Errors in estimating reproductive parameters with macroscopic methods: a case study on the protogynous blacktip grouper Epinephelus fasciatus (Forsskål 1775).

A size-based, histological analysis of the reproductive life history of the blacktip grouper, Epinephelus fasciatus (Forsskål 1775), was conducted in Indonesia to evaluate the error rate associated with macroscopic reproductive analysis. Histological results indicated that E. fasciatus was protogynous with female L50 at 13.4 cm total length (LT) and a size at sexual transition of 22.0 cm LT. The weight-length relationship for the species was W = 0.011 L3.13. Overall sex ratios were significantly female biased, operational sex ratios were significantly male biased, and sex ratios of mature individuals varied predictably with length from female to male dominance as size increased. No significant relationship between length and batch fecundity was found. The population has a spawning period from February to August. Overall, 54.4% of macroscopic evaluations were incorrect compared to histological results. Of the errors, 14.8% were a failure to detect ovotestes, 12.7% were classifying non-gonadal tissue as ovary or testis, 12.2% were misclassifying sex, and 12.7% were misclassifying maturity status. However, the largest source of error (47.7%) was from misclassifying both sex and maturity status. Of these, 92.9% were macroscopically classified as immature females, but were histologically confirmed to be mature males. Compared to histological results, the only accurate macroscopic results were the absence of a sex-based difference in weight-length relationship and spawning seasonality estimated by a gonadosomatic index (February-June). The use of macroscopic methods to estimate reproductive life-history parameters for sex-changing reefes fish may introduce significant inaccuracies and misinterpretations. Of the parameters estimated by histological methods, size at maturity, size-specific sex ratios, and spawning seasonality have the greatest potential to inform local fishery management policy.

Physicochemical variables influencing the abundance of Atlantic goliath grouper juveniles (Epinephelus itajara) in a tropical Brazilian mangrove habitat.

The purpose of this study was to investigate the relationships between the abundance and occurrence of juvenile Atlantic goliath grouper and physicochemical variables in a tropical mangrove ecosystem, employing a non-lethal methodology. Juvenile Atlantic goliath grouper individuals were captured biweekly, using blue crab traps, across three locations within the São Mateus estuary (18°36'S, 39°44'W), from August 2021 to July 2022. Simultaneously, the physicochemical water variables were assessed during each trap inspection. The physicochemical water variables influence on the abundance and occurrence of Atlantic goliath grouper juveniles were assessed using the Zero-Inflated Binomial (ZIB) and the Poisson (ZIP) generalized linear mixed-effects models (GLMMs). A total of 66 Atlantic goliath grouper juveniles were sampled (Pontal do Sul = 50, Lagoa Creek = 13, and Pitangueiras Creek = 3), ranging from 32 to 319 mm in total length. The best model fitted to presence-absence data (AIC = 278.6) consisted of location, depth, and salinity, with only location having a significant influence on the likelihood of species occurrence. Regarding abundance data, the best model (AIC = 336.4) included location, depth, and turbidity, with only location and depth showing a significant impact on abundance variation, with lower abundance at deeper areas. Although our results indicate a weak significant influence of physicochemical variables on juvenile Atlantic goliath grouper distribution, it is imperative to continue this monitoring effort, considering that additional data may be required to gain a deeper understanding of these patterns.

Grouping groupers in the Mediterranean: Ecological baselines revealed by ancient proteins.

Marine historical ecology provides a means to establish baselines to inform current fisheries management. Groupers (Epinephelidae) are key species for fisheries in the Mediterranean, which have been heavily overfished. Species abundance and distribution prior to the 20th century in the Mediterranean remains poorly known. To reconstruct the past biogeography of Mediterranean groupers, we investigated whether Zooarchaeology by Mass Spectrometry (ZooMS) can be used for identifying intra-genus grouper bones to species level. We discovered 22 novel, species-specific ZooMS biomarkers for groupers. Applying these biomarkers to Kinet Höyük, a Mediterranean archaeological site, demonstrated 4000 years of regional Epinephelus aeneus dominance and resiliency through millennia of fishing pressures, habitat degradation and climatic changes. Combining ZooMS identifications with catch size reconstructions revealed the Epinephelus aeneus capacity for growing 30 cm larger than hitherto documented, revising the maximum Total Length from 120 to 150 cm. Our results provide ecological baselines for a key Mediterranean fishery which could be leveraged to define and assess conservation targets.

Hydroclimatic influence on the trophic ecology of Atlantic goliath grouper juveniles (Epinephelus itajara) in a tropical estuary using non-lethal methodologies.

The Atlantic goliath grouper Epinephelus itajara is the largest species of groupers in the Atlantic Ocean, reaching over 2 m total length and 400 kg. It has an ontogenetic migration, with adults using nearshore and offshore marine areas and juveniles inhabiting estuarine/mangrove areas. Despite of its high ecological relevance and classification as a threatened species for several decades in the IUCN red lists, critical phases of its life cycle, like juveniles inhabiting estuaries, are still poorly investigated. In this study, we evaluated if hydroclimatic changes may affect the trophic ecology of juveniles in a tropical estuary. We described their diet composition, isotopic niche area, food assimilation, and trophic position of juveniles across seasons using two non-lethal techniques: stomach content analysis using a stomach flushing procedure and stable isotope analysis using a dermal biopsy. A total of 87 individuals with TL ranging from 38 to 331 mm had their stomachs analyzed, of which 31 had food content. We identified 11 prey items, shrimp being the most abundant prey group (IRI% = 88.4), followed by fish (6.6), blue crab (2.8), and crab (2.2). A total of 93 specimens had their muscle's carbon (δ13C) and nitrogen (δ15N) stable isotope ratios analyzed (38-332 mm TL). Average δ13C values showed statistically significant changes across seasons, but no significant changes were observed for δ15N. As initially predicted, isotopic niche size of juveniles changed consistently along the entire hydroclimatic cycle, reaching its peak at the end of the wet season. We also observed changes in the proportion of prey assimilation by juveniles between seasons. Shrimp (92.1%) was the most assimilated prey group during the late dry season, whereas tidal crabs (36.7%), blue crab (27.8%), and shrimp (25.1%) were the most important in the late wet season. Average trophic position of juveniles remained similar across wet (3.32 ± 0.10) and dry (3.23 ± 0.11) seasons. These findings provide important information to help decision makers to promote more efficient management and conservation legislations to protect early life stages of the Atlantic goliath grouper in tropical estuaries.

Coastal upwelling influences population structure of dusky grouper Epinephelus marginatus: An integrative approach based on otolith chemistry and muscle stable isotopes.

The dusky grouper (Epinephelus marginatus) is an overfished and threatened fish species with coastal distribution. In the Southwestern Atlantic, it occurs across a broad region influenced by two major oceanographic features: the Cabo Frio (23°S) and the Cabo Santa Marta (28°S) upwelling systems. Along the Brazilian coast, the species may present continuous or discrete populations, depending on the methodological approach used. In this study we combined otolith chemistry and muscle stable isotope analyses to examine the population structure of dusky groupers and its association with the two upwelling systems. Fish were collected in shallow coastal waters of the Southwest Atlantic Ocean, covering the southeastern and southern Brazilian coasts, among Macaé (22°S), Santos (24°S), Florianópolis (27°S), and in Rio Grande (32°S). The results show three statistically well-separated population groups along the region. We named these population groups as North (north of Cabo Frio); Center (between upwelling regions); and South (south of the Cabo Santa Marta system). Our findings allow to suggest that the upwelling systems may influence the distribution of E. marginatus stocks along the Brazilian south-western coast, even though a causal effect may not be attributed at this point. Overall, this combined approach, leveraging information from distinct natural tags, and reflecting variability of water chemistry and food webs with latitude, allowed us to enhance our understanding on how major upwelling systems influence the structuring of fish populations along the southwestern Atlantic Ocean.

Age-based life history of three groupers in the southern Gulf of Mexico.

The three most important grouper fishery species in southern Gulf of Mexico, Epinephelus morio, Mycteroperca bonaci and Mycteroperca microlepis, do not have life history parameters data reported such as growth, age and natural mortality essential to management. Individuals from the three species were captured in the Campeche Bank (CB) at different times from 1988 to 2001. Sagittae were thin-sectioned to count annuli: n = 1081 for E. morio, n = 920 for M. bonaci and n = 251 for M. microlepis. Otoliths from the three species exhibited successive opaque and translucent bands deposited with an annual frequency, occurring between January and April for E. morio and M. bonaci, and between January and March for M. microlepis. Estimated age in the three grouper species ranged from 0+ to 20 years. The most frequent ages were 7-10 years for E. morio and 8-12 years for M. bonaci and M. microlepis. The von Bertallanfy Growth Function model parameters (L∞ , K and t0 ) were consistent with reported growth patterns for Epinephelidae family species, as established by growth performance Ф' (near 3). The CB groupers appear to reach maximum ages over 13 years with K values between 0.09 and 0.12 year-1 , indicating all three studied species displayed relatively slow growth rate and medium longevity. Lifespan values ranged from 13 to 33 years for E. morio, from 18 to 30 years for M. bonaci and from 17 to 33 years for M. microlepis. A50 was 6.1 years for E. morio and 5.5 years for both M. bonaci and M. microlepis. Age at which all females were mature was 8 years for M. bonaci and 10 years for E. morio and M. microlepis. P50 was 9.3 years for E. morio, 13.6 years for M. bonaci and 11.6 years for M. microlepis. Age at which 100% of individuals were male was 13 years for E. morio, 19 years for M. bonaci and 15 years for M. microlepis. Values of M at age exhibited an exponential decrease with age for all three species, but overall values were 0.13 year-1 for E. morio, 0.14 year-1 for M. bonaci and 0.17 year-1 for M. microlepis. For the three species size at age varied by geographical distribution area, but particularly for E. morio the discrepancy in average size between published studies may be the consequence of the fishing effort exerted on the stock and/or the lack of regulation for the fishery prior to 2001; after this stock biomass fell below the reference limit point and was declared overexploited. Resource managers could use the information provided in this study as a reference, representing pre-overexploitation values, to evaluate the current status of these populations and ensure sustainable management actions for the CB grouper fisheries.

Comparative Analysis of Four Complete Mitochondrial Genomes of Epinephelidae (Perciformes).

Groupers are commercial, mainly reef-associated fishes, classified in the family Epinephelidae (Perciformes). This study first sequenced the complete mitogenomes of Cephalopholis leopardus, Cephalopholis spiloparaea, Epinephelus amblycephalus, and Epinephelus hexagonatus. The lengths of the four Epinephelidae mitogenomes ranged from 16,585 base pair (bp) to 16,872 bp with the typical gene order. All tRNA genes had a typical cloverleaf structure, except the tRNA-Ser (AGY) gene which was lacking the entire dihydrouridine arm. The ratio of nonsynonymous substitution (Ka) and synonymous substitution (Ks) indicated that four groupers were suffering a purifying selection. Phylogenetic relationships were reconstructed by Bayesian inference (BI) and maximum likelihood (ML) methods based on all mitogenomic data of 41 groupers and 2 outgroups. The identical topologies result with high support values showed that Cephalopholis and Epinephelus are not monophyletic genera. Anyperodon and Cromileptes clustered to Epinephelus. Aethaloperca rogaa and Cephalopholis argus assembled a clad. Cephalopholis leopardus, C. spiloparaea, and Cephalopholis miniata are also in a clade. Epinephelushexagonatus is close to Epinephelus tauvina and Epinephelus merra, and E. amblycephalus is a sister group with Epinephelus stictus. More mitogenomic data from Epinephelidae species are essential to understand its taxonomic status with the family Serranidae.

Essential (Cu, Zn) and nonessential (Pb, Cd) metals in the muscle of leopard groupers (Mycteroperca rosacea) from a mining port in the Gulf of California, Mexico: human health risk assessment.

Mining activities are a current environmental issue due to heavy metal release and subsequent metal uptake by organisms. In this study, we quantified the concentrations of essential (Cu, Zn) and toxic (Cd, Pb) elements in the muscle of 248 leopard groupers, Mycteroperca rosacea, captured by spearfishing and free diving close to a mining district in the Gulf of California during 2014-2015. We analysed metals using high-resolution inductively coupled plasma source mass spectrometry (HR-ICP-MS). We analysed metal concentrations by fish size, sex, maturity, season, year and risk factor for human consumption. The results indicated common levels of essential elements (Cu: 11 ± 34.3 µg/g, Zn: 377 ± 1390 µg/g) in comparison with toxic elements (Cd: 0.06 ± 0.1 µg/g, Pb: 0.98 ± 1.5 µg/g). Cadmium was within the permissible limit of Mexican standards (0.5 µg/g), but lead content bordered its limit (1.0 µg/g). Heavy metal concentrations were comparable between males and females. Metal variations were not significantly correlated with sex, maturity, season or year (p > 0.05). The evaluation of benefits (daily mineral intake) and risks (target hazard quotients) to health indicated that these fish did not represent a risk of adverse effects to consumers within worldwide limits, while the nutritional benefits were high.

Age, growth and reproductive life-history characteristics infer a high population productivity for the sustainably fished protogynous hermaphroditic yellowspotted rockcod (Epinephelus areolatus) in north-western Australia.

The yellowspotted rockcod, Epinephelus areolatus, is a small-sized grouper that is widely distributed throughout the Indo-Pacific, where it forms a valuable component of the harvest derived from multispecies fisheries along continental and insular shelves. Samples of E. areolatus were collected from 2012 to 2018 from commercial catches and research surveys in the Kimberley, Pilbara and Gascoyne regions of north-western Australia to improve the understanding of the life history, inherent vulnerability and stock status of this species. Histological analysis of gonads (n = 1889) determined that E. areolatus was a monandric protogynous hermaphrodite. Non-functional spermatogenic crypts were dispersed within the ovaries of 23% of mature functioning females; nonetheless, these crypts were not observed during the immature female phase. The length and age at which 50% of females matured were 266 mm total length (LT ) and 2.7 years, respectively. The spawning period was protracted over 10-12 months of the year with biannual peaks at the start of spring and autumn (i.e., September and March) when the photoperiod was at its mid-range (i.e., 12.1 h). Estimates of the lengths and ages at which 50% of E. areolatus change sex from female to male were very similar (i.e., <5% difference) between the Kimberley and Pilbara regions, i.e., L 50 sc of 364 and 349 mm LT and A 50 sc of 7.9 and 7.3 years, respectively. A maximum age of 19 years was observed in all three regions, but there was significant regional variation in growth. These variations in growth were not correlated with latitude; instead a parabolic relationship was evident, where the smallest mean length-at-age and fastest growth rates (k) occurred in the mid-latitudes of the Pilbara region. In the Kimberley and Pilbara regions, individuals were not fully selected by commercial fish traps until 5-6 years of age, hence, several years after reaching maturity. These life-history characteristics infer a high population productivity, which underpins the sustainable harvest of this species, despite comprising the largest catches of all epinephelids in the multispecies tropical fisheries across north-western Australia.

Epinephelus tankahkeei, a new species of grouper (Teleostei, Perciformes, Epinephelidae) from the South China Sea.

A new species of grouper, Epinephelus tankahkeei sp. nov. is described from the South China Sea based on examination of morphological and molecular characteristics. This new species has been treated as, and is similar to, its congener E. chlorostigma. Epinephelus tankahkeei sp. nov. can be distinguished from E. chlorostigma by the following combination of characters: a convex anal fin; closer dark spots on the body; a lack of dark spots on the abdomen, cheek, and pectoral fin; the absence of a clear posterior white margin on the caudal fin. Molecular analyses of the mitochondrial COI sequence variation, genetic distances, and a phylogeny, all highly support E. tankahkeei sp. nov. as a distinct species. A key to E. tankahkeei sp. nov. and its most closely related species is provided.

Life-history strategies of the rock hind grouper Epinephelus adscensionis at Ascension Island.

Epinephelus adscensionis sampled from Ascension Island, South Atlantic Ocean, exhibits distinct life-history traits, including larger maximum size and size at sexual maturity than previous studies have demonstrated for this species in other locations. Otolith analysis yielded a maximum estimated age of 25 years, with calculated von Bertalanffy growth parameters of: L∞ = 55·14, K = 0·19, t0 = -0·88. Monthly gonad staging and analysis of gonad-somatic index (IG ) provide evidence for spawning from July to November with an IG peak in August (austral winter), during which time somatic growth is also suppressed. Observed patterns of sexual development were supportive of protogyny, although further work is needed to confirm this. Mean size at sexual maturity for females was 28·9 cm total length (LT ; 95% C.I. 27·1-30·7 cm) and no females were found >12 years and 48·0 cm LT , whereas all confirmed males sampled were mature, >35·1 cm LT with an age range from 3 to 18 years. The modelled size at which 50% of individuals were male was 41·8 cm (95% C.I. 40·4-43·2 cm). As far as is known, this study represents the first comprehensive investigation into the growth and reproduction of E. adscensionis at its type locality of Ascension Island and suggests that the population may be affected less by fisheries than elsewhere in its range. Nevertheless, improved regulation of the recreational fishery and sustained monitoring of abundance, length frequencies and life-history parameters are needed to inform long-term management measures, which could include the creation of marine reserves, size or temporal catch limits and stricter export controls.

Complete mitochondrial genome of the palemargin grouper Epinephelus bontoides (Pisces: Perciformes).

The complete mitogenome of the palemargin grouper, Epinephelus bontoides, was presented in this study. This mitochondrial genome consists of 16 903 bp, follow the typical gene arrangement with 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and a non-coding control region (CR). The overall base composition was A, 28.7%; G, 16.1%; C, 28.0%; and T, 27.2%. The control region was 1200 bp in length, which located between tRNAPro and tRNAPhe, rich in A + T (69.2%) content. Based on the Neighbor-Joining (NJ) tree, E. bontoides was grouped with E. trimaculatus, E. quoyanus, E. areolatus, and E. bleekeri, and then combined with E. merra formed a clade. This complete mitogenome of E. bontoides can provide essential phylogenetic information of Epinephelus.

Mercury concentrations in dusky grouper Epinephelus marginatus in littoral and neritic habitats along the Southern Brazilian coast.

Our study incorporated a comprehensive suite of parameters (i.e., body size, age, diet and trophic position) to investigate mercury concentration in dusky groupers Epinephelus marginatus. This study was carried out in rocky bottoms in littoral and neritic habitats along the Southern Brazilian coast. We also determined spatial variation in mercury concentrations in individuals inhabiting both zones, which may provide insights into how dietary differences or potential pollution sources affect bioaccumulation. A total of 244 dusky groupers was analyzed to determine total mercury concentrations. Our study revealed that when considering similar body sizes, individuals inhabiting littoral rocky habitats had higher concentrations of mercury probably due to proximity to pollution sources associated with human activities in the estuary and its drainage basin. Furthermore, large individuals (>650mm and >8years old) showed mercury contamination levels that are potentially harmful for this endangered fish species and above the acceptable limits for human consumption.

Evidence for spawning aggregations of the endangered Atlantic goliath grouper Epinephelus itajara in southern Brazil.

In this study, seasonal numerical abundance of the critically endangered Atlantic goliath grouper Epinephelus itajara was estimated by conducting scuba dive surveys and calculating sightings-per-unit-effort (SPUE) at three sites in southern Brazil. Seasonal differences in size and reproductive condition of captured or confiscated specimens were compared. The SPUE differed significantly with season, increasing in late spring and peaking during the austral summer months. A significant effect was observed in the number of fish relative to the lunar cycle. All females sampled during the summer were spawning capable, while all those sampled during other seasons were either regressing or regenerating. What these data strongly infer is that the E. itajara spawning aggregation sites have been located in the southern state of Paraná and the northern state of Santa Catarina and summer is the most likely spawning season. Size frequency distributions, abundance and reproductive state were estimated and correlated with environmental variables.

Complete mitochondrial genome and the phylogenetic position of the brown-spotted grouper Epinephelus chlorostigma (Perciformes: Epinephelidae).

The complete mitogenome of the brown-spotted grouper Epinephelus chlorostigma was determined in this study. It is 16,894 bp in length, containing 37 genes with the typical order and transcriptional direction in the vertebrates. The overall nucleotide composition is: 28.7% A; 28.0% C; 16.3% G and 27.0% T. The lengths of 22 transfer RNA (tRNA) genes range from 67 (tRNA-Cys) to 76 (tRNA-Leu1) bp. Two start codons (GTG and ATG) and two stop codons (TAG and TAA/T) were found in the protein-coding genes. The nodes of phylogenetic tree were strongly supported based on the complete mitogenomes of available groupers. The phylogenetic results suggested E. chlorostigma is the closest to E. areolatus within the genus Epinephelus.

Restricted grouper reproductive migrations support community-based management.

Conservation commonly requires trade-offs between social and ecological goals. For tropical small-scale fisheries, spatial scales of socially appropriate management are generally small-the median no-take locally managed marine area (LMMA) area throughout the Pacific is less than 1 km(2). This is of particular concern for large coral reef fishes, such as many species of grouper, which migrate to aggregations to spawn. Current data suggest that the catchment areas (i.e. total area from which individuals are drawn) of such aggregations are at spatial scales that preclude effective community-based management with no-take LMMAs. We used acoustic telemetry and tag-returns to examine reproductive migrations and catchment areas of the grouper Epinephelus fuscoguttatus at a spawning aggregation in Papua New Guinea. Protection of the resultant catchment area of approximately 16 km(2) using a no-take LMMA is socially untenable here and throughout much of the Pacific region. However, we found that spawning migrations were skewed towards shorter distances. Consequently, expanding the current 0.2 km(2) no-take LMMA to 1-2 km(2) would protect approximately 30-50% of the spawning population throughout the non-spawning season. Contrasting with current knowledge, our results demonstrate that species with moderate reproductive migrations can be managed at scales congruous with spatially restricted management tools.

Primary male development of two sequentially hermaphroditic groupers, Epinephelus akaara and Epinephelus awoara (Perciformes: Epinephelidae).

Gonad ontogeny of the Hong Kong grouper Epinephelus akaara (a bi-directional sex changer) and the yellow grouper Epinephelus awoara (a protogynous hermaphrodite) was examined for the first time from post-larval phase until first sexual maturation, by histology. Approximately 20 specimens of each species were collected randomly every 2-7 weeks from rearing tanks with natural sea water and temperature between June 2013 and June 2014. The paired gonadal primordia (GP) were observed at 6 weeks after hatching (wah) for both species; however, gonia were first observed in GP at 16 wah for E. akaara and at 8 wah for E. awoara. The timings for the appearance of primary-growth stage oocytes (O1) and the completion of ovarian lumen (OL) varied; both at 27 wah for E. akaara, and at 18 and 23 wah for E. awoara respectively. A bisexual-phase gonad with an OL, O1 and scattered spermatogenic cysts (SC) was observed at 27-29 wah for both E. akaara and E. awoara. Sexual differentiation was subsequently observed from the bisexual-phase gonad at 34 wah for E. akaara, and 41 wah for E. awoara, with the appearance of cortical-alveolus stage oocytes (O2) for developing female and the proliferation of SC for developing primary male (i.e. from juvenile directly). Ovaries of mature females contained the vitellogenic stage oocytes (O3) and scattered SC; testes of mature primary males had sperm in sperm sinuses within the gonadal wall and remained O1. Minimum age of first sexual maturation for both female and primary male of E. akaara was at 41 wah; minimum total length (LT ) of female (143 mm) was larger than that of primary male (137 mm L(T)). Minimum age and size of first sexual maturation for female of E. awoara (47 wah and 149 mm L(T), respectively) were larger than those of E. akaara. Developing primary males of E. awoara were found at 41-58 wah, however, mature males were not observed, indicating inconsistency in first sexual maturation for E. awoara. This study provided strong evidences of primary male pathway in E. akaara and E. awoara; the latter is confirmed to be diandric.

Complete mitochondrial genome of the sixblotch hind Cephalopholis sexmaculata (Pisces: Perciformes).

The complete mitogenome of the sixblotch hind, Cephalopholis sexmaculata was presented in this study. This mitochondrial genome consists of 16,589 bp, with 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and a noncoding control region (CR), and its gene arrangement is identical to most vertebrates. The overall base composition of the heavy strand is A, 29.35%; G, 16.08%; C, 28.56%; and T, 26.01%. The COI gene started with GTG codon and the ATP6 gene started with CTG codon. The complete mitogenomic data may provide informative for further phylogenetic approach of species of Cephalopholis and related genera belong to the Epinephelidae groupers.

First record of a spawning aggregation for the tropical eastern Pacific endemic grouper Mycteroperca olfax in the Galapagos Marine Reserve.

This study provides direct and indirect evidence of temporally and spatially consistent spawning aggregations for the grouper Mycteroperca olfax. Recently reported declines in population numbers, probably related to the direct targeting of aggregations by artisanal fishermen, highlight the urgent need for species-specific management actions in the Galapagos Marine Reserve, such as minimum and maximum landing sizes, and the importance of protecting key aggregation sites with the declaration of no-take areas and the establishment of total fishing bans during the reproductive season.

Spawning aggregations of three protogynous groupers in the southern Gulf of Mexico.

Spawning aggregations of red hind Epinephelus guttatus, tiger grouper Mycteroperca tigris and yellowfin grouper Mycteroperca venenosa were identified at two coral-reef systems: Arrecife Alacranes (emergent bank reef) and Bajos del Norte (submerged bank reef) on the continental shelf of the Yucatan Peninsula (Campeche Bank), Mexico. At both reefs, E. guttatus forms large spawning aggregations between February and March. At Bajos del Norte, M. tigris reproduces in a small, low-density aggregation in May, while M. venenosa aggregates at high densities for spawning between March and May. Multi-species use of an aggregation site by E. guttatus and M. venenosa was observed at Bajos del Norte. The identified spawning aggregations are apparently stable in location over time, and all three species were commonly observed to spawn within 1 week following the full moon. Development and survival of the larvae spawned in these aggregations are probably aided by a seasonal (spring-summer) upwelling in the north-east Campeche Bank. A permanent area closure at Bajos del Norte, currently outside any specific fisheries management area or regulations, would provide protection needed for the spawning aggregations of these three species.