Can we assess the ecological status of estuaries based on larval

fish

assemblages?

Régis Vinícius Souza Santosa,⁎, Sandra Ramosb,c, Ana Cristina Teixeira Boneckera

a Universidade Federal do Rio de Janeiro, Instituto de Biologia, Departamento de Zoologia, Laboratório Integrado de Zooplâncton e
Ictioplâncton, Av. Carlos Chagas Filho,

373, Prédio do CCS, Bloco A, 21.941-902 Rio de Janeiro, RJ, Brazil

b CIIMAR - Interdisciplinary Centre of Marine and Environmental Research, ECOBIOTEC, Avenida General Norton de Matos, S/N,
4450-208 Matosinhos, Portugal

c Institute of Estuarine and Coastal Studies, University of Hull, Hull HU6 7RX, UK

ABSTR ACT

Fish larvae of four SE Brazilian estuaries were investigated to assess if the larval fish assemblages reflect the

ecological status of estuaries. All samples were collected in the same water mass to guarantee similar natural

water parameters, assuring that major differences among estuaries were related to anthropogenic pressures.

Water temperature, oxygen, pH, chlorophyll a, faecal coliforms, nutrient load and total particulate matter were

obtained at each sampling area. A pressure index was used to assess the overall anthropogenic pressures acting in

each estuary. Results showed that fish larvae were sensitive to water contamination, reducing the diversity and

especially exhibiting a high dominance of few species. Furthermore, this study reinforced the idea that the high

sensitivity of fish larvae can increase the accuracy of the environmental assessments when tackling short-time

events of hydrological controls (physical barriers and control of the freshwater input), representing an advance

in the water ecological quality assessments.

1. Introduction

Estuaries are essential fish habitats, compiling several functions

such as nursery grounds, migration routes and refuge areas for a variety

of fish species (Cattrijsse and Hampel, 2006). Nevertheless, numerous

anthropogenic perturbations affect estuarine environments (Elliott and

Whitfield, 2011), contributing to habitat alteration and changes in the

structure and dynamics of biotic communities (Kennish, 2002), compromising

the long-term ecological function of these ecotones (e.g.

Whitfield and Elliott, 2002; Gilliers et al., 2006; Coates et al., 2007; Le

Pape et al., 2007). In response to the increasing degradation of marine

ecosystems observed worldwide (Halpern et al., 2007, 2008), several

studies have assessed the ecological status of aquatic ecosystems and

the impacts of anthropogenic activities (e.g. Borja et al., 2013; Ruaro

and Gubiani, 2013; Martinez-Haro et al., 2015), in order to trace effective

management plans and mitigation actions in these ecosystems.

In estuarine environments, several groups of organisms have been

used as quality indicators of ecosystem health, mainly macroinvertebrates

and fishes (e.g. Harrison et al., 2000; USEPA, 2000;

Alvarez et al., 2013; Nebra et al., 2014; Alves et al., 2015). Ecological

indicators based on fish communities have been recognized as useful

tools to assess anthropogenic impacts affecting estuarine fish

communities (Whitfield and Elliott, 2002; Harrison and Whitfield,

2006; Borja and Dauer, 2008; Fonseca et al., 2013). Typically, a fish

community facing increased anthropogenic induced stress tends to

show a decreased of number and abundance of species with high habitat

and environmental specificities, i.e. sensitive and low tolerance

fish species, while generalist species tend to increased dominance, resulting

in a simplified community structure (e.g. Odum, 1983; Karr and

Chu, 1999; Fonseca et al., 2013). Nevertheless, the use of fishes as indicators

of environmental quality can also have limitations because of

their high mobility, temporal variability (seasonality) and specific

sampling requirements (Whitfield and Elliott, 2002; Harrison and

Whitfield, 2004).

Given these limitations, early larval stages of fishes can be considered

good indicators of ecosystem integrity, since they have reduced

sampling costs and sampling bias, low mobility (planktonic phase) and
sensitivity to reflect anthropogenic pressures acting on a water body

(Ramos et al., 2015). Indeed, fish larvae show a high vulnerability to

environmental changes (Ramos et al., 2006a; Eick and Thiel, 2014;

Strydom, 2015; Santos et al., 2017), highlighting their importance as

pollution indicators (Longwell et al., 1992; Gordina et al., 2001;

Westernhagen et al., 2001). Ramos et al. (2012) demonstrated that

estuaries with different levels of anthropogenic pressures exhibit

distinct larval fish assemblages, and also emphasized the potential role

of larval stages of fishes as indicators of hydrological pressures that are

difficult to measure (Ramos et al., 2015). Moreover, the use of fish

larvae to assess ecological conditions of estuaries integrates valuable

information about spawning and nurseries areas, and about connectivity

pathways between marine/riverine habitats.

Estuarine fish larvae assemblages continually vary in time and

space, according to reproductive seasons of the species and due to environmental

fluctuations (Barletta-Bergan et al., 2002; Ramos et al.,

2006a; Montoya-Maya and Strydom, 2009; Strydom, 2015). Consequently,

the environmental conditions may interfere with ecological

quality assessment when fish larvae are used as quality indicators, as

well as temporal variability patterns associated with natural fluctuations

of species-specific reproductive seasons, considered as a major

driver of the estuarine larval fish assemblages (Rakocinski et al., 1996).

The Estuarine Quality Paradox (Elliott and Quintino, 2007), which

suggests that anthropogenic stress is difficult to detect in estuaries because

their existing communities also show the natural variability, has

been reflected in many studies of ecological quality assessments (e.g.

Courrat et al., 2009; Dauvin and Ruellet, 2009; Ramos et al., 2015;

Tweedley et al., 2015). This is a particular problem in estuaries since

their natural environmental conditions vary both spatially and temporally

(Sheaves, 2016). Thus, the constancy of environmental characteristics

is fundamental to the spatial comparisons of the category and

level of anthropogenic pressures, by removing the potentially confounding

effects of natural variability on fish assemblages (Elliott and

Quintino, 2007). In this context, we tested the hypothesis that estuaries

with similar environmental characteristics, but distinct anthropogenic

pressures, carry distinctly different larval fish assemblages, by investigating

the larval fish assemblages of four similar tropical estuaries

with different types and levels of human pressures.

2. Material and methods

2.1. Methodological approach

To test the hypothesis that estuaries with similar environmental

characteristics suffering from specific anthropogenic pressures comprise

distinct larval fish assemblages, we selected four estuaries located

in the same biogeographic area of SE Brazil, in order to remove assemblage

differences due to geographic location of the estuaries. In

addition, all samples were collected in the same water mass (salinity

15–25) to guarantee similar natural water parameters allowing interestuarine

comparisons. Considering that estuarine larval fish assemblages

are highly seasonal (Ramos et al., 2006b; Arévalo-Frías and

Mendoza-Carranza, 2015; Strydom, 2015), we analyzed rainy/dry

seasons separately, ensuring that possible assemblages differences were

not related to seasonal influences. Hence, we restricted inter-estuarine

variability in terms of natural environmental conditions, assuring that

major differences among estuaries were related to type and level of

anthropogenic pressures. The anthropogenic pressures acting in the

estuaries were assessed based on contamination indicators, as nutrient

load (nitrate, nitrite, ammonia and phosphate) and sewage contamination

determined by microbial faecal coliforms contamination.

Also, a pressure index was calculated for each estuary based on human

activities and hydro-morphological pressures.

2.2. Estuarine models

Four estuaries of SE Brazil, Macaé, São João, Bracuí and Perequê-

Açu were investigated (Fig. 1). Macaé (22° 22′ S and 41° 46′ W) and São

João (22° 35′ S and 41° 59′ W) are larger systems directly connected to

the ocean with 130 km and 120 km in length, respectively; and total

area of 1765 km2 and 2160 km2, respectively. Bracuí (22° 57′ S and 44°

23′ W) and Perequê-Açu (23° 13′ S and 44° 42′ W) are smaller systems

with 32 km in length and a total area of 190 km2, and 22 km and area of

110 km2, respectively; connected to a coastal bay (Ilha Grande Bay).

The four estuaries are shallow (mean depth of 2–4 m), channel type,

microtidal and partially mixed, with semidiurnal regime.

The levels of anthropogenic pressures vary among the estuaries. In

Macaé and Perequê-Açu estuaries, the unplanned urbanization process

promoted the removal of mangroves in the estuarine region and discharge

large loads of organic matter and other compounds of human

waste without treatment directly into the water (INEA, 2014, 2015).

Both estuaries are used for navigation, mainly tour boats in Perequê-

Açu Estuary, which relies on constant dredging to improve navigability.

Macaé Estuary is used as navigation route for support vessels to oil and

gas drilling platforms. The freshwater input of Macaé River is highly

controlled by water abstraction for human consumption, industrial use

and to cool the power-producing equipment in thermoelectric plants

(INEA, 2014). Additionally, there are several dams, agricultural activities

and commercial sand extraction along the Macaé River. São João

Estuary still holds mangrove areas in the middle and lower sections,

although there are agricultural and fishing activities that might represent

a potential pressure. Also, the main river is obstructed by a

dam, which controls the river flow reaching the São João Estuary.

Bracuí Estuary still has natural banks with mangrove areas without

hydro-morphological pressures, and human activities are limited to

some aquaculture and agricultural activities.

2.3. Sampling and sample processing

Sampling surveys were conducted every two months between May

2013 and March 2015. All samples were taken in the same salinity

range (15–25) during nightly ebb tides. Triplicates of surface trawls

were performed in each estuary with a Bongo net (500 μm) and the

volume of the water filtered was determined by a flowmeter (General

Oceanics®). At each sampling area, subsurface (0.5–1 m depth) water

temperature and pH were obtained by a multiparameter probe (Hach

HQ40d) and water samples were collected for further analyses.

Chlorophyll a and inorganic nutrients (nitrate, nitrite, ammonia and

phosphate) were determined following previously described methods

(Grasshoff et al., 1999). Oxygen saturation was calculated as the percentage

of dissolved oxygen concentration relative to the temperature

and salinity of the water, and the dissolved oxygen concentration was

determined by the Winkler method (Grasshoff et al., 1999). Total particulate

matter was determined following APHA (2005) method. Faecal

coliforms were determined by the fermentation method in multiple

tubes, which determines the Most Probable Number (MPN) of faecal

coliforms per 100 mL water sample (APHA, 2005). Monthly precipitation

data were obtained from the National Agency of Water (ANA,

2015). Fish larvae were sorted from 144 ichthyoplankton samples and

identified to the highest possible taxonomic separation.

2.4. Data analysis

Months were aggregated into two seasonal groups, namely: dry

comprising May, July, September and November; and rainy comprising

January and March. All analyses were performed for each season separately.

Differences in water parameters (temperature, pH, total particulate

matter, chlorophyll a and oxygen saturation) and contamination

indicators (nutrients and faecal coliforms) among the four estuaries

were analyzed by one-way MANOVA (Multivariate Analysis of

Variance) with estuary as fixed factor (Sokal and Rohlf, 1995; Zar,

1996). One-way ANOVA was used to investigate differences in the

community descriptors (abundance, number of species and guilds, diversity

and equitability) among estuaries (Zar, 1996). All variables,

excepting pH, were log transformed in order to stabilize the variance

and to fit data to a normal distribution, fulfilling one of the ANOVA and

MANOVA assumptions. Homogeneity of variance was tested by a Cochran

test, and whenever variance was still heterogeneous, conclusions

from ANOVA and MANOVA results were only accepted for those cases

where significance levels were< 0.01. Furthermore, in the event of

significance, a posteriori LSD (Least Significant Difference) test was

used to determine which means were significantly different at a 0.05

level of significance.

A Pressure Index (PI), adapted from Aubry and Elliott (2006), assessed

anthropogenic pressures acting in each estuary. PI included 13

descriptors relevant for larval stages of fishes, namely: habitat loss,

dredging, percentage of land for industrial and agricultural use, population

density, presence of marinas and ports, tourism and recreation,

physical barriers (dams), aquaculture, fisheries, freshwater input

(amount of freshwater that naturally reaches the estuary) and sewage

pollution (discharge of loads of organic matter and other compounds of

human waste without treatment). These descriptors may have apparently

included an element of double counting or redundancy, which can

lead to the over-weighting of particular pressures. However, we consider

that a single number of indicators surely cannot capture the

complexity involved in environmental state (Purvis and Hector, 2000;

Derous et al., 2007; Borja et al., 2014), and that the ecosystem-based

approach implies that overall environmental status can only be appropriately

described by combining the different aspects of the environment

into a holistic view (Berg et al., 2015). The 13 selected descriptors

were classified as: no change (0), very low (1), low (2),

moderate (3), high (4) and very high change (5), and the PI resulted

from the average value of the individual pressures scores. Simple

average of all descriptors is not necessarily the best solution in every

circumstance, considering that different descriptors meet various

screening criteria differently (Shin et al., 2010; Borja et al., 2014,

2016). However, an adequate basis for weighting descriptors is not

always available and in such cases equal weighting is recommended by

Ojaveer and Eero (2011).

Differences on the larval fish assemblages were investigated at the

structural (abundance, diversity and equitability) and functional (ecological

guilds) levels. Abundance data were standardized into number

of larvae per 100 m3 of water filtered. Diversity of the larval fish assemblage

was expressed by Shannon-Wiener index (H′) based on natural

log (Shannon and Weaver, 1963), and equitability was measured

by Pielou's evenness index (J′) (Pielou, 1966). Fish species were assigned

to an ecological guild according to their estuarine use patterns,

following Elliott et al. (2007): amphidromous species (AM), anadromous

species (AN), catadromous species (C), estuarines (ES), freshwater

species (F), marine migrants (MM; spawn at sea and regularly enter

estuaries in large numbers, including marine species using estuaries as

nursery grounds) and marine stragglers (MS; spawn at sea and enter

estuaries accidentally in low numbers).

Inter-estuarine differences in terms of larval fish assemblage were

investigated by one-way analysis of similarity (ANOSIM) (Clarke and

Warwick, 1994), at a significance level of P < 0.05 and considering R

statistic > 0.5 (Clarke and Warwick, 1994). The tests were based on a

Bray-Curtis rank similarity matrix, calculated using log transformed

[log 10(x +1)] fish larval data. Non-metric multidimensional scaling

(nMDS) was used to investigate similarities among estuarine samples,

based on the previously described Bray–Curtis similarity matrix (Bray

and Curtis, 1957). Similarity percentages (SIMPER) (Clarke, 1993) were

used to identify the percentage contribution of each taxon to the

average dissimilarity between samples of the various estuaries pair

combinations. Multivariate analyses were performed with the software

package PRIMER 6 (Plymouth Routines Multivariate Ecological Research)

(Clarke and Warwick, 1994).

To evaluate the influence of environmental variables on the larval

fish assemblages of each estuary a canonical correspondence analysis

(CCA) (Ter Braak, 1986) was performed using CANOCO software

(version 4.5, Microcomputer Power, Ithaca, NY). Monthly precipitation,

water temperature, oxygen saturation, pH, TPM, chlorophyll a, nitrate,

nitrite, ammonia, phosphate, faecal coliforms and PI were standardized

and used as explanatory environmental variables. All 59 taxa were used

in the CCA in order to have a comprehensive picture of larval fish

distributions. Larval abundances were square root transformed to normalize

the data and downweighting of rare species was selected. Statistical

significance of the correlations between explanatory and species

variables extracted from the CCA was determined by the Monte–Carlo

test, based on 499 permutations. Environmental variables were considered

to be biologically important when inter-set correlation coefficients

were |≥0.4| (Hair et al., 1984; Rakocinski et al., 1996).

3. Results

3.1. Environmental characteristics

In general, high precipitation values were observed during the rainy

season in the four estuaries (Table 1), although mean seasonal precipitation

varied among estuaries. During the rainy period, Bracuí and

Perequê-Açu estuaries registered the highest precipitation values

(Table 1). Macaé Estuary registered the lowest mean precipitation

during both rainy and dry seasons. MANOVA results showed some

significant differences in particular water parameters, namely temperature

during the dry season and TPM, oxygen and chlorophyll a

during the rainy season (Table 1). During the dry season, Perequê-Açu

Estuary reached the highest water temperature (Wilks' lambda = 0.40,

P < 0.05) and Macaé Estuary registered significantly lower dissolved

oxygen (Wilks' lambda = 0.40, P < 0.05). During the rainy season,

Macaé Estuary exhibited significantly lower water temperature (Wilks'

lambda = 0.08, P < 0.05) and highest chlorophyll a concentration

(Wilks' lambda = 0.08, P < 0.05). In addition, Macaé Estuary presented

significantly high TPM values (Wilks' lambda = 0.08,

P < 0.05) during both seasons. The remaining water characteristics

did not vary among estuaries.

3.2. Contamination indicators

Contamination indicators differed significantly among the four estuaries

during both seasons (dry: Wilks' lambda = 0.25, P < 0.001;

rainy: Wilks' lambda =0.05, P < 0.01), with Macaé and Perequê-Açu

estuaries exhibiting lower water quality than Bracuí and São João estuaries.

Overall, Macaé and Perequê-Açu estuaries were more contaminated

by faecal coliforms and carried a higher nutrient load, particularly

higher concentrations of nitrate, ammonia and phosphate

(Table 2).

3.3. Pressure index

According to the pressure index (PI) the four estuaries differed in

terms of anthropogenic pressures, with Macaé Estuary classified as

highly impacted and the most impacted estuary, followed by Perequê-

Açu Estuary, classified as medium impacted estuary (Table 3). São João

and Bracuí estuaries were the least impacted systems, classified as low

and very low impacted, respectively (Table 3). Macaé Estuary suffers

from several human pressures, mainly alteration of the amount of

freshwater that naturally reaches the estuary, large-scale habitat loss, a

high percentage of land use for agriculture, sewage pollution and

dredging activities. In Perequê-Açu Estuary, habitat loss, tourism and

recreation, sewage pollution and dredging were the most severe pressures

observed. The most important pressures detected in São João

Estuary were the percentage of land for agricultural use and the

freshwater input, which is controlled by a large dam located upstream.

Bracuí Estuary exhibited few pressures, associated with some aquaculture

and land use for agriculture.

3.4. Larval fish assemblages

The larval fish assemblages varied significantly among the four estuaries

during both seasons, not only in terms of abundance (dry:

F= 13.07, P < 0.001; rainy: F= 8.58, P < 0.001), but also the

number of species (dry: F= 18.63, P < 0.001; rainy: F =2.86,

P =0.05), diversity (dry: F= 23.74, P < 0.001; rainy: F= 17.08,

P < 0.001) and equitability (dry: F= 15.36, P < 0.001; rainy:

F= 17.19, P < 0.001). Perequê-Açu Estuary showed significantly

higher abundances (P < 0.001) during both seasons, with a mean of

291 ± 306 larvae 100 m−3 during the dry season, and 2791 ± 3901

larvae 100 m−3 in the rainy period (Table 4). Such high abundances

were associated with significantly (P < 0.001) lower diversity (dry:

H′ =0.8 ± 0.4, rainy: H′= 0.5 ± 0.4) and equitability values (dry:

J′ =0.5 ± 0.3, rainy: J′= 0.3 ± 0.3). During the dry season, Macaé

larval fish assemblages were significantly (P < 0.01) more abundant

than those of São João and Bracuí, with an average of 111 ± 94 larvae

100 m−3 (Table 4), and comprised the significantly (P < 0.001)

highest number of species of all study (Table 4).

The nMDS grouped Macaé and Perequê-Açu samples on opposite

sides of the plots, while Bracuí and São João samples were dispersed in

both plots (Fig. 2A–B). According to ANOSIM results, the structure of

the larval fish assemblages differed among the four estuaries, during the

dry (R = 0.6, P < 0.01) and rainy seasons (R = 0.5, P < 0.01).

During both seasons, Perequê-Açu samples were highly dominated by

Anchoviella lepidentostole (Supplementary data), which was responsible

for at least 20% of the average dissimilarity between Perequê-Açu Estuary

and the other estuaries. During the dry period, Macaé assemblages

were characterized by high abundances of Micropogonias furnieri

that reached a mean of 40 larvae 100 m−3, while in the other estuaries

did not surpass 2 larvae 100 m−3 (Supplementary data). On the other

hand, São João and Bracuí samples were not clearly separated from

each other (R < 0.5, P < 0.01), presenting similar larval fish assemblages,

characterized by high abundances of Anchoviella spp. in

both seasons (Supplementary data).

The larval fish assemblages of the four estuaries studied comprised

seven functional guilds (AN, ES, MM, MS, AM, C, F) (Supplementary

data). The majority of the species collected in the four estuaries were

AN, ES, MM MS (Fig. 3). In dry surveys, AN was the main group in São

João, Bracuí and Perequê-Açu estuaries, which reaching 47%, 54% and

79% of the total larvae captured in each estuary, respectively (Fig. 3A).

On the other hand, Macaé larval fish assemblage was dominated by MM

that represented 53% of the total larvae (Fig. 3A). In rainy surveys, AN

was the most abundant group in the four estuaries, reaching 36%, 68%,

80% and 98% of the total catch in Macaé, São João, Bracuí and Perequê-

Açu estuaries, respectively (Fig. 3B). The number of ecological

guilds was significantly higher in Macaé Estuary during the dry

(F = 3.52, P < 0.05), that comprised two further ecological guilds

(AM and F), but did not differ during the rainy period (F = 1.30,

P = 0.28). According to ANOSIM results, the guild structure of the four

estuaries did not vary significantly during the dry season (R = 0.2,

P = 0.001) or the rainy season (R = 0.3, P= 0.001). Also, estuarine

samples of the four estuaries were not separated by the nMDS of the dry

(Fig. 4A) and rainy seasons (Fig. 4B).

In order to allow inter-season comparisons, CCA analyses of the dry

and rainy seasons were performed separately. During the dry season

(Fig. 5A), the first CCA axis (CCA1) was negatively correlated with
temperature and positively correlated with TPM, ammonia and PI

(Table 5). And the second CCA axis (CCA2) was negatively correlated

with nitrate and PI (Table 5). Samples tended to cluster according to

their estuarine origin, with the four estuaries discriminated along the

two CCA axes. Bracuí and Perequê-Açu samples clustered on the left

side of CCA1 associated with high water temperature and lower nutrient

load, and were also separated along CCA2, with Perequê-Açu

samples positively correlated with nitrate and PI (Fig. 5A). São João

and Macaé samples were positively correlated with TPM and ammonia

(clustering on the right side of CCA1) and were discriminated by CCA2,

since Macaé samples were positively correlated with nitrate and PI

(Fig. 5A).

CCA analysis of the rainy season (Fig. 5B) showed that the first CCA

axis (CCA1) was negatively correlated with temperature and oxygen,

while the second CCA axis (CCA2) was positively correlated with nutrient

load (nitrate, nitrite, ammonia and phosphate) and PI (Table 5).

Perequê-Açu samples were isolated on the left side of CCA1 associated

with high water temperature and dissolved oxygen (Fig. 5B). Bracuí,

São João and Macaé samples were negatively correlated with temperature

(clustering on the right side of CCA1) and were mainly discriminated

by CCA2, with Macaé samples positively correlated with

nutrient load and PI and separated from the less contaminated São João

and Bracuí samples (Fig. 5B).

4. Discussion

One of the most critical phases in fish life cycle is the larval stage

(Miller and Kendall, 2009), which is particularly sensitive to environmental

conditions (Ramos et al., 2006a; Eick and Thiel, 2014; Strydom,

2015), including hydrological alterations of estuarine environment

(Ramos et al., 2012, 2015) and contamination (Longwell et al., 1992;

Gordina et al., 2001; Westernhagen et al., 2001). The level of contamination

impact on the estuarine environments may vary according

to estuarine geomorphology and hydrodynamics, specifically the degree

of connectivity with the marine ecosystem. This characteristic directly

influences the water residence time and dictates the time over which

the fish are exposed to any stressors (Elliott and Whitfield, 2011).

However, in the present study, the descriptors associated with the larval

fish assemblages were affected more by the anthropogenic pressures on

the systems than by their degree of connectivity with the ocean (i.e. if

they flow directly into the open sea or into large coastal bays). Differences

in the structure and functioning of assemblages associated with

the estuarine impact level may be identified by the different patterns of

fish species occurrence and abundance in the four estuaries.

In Perequê-Açu Estuary, the reduced ecosystem health (high nutrient

load and faecal contamination) negatively influenced the larval

fish assemblage, which showed some environmental stress symptoms,

namely reduced number of species, low diversity and especially a high

dominance of few species (Odum, 1983; Fausch et al., 1990). Macaé

Estuary was highly contaminated by sewage, which might have potentially

increased respiration processes and consequently lead to an

oxygen decrease of water (Thurman, 1985). This scenario might have

been the reason for the lower oxygen detected in Macaé dry samples,

since high faecal coliforms coincided with high nutrient load and TPM

values. Ramos et al. (2012) observed a similar effect in a small urban

estuary where the high contamination levels of faecal indicators and

high concentrations of dissolved organic carbon were associated to an

oxygen decreased of the water. However, Macaé larval assemblage

structure did not reflect the effect of high sewage contamination and

nutrient load observed in this study. In this case, the local pressure level

probably was not sufficiently high to cause significant impacts in the

structure of larval fish assemblages. This ability of the estuary to absorb

an amount of a contaminant added to the system is regarded as the

“assimilative capacity”. It can vary according to the part of the estuary

that receives the inputs, as well as being related to environmental

factors, such as water circulation (McLusky and Elliott, 2004). According

to Elliott and Jonge (2002), some estuarine areas have high

assimilative capacity often caused by increased dilution or flushing.

Considering this, the large water abstraction and flow changes observed

in Macaé Estuary may have increased the seawater intrusion and,

consequently, the assimilative capacity of this estuary by dilution. On

the other hand, the seawater intrusion may also have favored the migration

of several marine species, which may have been the reason for

this estuary to be clearly more diverse, despite the high contamination

level. Recently, Teichert et al. (2016) demonstrated that the flow

changes are essential explaining components of fish distribution in estuaries

and, in addition with water pollution and oxygen depletion, are

stressors classified at the ranks of priority in the restoration of fish

ecological quality in estuaries.

It has been acknowledged that the pollution in estuaries has a relevant

impact on fish species, not only in terms of the community

structure (reduced species diversity), but also at the function (reduced

abundance of migrant species and disturbance sensitive species) (e.g.

Harrison and Whitfield, 2004; Franco et al., 2006; Rochette et al., 2010;

Ramos et al., 2012; Fonseca et al., 2013). In this study, the results

showed that all estuaries exhibited similar number of ecological guilds,

but in different relative proportions of occurrence. In fact, the anthropogenic

pressures appeared to influence in different ways the estuarine

use patterns by fishes. River flow changes caused by water abstraction

and presence of barriers affected the migration routes of fish species,

influencing mainly the occurrence of the marine migrant M. furnieri,

and anadromous species A. lepidentostole and Anchoviella spp. The

croaker M. furnieri is a marine migrant species that systematically uses

estuarine environment along its ontogeny (Vieira, 2006; Costa et al.,

2013). Spawning occurs in the coastal zone (Macchi and Christiansen,

1996; Acha et al., 1999; Simionato et al., 2008; Braverman et al., 2009)

and eggs and larvae enter in the estuary associated with the intrusion of

salt water (Sinque and Muelbert, 1997; Elliott and Hemingway, 2002).

In this way, the river discharges may regulate croaker estuarine recruitment

by promoting high recruitment during low discharge periods

(Acha et al., 2012). Following this, the higher abundances of M. furnieri

in Macaé Estuary appears to be linked to the altered hydrological regime

caused mainly by water abstraction, which favored the estuarine

migration from the coastal area through the seawater intrusion.

Anadromous species, such as the anchovies A. lepidentostole and

Anchoviella spp., may also respond directly to hydro-morphological

pressures caused by dams. These species are characterized by migrating

upstream reaching the rivers where they spawn (Paiva-Filho et al.,

1986). Thus, the absence of physical barriers interfering with fish migratory

routes, could have contributed to the higher abundances of A.

lepidentostole and Anchoviella spp. in Perequê-Açu and Bracuí estuaries.

Furthermore, the warmest water of Perequê-Açu Estuary might also

have contributed to impressive high densities of anchovies, since their

abundance, spawning and distribution have been already reported as

positively associated with temperature (Silva and Araújo, 2000; Ribeiro

et al., 2014; Santos et al., 2017). The high temperature in Perequê-Açu

Estuary during both seasons could have been a result of the influence of

a warmer and oligotrophic seawater mass as Tropical Water (TW),

which is predominant in this region as well as in the most of the Brazilian

continental margin (BCM) (Lopes et al., 2006). On the other

hand, this typical warming of Brazilian tropical marine ecosystems was

interestingly less pronounced in Macaé Estuary during the rainy, although

the seawater intrusion was high due to lower precipitation

observed in this system, associated with the large water abstraction.

The low water temperatures and also high chlorophyll a concentrations

observed in Macaé rainy samples could have been a result of the influence

of a colder and nutrient-rich seawater mass as South Atlantic

Central Water – SACW (Acha et al., 2004; Braga and Niencheski, 2006).

The SACW intrusion over the shelf is more frequent during the rainy

season (spring-summer), when prevailing northeast winds cause Ekman

transport of surface waters offshore (Rodrigues and Lorenzzetti, 2001;

Castro et al., 2006a, 2006b; Calado et al., 2010; Campos et al., 2013;

Macedo-Soares et al., 2014), and its effect in the biota and seawater

properties has been observed along the Brazilian coast, mainly between

Cape São Tomé (22°S) and Cape Frio (23°S) (e.g. Macedo-Soares et al.,

2014; Moser et al., 2014; Suzuki et al., 2015). This is other example of

how the catchment events can contribute to processes and changes in

the environmental conditions within the estuary.

Macaé, São João, Bracuí and Perequê-Açu estuaries carried distinctly

different larval fish assemblages as a reflection of different environmental

status. Macaé and Perequê-Açu estuaries emerged as more

impacted than Bracuí and São João estuaries. Faecal contamination and
nutrient load were the main anthropogenic factors influencing the

larval fish assemblages in Perequê-Açu Estuary, while in Macaé Estuary

the water contamination effect on community structure was apparently

diminished by seawater/marine species intrusion resulting from the

other anthropogenic factor: the hydrological manipulations. Finally,

this study showed the TW/SACW effect on the estuarine larval fish

assemblages and water properties, emphasizing the need to develop

more studies in order to more accurately define the limits of these

seawater masses influences in this region.

5. Conclusions

The present study showed that fish larvae were sensitive to water

contamination, reducing the diversity and especially exhibiting a high

dominance of few species. Also, hydro-morphological pressures caused

by physical barriers and control of the freshwater input interfered in

migratory fish routes. Thus, this study reinforced the idea that the high

sensitivity of fish larvae can increase the accuracy of the environmental

assessments, mainly when tackling short-time events of hydrological

manipulations or water pollution, representing an advance in the assessment

of water ecological quality assessments. Also, the present

study gives strategically important information for improved estuarine

management by integrating valuable information about the environmental

status of estuarine ecological functions, as nursery areas, and

the connectivity between marine and freshwater habitats, considered

vital to many fish species

Español

¿Podemos evaluar el estado ecológico de los estuarios basados en ensambles

de peces larvales?

Se investigaron larvas de peces de cuatro estuarios brasileños SE para

evaluar si las agrupaciones de peces larvales reflejan el estado ecológico
de los estuarios. Todas las muestras se recogieron en la misma masa de agua
para garantizar parámetros similares de agua natural, asegurando que las
diferencias principales entre estuarios estuvieran relacionadas con las
presiones antropogénicas. La temperatura del agua, el oxígeno, el pH, la
clorofila a, los coliformes fecales, la carga de nutrientes y la materia
particulada total se obtuvieron en cada área de muestreo. Se utilizó un índice
de presión para evaluar las presiones antropogénicas globales que actúan en
cada estuario. Los resultados mostraron que las larvas de peces eran
sensibles a la contaminación del agua,
especialmente exhibiendo un alto predominio de pocas especies. Además, este
estudio reforzó la idea de que la alta
La sensibilidad de las larvas de peces puede aumentar la precisión de las
evaluaciones ambientales cuando se
eventos de los controles hidrológicos (barreras físicas y control de la
entrada de agua dulce), lo que representa un avance
en las evaluaciones de calidad ecológica del agua
Introducción
Los estuarios son hábitats esenciales de los peces, compilando varias
funciones
tales como viveros, rutas de migración y áreas de refugio para una variedad
de especies de peces (Cattrijsse y Hampel, 2006). Sin embargo, numerosos
perturbaciones antropogénicas afectan los ambientes estuarinos (Elliott y
Whitfield, 2011), contribuyendo a la alteración del hábitat y cambios en la
estructura y dinámica de las comunidades bióticas (Kennish, 2002),
comprometiendo
la función ecológica a largo plazo de estos ecótonos (p.
Whitfield y Elliott, 2002; Gilliers et al., 2006; Coates et al., 2007; Le
Pape et al., 2007). En respuesta a la creciente degradación de los recursos
observados en todo el mundo (Halpern et al., 2007, 2008), varios
estudios han evaluado el estado ecológico de los ecosistemas acuáticos y
los impactos de las actividades antropogénicas (por ejemplo, Borja et al.,
2013, Ruaro
y Gubiani, 2013; Martinez-Haro et al., 2015), con el fin de trazar planes
de manejo efectivos y acciones de mitigación en estos ecosistemas.
En ambientes estuarinos, se han utilizado varios grupos de organismos como
indicadores de calidad de la salud de los ecosistemas, principalmente
macroinvertebrados
y peces (por ejemplo, Harrison et al., 2000, USEPA, 2000;
Alvarez et al., 2013; Nebra et al., 2014; Alves et al., 2015). Ecológico
los indicadores basados en comunidades de peces han sido reconocidos como
instrumentos útiles para evaluar los impactos antropogénicos que afectan a
las comunidades de peces estuarinos (Whitfield y Elliott, 2002, Harrison y
Whitfield,
2006; Borja y Dauer, 2008; Fonseca et al., 2013). Típicamente, una comunidad
de peces que enfrenta un mayor estrés inducido por la actividad antropogénica
tiende a mostrar una disminución del número y la abundancia de especies con
altas especificidades de hábitat y medio ambiente, es decir, especies de
peces sensibles y de baja tolerancia, mientras que las especies generalistas
tienden a aumentar la dominancia, resultando
en una estructura comunitaria simplificada (por ejemplo, Odum, 1983; Karr
and
Chu, 1999; Fonseca et al., 2013). Sin embargo, el uso de peces como
indicadores
calidad ambiental también pueden tener limitaciones debido a
su alta movilidad, variabilidad temporal (estacionalidad) y
requisitos de muestreo (Whitfield y Elliott, 2002, Harrison y
Whitfield, 2004).
Dadas estas limitaciones, las etapas larvarias tempranas de los peces pueden
considerarse buenos indicadores de la integridad del ecosistema, ya que han
reducido
costos de muestreo y sesgo de muestreo, baja movilidad (fase planctónica)
y sensibilidad para reflejar las presiones antropogénicas que actúan sobre
un cuerpo de agua (Ramos et al., 2015). De hecho, las larvas de peces muestran
una alta vulnerabilidad a los cambios ambientales (Long et al., 2006a, Eick
y Thiel, 2014, Strydom, 2015, Santos y otros, 2017), destacando su
importancia como indicadores de contaminación , Gordina et al., 2001,
Westernhagen et al., 2001). Ramos et al. (2012) demostraron que los estuarios
con diferentes niveles de presiones antropogénicas

distintas asociaciones larvales de peces, y destacó también el

estadios larvales de los peces como indicadores de las presiones hidrológicas
difícil de medir (Ramos et al., 2015). Además, el uso de pescado
las larvas para evaluar las condiciones ecológicas de los estuarios,
información sobre zonas de desove y viveros, y sobre la conectividad
vías entre hábitats marinos / ribereños.
Los grupos de larvas de peces estuarinos varían continuamente en
espacial, de acuerdo con las estaciones reproductivas de la especie y debido
a
(Barletta-Bergan et al., 2002, Ramos et al.,
2006a; Montoya-Maya y Strydom, 2009; Strydom, 2015). Por consiguiente,
las condiciones ambientales pueden interferir con la
evaluación de calidad cuando se utilizan larvas de peces como indicadores
de
así como los patrones de variabilidad temporal asociados con fluctuaciones
naturales
de las estaciones reproductivas específicas de las especies, considerada
conductor de las asociaciones de peces larvales estuarinos (Rakocinski et
al., 1996).
La paradoja de la calidad estuarina (Elliott y Quintino, 2007), que
sugiere que el estrés antropogénico es difícil de detectar en los estuarios
porque
comunidades existentes también muestran la variabilidad natural,
se ha reflejado en muchos estudios de evaluaciones de la calidad ecológica
(p.
Courrat et al., 2009; Dauvin y Ruellet, 2009; Ramos et al., 2015;
Tweedley et al., 2015). Este es un problema particular en los estuarios desde
sus condiciones ambientales naturales varían espacialmente y temporalmente
(Sheaves, 2016). Así, la constancia de las características ambientales
es fundamental para las comparaciones espaciales de la categoría y
nivel de las presiones antropogénicas, eliminando las potencialmente
confusas
efectos de la variabilidad natural en las asociaciones de peces (Elliott y
Quintino, 2007). En este contexto, probamos la hipótesis de que los estuarios
con características medioambientales similares, pero distintos
presiones, llevan asociaciones de peces larvales claramente diferentes,
investigando
las asociaciones larvales de peces de cuatro estuarios tropicales similares
con diferentes tipos y niveles de presiones humanas.
2. Material y métodos
2.1. Enfoque metodológico
Para probar la hipótesis de que los estuarios con
características que sufren de presiones antropogénicas específicas
distintos ensambles larvales de peces, se seleccionaron cuatro estuarios
en la misma área biogeográfica de SE Brasil, con el fin de eliminar el
ensamblaje
diferencias debido a la localización geográfica de los estuarios. En
Además, todas las muestras se recogieron en la misma masa de agua (salinidad
15-25) para garantizar parámetros similares de agua natural que permitan la
interestuarina
Comparaciones. Teniendo en cuenta que las agrupaciones de peces larvales
estuarinos
son muy estacionales (Ramos et al., 2006b, Arévalo-Frías y
Mendoza-Carranza, 2015; Strydom, 2015), se analizó la lluvia / seco
temporadas por separado, asegurando que las posibles diferencias de
no relacionadas con influencias estacionales.

Por lo tanto, hemos restringido inter-estuarino

variabilidad en términos de condiciones ambientales naturales, asegurando
que
diferencias entre los estuarios se relacionaron con el tipo y nivel de
presiones antropogénicas. Las presiones antropogénicas que actúan
estuarios fueron evaluados en base a indicadores de contaminación, como
nutrientes
carga (nitrato, nitrito, amoníaco y fosfato) y contaminación de las aguas
residuales
determinada por la contaminación microbiana de coliformes fecales.
También se calculó un índice de presión para cada estuario basado en
actividades y presiones hidro-morfológicas.
2.2. Estuarine modelos
Cuatro estuarios de SE Brasil, Macaé, São João, Bracuí y Perequê-
Açu (Fig. 1). Macaé (22 ° 22 'S y 41 ° 46' W) y São
João (22 ° 35 'S y 41 ° 59' W) son sistemas más grandes directamente
conectados a
el océano con 130 km y 120 km de longitud, respectivamente; y total
área de 1765 km2 y 2160 km2, respectivamente. Bracuí (22 ° 57 'S y 44 °
23 'W) y Perequê-Açu (23 ° 13' S y 44 ° 42 'W) son sistemas más pequeños
con 32 km de longitud y una superficie total de 190 km2, y 22 km y área de
110 km2, respectivamente; conectado a una bahía costera (Bahía de Ilha
Grande).
Los cuatro estuarios son poco profundos (profundidad media de 2-4 m), tipo
de canal,
microtidal y parcialmente mixto, con régimen semidiurno.
Los niveles de presiones antropogénicas varían entre los estuarios. En
Macaé y Perequê-Açu, el proceso de urbanización no planificada
promovió la remoción de manglares en la región estuarina y la descarga
grandes cantidades de materia orgánica y otros compuestos de
residuos sin tratamiento directo en el agua (INEA, 2014, 2015).
Ambos estuarios se utilizan para la navegación, principalmente los barcos
de excursión en Perequê-
Açu, que se basa en el dragado constante para mejorar la navegabilidad.
El estuario de Macaé se utiliza como ruta de navegación para buques de apoyo
plataformas de perforación de gas. La entrada de agua dulce del río Macaé
es
controlado por la captación de agua para consumo humano, uso industrial
y enfriar el equipo generador de energía en plantas termoeléctricas
(INEA, 2014). Además, existen varias represas, actividades agrícolas
y extracción comercial de arena a lo largo del río Macaé. São João
Estuario todavía mantiene áreas de mangle en las secciones media e inferior,
aunque hay actividades agrícolas y pesqueras que pueden representar
una presión potencial. Además, el río principal está obstruido por una
, que controla el flujo del río que llega al estuario de São João.