Deep-sea mining: using hyperbaric conditions to study the impact of sediment plumes in the subtidal clam Spisula solida.

With the growing interest to exploit mineral resources in the deep-sea, there is the need to establish guidelines and frameworks to support hazard and risk assessment schemes. The present study used a subtidal species of filter-feeding bivalve, the clam Spisula solida, as a proxy to better understand the impacts of sediment plumes in marine organisms under hyperbaric conditions. Four concentrations of suspended sediments (0 g/L, 1 g/L, 2 g/L, and 4 g/L) were used in a mixture with different grain sizes at 4 Bar for 96 h. Functional (filtration rate-FR) and biochemical endpoints (catalase-CAT, glutathione s-transferase-GST, and lipid peroxidation-LPO) were analyzed in the gonads, digestive gland, and gills of S. solida after a 96-h exposure at 4 Bar (the natural limit of the species vertical distribution). The FR showed a decreasing trend with the increasing sediment concentrations (significant effects at 2 and 4 g/L). Additionally, significant changes were observed for some of the tested oxidative stress biomarkers, which were concentration and tissue-dependent, i.e., CAT activity was significantly elevated in gills (1 g/L treatment), and GST was decreased in digestive gland (1 g/L treatment). Overall, the results show that suspended sediments, at 2 and 4 g/L, have negative functional impacts in the bivalve S. solida providing additional insights to improve hazard assessment of deep-sea mining. These findings represent a step forward to ensure the mitigation of the potential negative effects of deep-sea resource exploitation.

Morphological traits for allometric scaling of the European Sea Bass Dicentrarchus labrax (Linnaeus, 1758) from Southern Portugal population.

The present study aimed to determine the allometric scaling among a selection of morphological traits in European sea bass (Dicentrarchus labrax) to estimate fish body weight. A set of morphological traits (fish body weight, length, height, and width) were directly measured in 146 fish of a recirculating aquaculture system, with body weights ranging from 17.11 to 652.21 g. In addition, a collection of digital imagery of each anesthetized fish from the side and top views were used to estimate other traits (indirect measures). Multiple regression analysis and regression coefficients were calculated using all possible combinations of biometric data (predictors) to estimate fish body weight, applying different numerical fitting models (linear, log-linear, quadratic, exponential). The results showed that the best combination of traits for estimating fish body weight were fish body width, length and height, collected from direct measure (R2 = 0.995), for a log-linear model fitting, which revealed more accurate determinations than the most commonly used length-weight relationship. Nevertheless, other combinations of morphological traits and fitting models were also found to be suitable in successfully predict fish body weight, with variability ranging between 92.5% and 98.5%. For indirect measures, the best predictor was a combination of traits from top view (width, eye distance and area without fins) fitted with a log-linear function. These results comprise a relevant baseline in supporting the high potential of noninvasive methods to accurately follow the growth of European sea bass juveniles, recurring to imagery analysis of anesthetized fish. It has major potential applications in feeding consumption trials and fish growth models, as it allows for continuously following up fish growth under different experimental conditions without therein distress derived from manipulation.

A modelling framework to assess multiple metals impacts on marine food webs: Relevance for assessing the ecological implications of deep-sea mining based on a systematic review.

Industrial deep-sea mining will release plumes containing metals that may disperse over long distances; however, there is no general understanding of metal effects on marine ecosystems. Thus, we conducted a systematic review in search of models of metal effects on aquatic biota with the future perspective to support Environmental Risk Assessment (ERA) of deep-sea mining. According to results, the use of models to study metal effects is strongly biased towards freshwater species (83% freshwater versus 14% marine); Cu, Hg, Al, Ni, Pb, Cd and Zn are the best-studied metals, and most studies target few species rather than entire food webs. We argue that these limitations restrain ERA on marine ecosystems. To overcome this gap of knowledge, we suggest future research directions and propose a modelling framework to predict the effects of metals on marine food webs, which in our view is relevant for ERA of deep-sea mining.

Stressors of emerging concern in deep-sea environments: microplastics, pharmaceuticals, personal care products and deep-sea mining.

Although most deep-sea areas are remote in comparison to coastal zones, a growing body of literature indicates that many sensitive ecosystems could be under increased stress from anthropogenic sources. Among the multiple potential stressors, microplastics (MPs), pharmaceuticals and personal care products (PPCPs/PCPs) and the imminent start of commercial deep-sea mining have received increased attention. Here we review recent literature on these emerging stressors in deep-sea environments and discuss cumulative effects with climate change associated variables. Importantly, MPs and PPCPs have been detected in deep-sea waters, organisms and sediments, in some locations in comparable levels to coastal areas. The Atlantic Ocean and the Mediterranean Sea are the most studied areas and where higher levels of MPs and PPCPs have been detected. The paucity of data for most other deep-sea ecosystems indicates that many more locations are likely to be contaminated by these emerging stressors, but the absence of studies hampers a better assessment of the potential risk. The main knowledge gaps in the field are identified and discussed, and future research priorities are highlighted to improve hazard and risk assessment.

Boom and bust: Simulating the effects of climate change on the population dynamics of a global invader near the edge of its native range.

Despite the increasing awareness of climate change, few studies have used the Intergovernmental Panel on Climate Change (IPCC) scenarios to simulate the effects of climate change on estuarine populations of crustaceans. The objective of this study was to investigate the effects of temperature and salinity fluctuations on the population dynamics of the shore crab Carcinus maenas at the southern edge of its native range. To this end, a population dynamics model was developed based on experimental and literature data on the biology, ecology and physiology of the species. Results showed that the shore crab will be more affected by changes in temperature than in salinity. The parameter sensitivity analysis revealed that the larval phase of the species is the most sensitive stage of the shore crab life cycle. Three IPCC scenarios (SSP1-2.6, SSP2-4.5, and SSP3-8.5) were used to simulate the effects of temperature increase on the population of C. maenas in the near- (2040), mid- (2060), and long-term (2100). Two scenarios of drought conditions accompanied by the estimated salinity change were also simulated (10 % and 40 % drought). Results suggested that slight increases in temperature (up to 2 °C) lead to a strong increase on the density of C. maenas in the mid-term, while further temperature increases lead to a decline or local extinction of the shore crab population at the southern edge of its distribution range. Results indicated that a salinity increase in the estuary had a negative effect on the shore crab population. Given the importance of the species to temperate coastal ecosystems, both population increase and local extinction are likely to have significant impacts on estuarine communities and food webs, with unknown ecological and socioeconomic consequences.

Prioritizing the Effects of Emerging Contaminants on Estuarine Production under Global Warming Scenarios.

Due to non-linear interactions, the effects of contaminant mixtures on aquatic ecosystems are difficult to assess, especially under temperature rise that will likely exacerbate the complexity of the responses. Yet, under the current climatic crisis, assessing the effects of water contaminants and temperature is paramount to understanding the biological impacts of mixtures of stressors on aquatic ecosystems. Here, we use an ecosystem model followed by global sensitivity analysis (GSA) to prioritize the effects of four single emerging contaminants (ECs) and their mixture, combined with two temperature rise scenarios, on the biomass production of a NE Atlantic estuary. Scenarios ran for 10 years with a time-step of 0.1 days. The results indicate that macroinvertebrate biomass was significantly explained by the effect of each single EC and by their mixture but not by temperature. Globally, the most adverse effects were induced by two ECs and by the mixture of the four ECs, although the sensitivity of macroinvertebrates to the tested scenarios differed. Overall, the present approach is useful to prioritize the effects of stressors and assess the sensitivity of the different trophic groups within food webs, which may be of relevance to support decision making linked to the sustainable management of estuaries and other aquatic systems.

Trichoderma from Brazilian garlic and onion crop soils and description of two new species: Trichoderma azevedoi and Trichoderma peberdyi.

Fifty four Trichoderma strains were isolated from soil samples collected from garlic and onion crops in eight different sites in Brazil and were identified using phylogenetic analysis based on combined ITS region, tef1-α, cal, act and rpb2 sequences. The genetic variability of the recovered Trichoderma species was analysed by AFLP and their phenotypic variability determined using MALDI-TOF. The strain clusters from both typing techniques coincided with the taxonomic determinations made from phylogenetic analysis. The phylogenetic analysis showed the occurrence of Trichoderma asperellum, Trichoderma asperelloides, Trichoderma afroharzianum, Trichoderma hamatum, Trichoderma lentiforme, Trichoderma koningiopsis, Trichoderma longibrachiatum and Trichoderma erinaceum, in the soil samples. We also identified and describe two new Trichoderma species, both in the harzianum clade of section Pachybasium, which we have named Trichoderma azevedoi sp. nov. and Trichoderma peberdyi sp. nov. The examined strains of both T. azevedoi (three strains) and T. peberdyi (12 strains) display significant genotypic and phenotypic variability, but form monophyletic clades with strong bootstrap and posterior probability support and are morphologically distinct from their respective most closely related species.

MODELPlastics workshop - Modelling Ocean Plastic Litter in a Changing Climate: Gaps and future directions.

Here we summarize the overarching issues that emerged from a workshop held to discuss scientific challenges and future directions on the use of numerical models to predict the amount, distribution and effects of plastic pollution on marine ecosystems. The need for multi-disciplinary approaches, standardized protocols for plastic quantification and analyses, using realistic contaminant concentrations in laboratorial experiments and targeting early-life stages of marine organisms were pointed out as needs to improve data accuracy. Participants also enumerated a list of gaps that include, identification of indicator organisms of plastic contamination, selection of biomarkers and the role of extreme events on plastic dynamics. Responding to these gaps will contribute to improve data quantity and quality and, thus, allow developing more reliable models. A crucial role is foreseen for modelling tools as they can incorporate the cumulative impacts of multiple stressors, including the individual-level effects of plastics, to the population- and ecosystem level.

Abundance and composition of floating marine macro litter on the eastern sector of the Mediterranean Sea.

Floating marine litter is known to be currently a widespread pollution problem, especially in the Mediterranean basin, but records on the levels of floating debris in the eastern part are less known. By using an observation platform (32 days, 137 transects, 1784 km), this study provides evidence of a high amount of floating debris in the Levantine sub-basin, within the eastern Mediterranean Sea. The density of floating litter varied between 18 and 1593 items km-2 (average 232 ±â€¯325 items km-2), and small plastic debris accounted for >90% of the items surveyed. These values tend to be higher than densities reported for the central and western Mediterranean areas, which may be related to the circulation patterns and inputs from coastal sources. Significant correlations of floating macro litter density with wind force and sea state were found, thus strengthening the need to standardise protocols (including oceanographic variables) of visual surveys of floating debris.

Development of physical modelling tools in support of risk scenarios: A new framework focused on deep-sea mining.

Deep-sea mining has gained international interest to provide materials for the worldwide industry. European oceans and, particularly, the Portuguese Exclusive Economic Zone present a recognized number of areas with polymetallic sulphides rich in metals used in high technology developments. A large part of these resources are in the vicinity of sensitive ecosystems, where the mineral extraction can potentially damage deep-ocean life services. In this context, technological research must be intensified, towards the implementation of environmental friendly solutions that mitigate the associated impacts. To reproduce deep-sea dynamics and evaluate the effects of the mining activities, reliable numerical modelling tools should be developed. The present work highlights the usefulness of a new framework for risk and impact assessment based on oceanographic numerical models to support the adoption of good management practices for deep-sea sustainable exploitation. This tool integrates the oceanic circulation model ROMS-Agrif with the semi-Lagrangian model ICHTHYOP, allowing the representation of deep-sea dynamics and particles trajectories considering the sediments physical properties. Numerical simulations for the North Mid-Atlantic Ridge region, revealed the ability of ROMS-Agrif to simulate real deep-sea dynamics through validation with in situ data. Results showed a strong diversity in the particle residence time, with a dependency on their density and size but also on local ocean conditions and bottom topography. The highest distances are obtained for the smaller and less dense particles, although they tend to be confined by bathymetric constrains and deposited in deepest regions. This work highlights the potential of this modelling tool to forecast laden plume trajectories, allowing the definition of risk assessment scenarios for deep-sea mining activities and the implementation of sustainable exploitation plans. Furthermore, the coupling of this numerical solution with models of biota inhabiting deep-sea vent fields into ecosystem models is discussed and outlined as cost-effective tools for the management of these remote ecosystems.

Assessing the effects of temperature and salinity oscillations on a key mesopredator fish from European coastal systems.

A population dynamics model was developed to assess the short and long-term effects of temperature and salinity variations in the common goby Pomatoschistus microps in a Portuguese estuary (Minho estuary, NW Portugal). The population was divided into juveniles, females and males, which constituted the model's state variables. Linear regressions between the observed and the predicted density of juveniles, females and the total population were significant. Parameter's sensitivity and uncertainty analysis were estimated. The model was able to satisfactory describe the P. microps population dynamics, and thus was used to simulate the effects of climatic changes on the fish population. Simulations indicated that the common goby population is sensitive to both temperature and salinity changes. Overall, scenarios of more than 3 °C increase caused significant population decreases. Similarly, increased salinities led to a population shrinkage, whereas scenarios of salinity decrease generated an opposite variation on the population. According to the IPCC predictions for climatic tendencies, the population of the common goby will tend to decrease in the near future, experiencing marked oscillations (decrease or increase) during climatic extremes, namely droughts and floods, respectively. These results may be a useful for future planning and management of estuarine systems given that the common goby is an important species of estuarine food webs in many temperate ecosystems.

Assessing estuarine quality: A cost-effective in situ assay with amphipods.

In situ assays based on feeding depression can be powerful ecotoxicological tools that can link physiological organism-level responses to population and/or community-level effects. Amphipods are traditional target species for toxicity tests due to their high sensitivity to contaminants, availability in the field and ease of handling. However, cost-effective in situ assays based on feeding depression are not yet available for amphipods that inhabit estuarine ecosystems. The aim of this work was to assess a short-term in situ assay based on postexposure feeding rates on easily quantifiable food items with an estuarine amphipod. Experiments were carried out under laboratory conditions using juvenile Echinogammarus marinus as the target individual. When 60 Artemia franciscana nauplii (as prey) were provided per individual for a period of 30 min in dark conditions, feeding rates could be easily quantified. As an endpoint, postexposure feeding inhibition in E. marinus was more sensitive to cadmium contamination than mortality. Assay calibration under field conditions demonstrated the relevance of sediment particle size in explaining individual feeding rates in uncontaminated water bodies. An evaluation of the 48-h in situ bioassay based on postexposure feeding rates indicated that it is able to discriminate between unpolluted and polluted estuarine sites. Using the harmonized protocol described here, the in situ postexposure feeding assay with E. marinus was found to be a potentially useful, cost-effective tool for assessing estuarine sediment and water quality.

New isolates of Trichoderma antagonistic to Sclerotinia sclerotiorum / Novos isolados de Trichoderma antagônicos a Sclerotinia sclerotiorum

Abstract Forty-nine isolates of Trichoderma from the Brazilian Midwest were evaluated for their antagonistic activity in vitro against Sclerotinia sclerotiorum (causal agent of white mold), which were then identified based on their nuclear ribosomal ITS sequences. Paired culture tests showed that all isolates exhibited some antagonism, with a maximum of 77% mycelial inhibition and complete inhibition of sclerotia production. Two isolates were found to be the most promising biocontrol agents, considering both antagonistic parameters (CEN1253 - T. koningiopsis and CEN1265 - T. brevicompactum). Five different species were identified: T. harzianum (23), T. spirale (9), T. koningiopsis (8), T. brevicompactum (7) and T. asperellum (2). These isolates are stored in the Embrapa Fungi Collection for Biological Control and the information obtained in the experiments will be incorporated into the database of biological assets within the genetic resources information system (Allele) and be made available for further studies.

Resumo Quarenta e nove isolados de Trichoderma obtidos no centro-oeste do Brasil foram avaliados quanto a sua atividade antagônica in vitro contra Sclerotinia sclerotiorum (agente causal do mofo branco) e identificados com base nas sequências ITS do DNA ribossômico nuclear. Os testes de cultivo pareado mostram que todos os isolados exibiram algum antagonismo, com um máximo de 77% de inibiação micelial e inibição total da produção de escleródios. Dois isolados se destacaram como os mais promissores, considerando ambos os parâmetros avaliados (CEN1253 - T. koningiopsis e CEN1265 - T. brevicompactum). Cinco espécies diferentes foram identificadas: T. harzianum (23), T. spirale (9), T. koningiopsis (8), T. brevicompactum (7) and T. asperellum (2). Estes isolados estão armazenados na Coleção de Fungos para Controle Biológico da Embrapa e as informações obtidas nos experimentos serão incorporadas na base de dados de ativos biológicos, no sistema de informações de recursos genéticos, e disponibilizados para estudos futuros.

Ecological modelling and toxicity data coupled to assess population recovery of marine amphipod Gammarus locusta: Application to disturbance by chronic exposure to aniline.

A population agent-based model of marine amphipod Gammarus locusta was designed and implemented as a basis for ecological risk assessment of chemical pollutants impairing life-history traits at the individual level. We further used the model to assess the toxic effects of aniline (a priority hazardous and noxious substance, HNS) on amphipod populations using empirically-built dose-response functions derived from a chronic bioassay that we previously performed with this species. We observed a significant toxicant-induced mortality and adverse effects in reproductive performance (reduction of newborn production) in G. locusta at the individual level. Coupling the population model with the toxicological data from the chronic bioassay allowed the projection of the ecological costs associated with exposure to aniline that might occur in wild populations. Model simulations with different scenarios indicated that even low level prolonged exposure to the HNS aniline can have significant long-term impacts on G. locusta population abundance, until the impacted population returns to undisturbed levels. This approach may be a useful complement in ecotoxicological studies of chemical pollution to transfer individual-collected data to ecological-relevant levels.

Hypocholesterolaemic pharmaceutical simvastatin disrupts reproduction and population growth of the amphipod Gammarus locusta at the ng/L range.

Simvastatin (SIM), a hypocholesterolaemic drug, is among the most widely used pharmaceuticals worldwide and is therefore of emerging environmental concern. Despite the ubiquitous nature of SIM in the aquatic ecosystems, significant uncertainties exist about sublethal effects of the drug in aquatic organisms. Therefore, here we aimed at investigating a multi-level biological response in the model amphipod Gammarus locusta, following chronic exposures to low levels of SIM (64 ng/L to 8 µg/L). The work integrated a battery of key endpoints at individual-level (survival, growth and reproduction) with histopathological biomarkers in hepatopancreas and gonads. Additionally, an individual-based population modelling was used to project the ecological costs associated with long-term exposure to SIM at the population level. SIM severely impacted growth, reproduction and gonad maturation of G. locusta, concomitantly to changes at the histological level. Among all analysed endpoints, reproduction was particularly sensitive to SIM with significant impact at 320 ng/L. These findings have important implications for environmental risk assessment and disclose new concerns about the effects of SIM in aquatic ecosystems.

Predicting the variation in Echinogammarus marinus at its southernmost limits under global warming scenarios: can the sex-ratio make a difference?

Understanding the environmental parameters that constrain the distribution of a species at its latitudinal extremes is critical for predicting how ecosystems react to climate change. Our first aim was to predict the variation in the amphipod populations of Echinogammarus marinus from the southernmost limit of its distribution under global warming scenarios. Our second aim was to test whether sex-ratio fluctuations - a mechanism frequently displayed by amphipods - respond to the variations in populations under altered climate conditions. To achieve these aims, scenarios were run with a validated model of E. marinus populations. Simulations were divided into: phase I - simulation of the effect of climate change on amphipod populations, and phase II - simulation of the effect of climate change on populations with male and female proportions. In both phases, temperature (T), salinity (S) and temperature and salinity (T-S) were tested. Results showed that E. marinus populations are highly sensitive to increases in temperature (>2 °C), which has adverse effects on amphipod recruitment and growth. Results from the climate change scenarios coupled with the sex-ratio fluctuations depended largely on the degree of female bias within population. Temperature increase of 2 °C had less impact on female-biased populations, particularly when conjugated with increases in salinity. Male-biased populations were highly sensitive to any variation in temperature and/or salinity; these populations exhibited a long-term decline in density. Simulations in which temperature increased more than 4 °C led to a continuous decline in the E. marinus population. According to this work, E. marinus populations at their southernmost limit are vulnerable to global warming. We anticipate that in Europe, temperature increases of 2 °C will incite a withdrawal of the population of 5°N from the amphipod species located at southernmost geographical borders. This effect is discussed in relation to the distribution of E. marinus along the Atlantic coast.

Influence of some parameters on the germination assessment of mycopesticides.

The substantial negative impact of some parameters on the germination of low-quality conidia (high proportion of slow-germinating propagules) was demonstrated, whereas for high-quality batches their effect was small or even absent. Germination was increased as the initial hydration status of conidia immediately prior to suspension preparation was increased, being ca. 33% and 80% for dehydrated Metarhizium anisopliae propagules (water activity ≤0.314) from low- or high-quality batches after an 18 h incubation period, respectively, and 63% and 95% for hydrated propagules (water activity = 0.933). Germination of low-quality propagules also increased as the time dry conidia were kept in aqueous suspension prior to inoculation onto culture media (15 min, 3 or 24 h) or the incubation time at 25°C before counts (18, 48 or 72 h) was increased. Depending on treatment conditions, average germination of low-quality conidia varied from 53% to 98%. On the other hand, germination for high-quality conidia was always ≥94%. Regarding the relative humidity (RH) of the incubation atmosphere, the average germination rates for low-quality conidia on Potato Dextrose Agar (PDA) in Petri plates was 49%, while germination of these conidia on PDA blocks kept under lower RH inside plastic boxes was ≤23%. Use of lactophenol-staining and/or use of coverslips had a negative effect when germination assessment was performed for low-quality conidia, resulting in distorted counts or increased standard deviations compared to high-quality conidial batches. The occurrence of dislodged conidia (ungerminated conidia outside the inoculation zone due to hydraulic pressure exercised by addition of stains and/or coverslips added to the substrate by the time germination is assessed) was common place, whereas dislodged conidia were not seen in treatments with high-quality batches. This work underscores the importance of a number of parameters that anyone working with low-quality fungi needs to be cognizant of in their research.

Potencial antagônico de Trichoderma spp. originários de diferentes agroecossistemas contra Sclerotinia sclerotiorum e Fusarium solani / Antagonist potential of Trichoderma spp. from distinct agricultural ecosystems against Sclerotinia sclerotiorum and Fusarium solani

A partir de 40 amostras de solo, provenientes dos estados do Rio Grande do Sul, Minas Gerais, São Paulo, Bahia, Goiás e Tocantins, uma coleção de 230 isolados monospóricos de Trichoderma spp. foi estabelecida, usando o meio seletivo TSM. Com o objetivo de selecionarem-se isolados com potencial para controle biológico de doenças, foram conduzidos testes de pareamento de culturas em meio BDA, a 20 ºC para Sclerotinia sclerotiorum e a 25 ºC para Fusarium solani f.sp. phaseoli. Antagonismo contra os dois patógenos foi observado em 10 por cento dos isolados. Avaliações ao microscópio eletrônico de varredura de sete isolados selecionados in vitro mostraram que nem todos promoveram o hiperparasitismo dos patógenos, sugerindo a existência de outros mecanismos de antagonismo, como antibiose ou competição.

From 40 soil samples collected in the Rio Grande do Sul, Minas Gerais, São Paulo, Bahia, Goiás and Tocantins states, Brazil, a collection of 230 monosporic isolates of Trichoderma spp. was established using TSM selective media. In order to select efficient isolates for biological control, dual culture tests were carried out on PDA media at 20 ºC for Sclerotinia sclerotiorum and at 25 ºC for Fusarium solani f.sp. phaseoli. From the whole collection, 50 isolates presented antagonism against F. solani and 111 isolates to S. sclerotiorum. The antagonism against both pathogens was found in only 10 percent of the isolates. Scanning electron microscopy assessments with seven in vitro selected isolates showed that not all promoted hyperparasitism on the pathogens, suggesting the existence of other mechanisms of antagonism, as antibiosis or competition.

Intersexuality incidence, sex ratio fluctuations and intersex reproductive output as factors affecting the temporal variation of intersexed populations of the marine amphipod Echinogammarus marinus.

The occurrence of intersexuality in populations of the marine amphipod Echinogammarus marinus has been related to several causes, including anthropogenic contamination, microsporidian parasites and environmental sex determination (ESD). At the individual-level, the costs of intersexuality in E. marinus are reflected in reduced reproductive output of intersexes compared to normal specimens. The impacts of intersexuality and other reproductive disorders in nature will only be fully understood when individual-level effects are upscale to the population-level, which can be achieved through ecological models. The aim of the present work is to simulate the density variation of real populations of E. marinus with different intersexuality incidence, proportions of phenotypes and intersex female reproductive rates. To achieve that, we have incorporated empirical data obtained at the individual-level from three intersexed populations of E. marinus into a population dynamic model. The original model was calibrated and validated for a long-term stable population without intersexuality and sex ratio approximately 1. In the present version, the model includes an extra population group, intersex females, but the total sum of flow rates from juveniles to adults was kept the same as in the original model to respect the previous validation. In these circumstances, results indicate that the male-biased population is very sensitive to intersexuality, even at low incidence levels, and exhibits significant density decrease if conditions do not change. On the contrary, the two populations with a preponderance of female-biased sex ratios, low or moderate intersexuality incidence (up to an average value of 9% of the population) and with intersex reproductive rates corresponding to 80% the value of normal specimens, exhibit long-term persistence. The model is highly sensitive to adult female parameters (e.g. mortality), indicating that small variations on its values can significantly change population responses. Overall the present results suggest that, in spite the costs of intersexuality in E. marinus, the temporal variation of intersexed populations depend on intersex incidence level, sex ratio fluctuations and reproductive output of intersexes. Simulations also suggest that other processes (e.g. mortality rates of normal females) may play important roles in the maintenance of populations with intersexuality. In the future, it will be crucial to improve and validate this type of models with long-term observations and monitoring of intersexed populations.