Moraes.
Figure 2. Site location and views of the two studied MPAs.A Sampling sites in the multiple use Costa das Algas Environmental Protection Area (larger polygon) and in the no-take Santa Cruz Wildlife Reserve (smaller polygon); B Costa das Algas EPA; C Santa Cruz WR.Photos: F. Moraes.

Figure 3 .
Figure 3. Number of macroalgal taxa of Rhodophyta, Chlorophyta and Ochrophyta in the Costa das Algas EPA and the Santa Cruz WR (ES).

Costa das Algas EPA and Santa Cruz WR Phylum Chlorophyta/Ochrophyta/Rhodophyta/Tracheophyta Notes: Macroalgae recorded in the studied areas are presented in Table 1 PHYLUM/CLASS/ORDER/FAMILY/SPECIES Costa das Algas EPA Santa Cruz WR Period record Abundance CHLOROPHYTA CLASS ULVOPHYCEAE BRYOPSIDALES Bryopsidaceae
Bryopsis pennata J.V.Lamouroux 1809 X X PR Bryopsis plumosa (Hudson) C.Agardh 1823 X PR Caulerpaceae Caulerpa ambigua Okamura 1897 Checklist of marine macroalgae in two contiguous Marine Protected Areas ... Checklist of marine macroalgae in two contiguous Marine Protected Areas ...
(Scherner et al. 2013)herner et al. 2013mi, Ximenes et al. 201743 species (60% of all taxa) in the Costa das Algas EPA and 29 families, 49 genera and 68 species (53% of all taxa) in the Santa Cruz WR.Chlorophyta consisted of 15 families, 21 genera and 57 species (24% of all taxa) in the Costa das Algas EPA and 10 families, 14 genera and 30 species (22% of all taxa) in the Santa Cruz WR.Ochrophyta was composed of seven families, 16 genera and 39 species (16% of all taxa) in the Costa das Algas EPA and five families, 14 genera and 33 species (24% of all taxa) in the Santa Cruz WR.Laminaria abyssalis (Ochrophyta) and Halimeda jolyana (Chlorophyta)(Nunes 1998, Nunes 2005, Moura et al. 2015, Ximenes et al. 2017, Flora e Funga do Brasil 2023).One hundred and four species (40% of all taxa), recorded before the 2015 themining disaster, were also presented in our 2018-2019 surveys.There were 86 new occurrences (34% of all taxa in the MPA) for the Costa das Algas EPA and 45 new ones (33% of all taxa in the MPA) for the Santa Cruz WR in the 2018-2019 surveys.On the other hand, 64 previously recorded taxa (45 Rhodophyta, 14 Chlorophyta and 5 Ochrophyta) were not found in our surveys.Chlorophyta are smaller and filamentous forms, that tends to be more tolerant to changes in the environment [WFD-UKTAG (Water Framework Directive -United Kingdom Technical Advisory Group) 2009].The red algal order, Ceramiales, which tends to thrive under better environmental quality(Bermejo et al. 2012) encompassed the higher richness in the study area (66 species), both before and after the 2015 mining disaster, but 22 of the previously recorded species of this group were not found during the latter surveys.An inventory carried out in the 2000s (Guimarães 2003), compared the Ceramiales flora of Espírito Santo (115 species) with surveys carried ou 30 years earlier(Oliveira-Filho 1969)and recorded no differences in the algal composition.Ecological impacts of terrestrial nutrients, sediments and pollutants are likely to be species-specific(Díaz-Pulido et al. 2007), and may lead to insidious changes in the macroalgal assemblages.The two protected areas studied herein are historically degraded due to multiple chronic impacts(Costa et al. 2021, Scherner et al. 2013), but the contaminants from the 2015 mining disaster are especially concerning and deserve further monitoring efforts(Quaresma et al. 2021, Coppo et al.  2023).New records of algae after the mining disaster are due either to the increased sampling effort near the Doce river mouth or to taxonomic studies during the two last decades.Some of these newly recorded taxa are indeed recent description, and it should be also mentioned that amongst the 100 new records comprise rare species.Furthermore, there are new records amongst Dictyotales, Fucales and Corallinales, which are the most abundant algal groups and have an intricate taxonomy due to morphological variability associated to environmental conditions(Bahia 2014, González-Nieto et al. 2020).For example, in a recent molecular study, 10 species of Sargassum ( S. bermudense, S.Our new algal records do not mean environmental quality improvements in the Espirito Santo coast.Indeed, new algal records are frequent in degraded environments, which tend to receive adequate monitoring efforts after enviroments impacts.Examples include Santos(Oliveira and Qi 2003), Sepetiba (Amado-Filho et al. 2003)and Guanabara bays(De Paula  et al. 2020), as well as the vicinity of other urban centres(Scherner et al. 2013).Therefore, actual decreases in diversity are frequent despite larger checklists.For instance, Amado-Filho et al. (2003) recorded reduction of Rhodophyta richness, together with dominance of Sargassum spp.and Padina gymnospora under high metal concentrations in Sepetiba Bay (Brazil).Algal assemblages are excellent indicators of climate and anthropogenic impacts ( Mazzucco et al. 2020), Carneiro et al. 2023).
buxifolium, S. cymosum, S. filipendula, S. fluitans, S. furcatum, S. hystrix, S. natans, S. polyceratium and S. vulgare) were synonymised under S. cymosum(González-Nieto et al.  2020).Here, we did not use González-Nieto et al. (2020) taxonomic arrangement, as their analyses were largely based on data from the Caribbean with only one sequence from Brazil.and threatened MPAs is essential to guide management actions and restoration efforts, and the baseline data herein providesa relevant foundation for further assessments.