Study on the consumption of microplastics by fishes in a floodplain lake of the Curiaú River (Macapá – Amapá, Brazil)

doi:10.21203/rs.3.rs-6497873/v1

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Study on the consumption of microplastics by fishes in a floodplain lake of the Curiaú River (Macapá – Amapá, Brazil)

https://doi.org/10.21203/rs.3.rs-6497873/v1

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published 09 Jul, 2025

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Plastics are used in almost every aspect of modern life from construction and electronics to clothing and food packaging. Freshwater ecosystems are the ultimate destination for many pollutants including plastic particles smaller than 5 mm, so-called microplastics. When available in the aquatic environment, these particles are actively or passively consumed by fishes. Studies on the natural diet of fishes can highlight and elucidate the impacts of this pollutant on aquatic ecosystems. The stomachs, intestines and gills from 122 fishes collected from Curiaú River Resort during the dry and rainy seasons, were analyzed by chemical digestion with KOH to verify the presence of microplastics. The fishes were categorized by taxa (14 species), feeding guild (herbivore, carnivore, piscivore, omnivore) and collection period (dry vs. rainy). We found a total of 732 microplastics, all classified as fibers, in 96% of the fishes examined. The predominant colors of the fibers were blue (59%) and black (33%). The highest consumption of microplastics occurred during the rainy season. Among the six most abundant species sampled, microplastics were most common in Geophagus sp., a fish that forages by sifting substrates dominated by sand. We also found differences between feeding guilds with carnivores scoring highest in the consumption of microplastics. We found no association between fish size and weight and the quantity of microplastics consumed. This study provides valuable baseline data on the ingestion of microplastics by fishes in the Curiaú Resort, and new insights into microplastic consumption by freshwater fishes. Our results are compared to similar studies of fishes in aquatic environments around the world.

Plastic pollution

Microplastic

Trophic guild

Microplastic ingestion

Freshwater fishes

Plastics are recognized as artificial substances composed of synthetic material or semi-synthetic natural polymers manufactured from petroleum-based chemicals. They are economical, lightweight, durable and corrosion resistant. (Boucher and Friot 2017). Plastics are used in almost every aspect of modern life from construction and electronics to clothing and food packaging. Plastic particles smaller than 5 mm are called microplastics (Hartmann et al. 2019) and generally result from the degradation of plastic waste. Microplastics can be categorized according to their source of origin: primary microplastics, for example, are used in personal care products, and secondary microplastics result from the degradation of larger pieces (Barnes et al. 2009).

Freshwater ecosystems are the ultimate destination for many pollutants resulting from the improper disposal of solid waste by the watershed’s human population. This solid waste is transported by rain and wind until it reaches water bodies. (Faure et al. 2015). Once in the water, plastics are transported by rising waters into floodplains or downstream by currents and can become trapped in riverbed sediments and structures (e.g. banks, bushes, trees and cliffs). (Azevedo Santos et al. 2021). In the Brazilian Amazon, plastic represents 15.7% of total solid waste (MMA 2015), and it is estimated that 182,085 metric tons of plastic are dumped annually into water bodies (Giarrizzo et al. 2019). Much of this waste is transported by the Amazon River to the Atlantic Ocean, making the Amazon the second most plastic-polluted river in the world, behind only the Yangtze River in China. (Giarrizzo et al. 2019).

Studies on the natural diet of fishes can highlight and elucidate the impacts on aquatic ecosystems resulting from dams, effluent discharge, and exotic species, among others. More recently, studies on fish diets have been used to detect the presence of microplastics in the environment and characterize the greater susceptibility of trophic groups to this new type of pollution. The objective of this study is to investigate 1) ingestion of microplastics by fishes in the Curiaú River resort area, located in a large floodplain lake in the municipality of Macapá (AP), and 2) potential associations between this ingestion and the biological characteristics of the fishes, such as species, feeding habits and body size.

This study took place at the Curiaú River Resort (Fig. 1) in areas close to places where bathers visit throughout the year, but especially during the summer months (July to December). The study area is in the Curiaú River Environmental Protection Area (APA do Curiaú) located in the urban expansion area of the Municipality of Macapá, state of Amapá, Brazilian Amazon. (Chellappa et al. 2005) in the vicinity of the coordinates 0°8'44.07"N, 51°2'31.58"W (Map 1). The Curiaú APA covers a small area (23,000 ha) in contrast to its high diversity of ecosystems such as savannas, dry-land forests, flooded forests and floodplains. The Curiaú River is a floodplain-system, with a dry (low-water) season occurring from July to December, and a rainy (high-water) season from January to June. (Chellappa et al. 2005).

Fishes were collected by residents of the Curiaú APA, in October 2022 and April 2023, corresponding to the dry and flood periods, respectively. Study specimens were donated to the Amapá Fauna Scientific Collection based at the Amapá Institute of Scientific and Technological Research (IEPA).

The gills and gastrointestinal tract (GIT) from the upper part of the esophagus to the anus were removed from each study specimen after taxonomic identification. The stomachs were then separated from the intestines and analyzed under a stereoscopic microscope to identify the feeding habits of each species. All samples (gills, stomachs, intestines) were transferred to individual glass beakers and taken to the Food Laboratory of the IEPA Food Science and Technology Center. There the samples were treated with a 10% potassium hydroxide (KOH) solution and the beakers were covered with aluminum foil to prevent possible contamination and evaporation. The samples were then heated at a temperature of 60°C for 24 hours (Suwartiningsih et al. 2020), which according to Dehaut et al. (2016) is the most effective protocol for the digestion of organic matter and preservation of microplastics (MPs). After digestion, the remaining solution was poured through 50 µm pore size filters in the IEPA Ichthyology Laboratory. The filters were placed onto sterile petri dishes for microscopic examination of microplastics. Finally, the plastics were visually assessed and categorized by color, size, and shape (i.e., fiber, fragment, line, film, or foam).

All laboratory procedures involved necessary precautions to avoid possible contamination of the samples. Laboratory personnel always wore nitrile gloves and cotton lab coats during analysis. Laboratory surfaces and digestion equipment were cleaned with pure water before and after each dissection. Before use, filters were inspected under the microscope for the existence of MPs. Control blanks were made for each day of analysis before beginning the sample digestion. For control blanks, a beaker was filled with 50 ml of the same KOH solution and covered with aluminum foil; these blanks were exposed to the same protocol applied to the samples.

Data were analyzed by species, feeding guild, and collection period. Microplastic values were subjected to the Shapiro-Wilk normality test and homoscedasticity of variances (Levene) (Sokal and Rohlf 1995) according to the most abundant species, guilds and collection period. Since microplastic values did not meet normality, the Kruskal-Wallis nonparametric test was used to compare the most abundant species and guilds, and the Kolmogorov-Smirnov test was used to verify differences between collection periods with a 95% confidence interval. Spearman correlations were performed between total length and weight and the amount of microplastics found for the most abundant species. All analyses were performed with the Statistica 7.0 program. StatSoft, Inc. (2004). STATISTICA (data analysis software system), version 7. www.statsoft.com.

Our analysis involved a total of 122 wild-caught fishes distributed among 14 species (Table 01); 92 and 30 specimens were collected during the dry and rainy seasons, respectively. Microplastics were not found in only five specimens representing the species Hoplias malabaricus (1), Metynnis lippincottianus (3) and Satanoperca jurupari (1), all from the dry-season collecting efforts. A total of 732 microplastics, all classified as fibers, were found in the 117 remaining specimens.

Table 1

Number of specimens (n) and mean ± standard deviation (M ± SD) of the amount of microplastics per species and their feeding habits (guild)
Species	n	M ± SD (cm)	Guild
Acestrorhynchus altus	2	8,00 ± 0	Piscivore
Aequidens sp.	2	3,50 ± 0,71	Onivore
Bryconops melanurus	15	8,80 ± 3,84	Carnivore
Crenicichla johana	2	2,50 ± 0,71	Carnivore
Crenicichla sp.	3	12,00 ± 14,73	Carnivore
Creniciclha cf. johanna	4	6,25 ± 1,26	Carnivore
Geophagus sp.	10	10,30 ± 5,31	Onivore
Heros severus	1	2,00 ± 0	Onivore
Hoplias malabaricus	2	5,50 ± 7,78	Piscivore
Mesonauta acora	9	7,33 ± 5,74	Onivore
Metynnis lippincottianus	39	3,69 ± 2,74	Herbivore
Satanoperca jurupari	24	4,04 ± 2,65	Onivore
Serrasalmus maculatus	1	5,00 ± 0	Onivore
Acestrorhynchus falcirostris	8	8,38 ± 4,14	Carnivore

Fibers of various colors were found in the three samples (gill, stomach, intestine): blue (59%), black (33%) red (6%), green (1%), and lilac (1%). The average size of the fibers was 0.27 cm, with green fibers being the largest (avg. 0.33 cm), followed by lilac (0.30 cm), blue (0.27 cm), red (0.24 cm) and black (0.23 cm) fibers.

Based on stomach contents, the species analyzed exhibited four different feeding habits (guilds): herbivore, carnivore, piscivore and omnivore. Microplastics were found in all guilds (Table 2) during both collection periods (dry and wet seasons).

Table 2

Number of specimens and mean ± standard deviation of the amount of microplastics by feeding habits (Guild)
Guild	n	M ± SD (MP)
Carnivore	30	8,67 ± 5,33
Herbivore	39	3,69 ± 2,74
Omnivore	49	6,04 ± 4,78
Piscivore	4	5,25 ± 3,20

Microplastics occurrence was highest (76%) in samples from the digestive tract (46% stomach and 30% intestines) with only 24% in samples from the gills.

Fish collected during the rainy season ingested a higher quantity of microplastics (p < 0.05) when compared to fish from the dry season (Fig. 1).

For the six most abundant species, the Kruskal-Wallis test identified two groups (a, b) with significant differences (p < 0.05) between them in relation to microplastic consumption in the study area (Fig. 2). Group a species (Bryconops melanurus, Geophagus sp. and Acestrorhynchus falcirostris) differed significantly from those of group b (Metynnis lippincottianus and Satonoperca jurupari). Mesonauta acora spanned both groups (a, b) and thus did not differ from the other species.

For the four different feeding guilds, the Kruskal-Wallis test identified three groups (a, b, c) with significant differences (p < 0.05) in microplastic consumption occurring between carnivores (b) and the herbivores and omnivores (c) (Fig. 3). Piscivores (a) did not significantly differ from any other guild.

The analysis of the relationship between microplastic consumption and fish weight (R²=0,0032, P < 0,05) and length (R²=0,0392, P < 0,05) was not significant for all specimens (n = 122), showing that the body size does not influence the amount of microplastics ingested.

All microplastics found in the three samples analyzed (gills, stomach, and intestine) were classified as fibers. Similar studies worldwide have found fibers to be the dominant type of MP contaminant in fishes (Jabeen et al. 2016; Gomez et al. 2020; Pappoe et al. 2022; Kiliç et al. 2022, Bellas et al. 2016, Chan et al. 2019, Justino et al. 2021), suggesting that fibers are the most abundant microplastics ingested in aquatic environments.

The high frequency of microplastics in the current study (found in 95.9% of 122 individual fishes) was similar to those reported by Aunurohim et al. (2023) (99% of 74 individuals) and Suwartiningsih et al. (2020) (97.5% of 80 individuals), but higher than those of other fish diet studies such as Khan and Setu (2022) (76% of 45 individuals) and as Vendel et al. (2017) (9% of 2233 individuals) and some others studies as shown in Table 3. This discrepancy can be explained in part by our use of the protocol developed by Dehaut et al. (2016) which highlights small plastic particles that cannot be observed in situ (i.e., within the digestive tract and its contents). Microplastics also adhere to the mucus of gill filaments, a fact that makes it extremely difficult to visualize them, even under a microscope. The chemical digestion of organic matter leaves only the inorganic matter visible, such as ingested sediments and synthetic materials, highlighting the occurrence of microplastics.

Microplastics found in both marine and freshwater environments can be transparent or display a wide variety of colors, including black, blue, gray, green, red, white, purple, or yellow (Wang et al. 2021). Experimentally, it has been observed that some fish species prefer ingesting certain colors of plastics, and the prevalence of different colors of microplastics can vary widely across sampling sites (Horie et al. 2024). The predominance of blue and black fibers in the analyzed fishes follows a general pattern of occurrence found in other similar studies. For example, the most common colors were black, white and blue among microplastics reported by Khan and Setu (2022) in freshwater fishes in Bangladesh, black in demersal fish from the Spanish Coast and Mediterranean Sea (Bellas et al. 2016) and pelagic and demersal fish in Indonesia (Aunurohim et al. 2023), and black, blue and red in serrasalmids from Xingu River, Brazil (Andrade et al. 2019).

The occurrence of microplastics in the three different samples from each individual (gill, stomach and intestine), indicates that these particles contaminate the fish through two different pathways: one passive, with the accumulation of plastic in the mucus of the gills during swimming or breathing (Parker et al. 2021), and the other active, due to feeding activity, wherein ingestion may or may not be voluntary. Ingested plastics presumably return to the environment with the fishes’ feces, otherwise larger fishes would have a greater accumulation of fibers in their stomach and intestine. We found no correlation between fish body size and the ingestion of microplastics, as did Chan et al. (2029) in Hong Kong Coast and Vendel et al. (2017) in Paraiba and Mamanguape River estuary (Brazil). Suwartiningsih et al. (2020) found only a weak correlation in Yogyakarta, Indonesia, whereas Khan and Setu (2022) found body size to be positively correlated with microplastic abundance in fish specimens from the Jamuna River, Bangladesh. Our analyses were unable to determine the proportion of ingested microplastics potentially absorbed into the bloodstream and incorporated into the fishes’ bodies.

Table 3

Comparison of microplastic occurrence in fishes. GIT = gastrointestinal tract, C = carnivore, H = Herbivore, O = Omnivore, P = Piscivorous, Z = Zoobenthivorous, I = Insectivore, Il = Iliophage, D = Detritivore, A = Algae eaters, Zo = Zooplanktivores, B = Benthivores, G = Generalists, Pl = Planktivorous
Location of Study	Habitat	No. of fish species	Trophic Guilds	Samples	Method	Total No. of Individuals Examined (no. with microplastics)	% with Microplastics	Study
Indonesia	Marine	11	-	GIT	KOH	75 (74)	99%	Aunurohim et al. 2023
Indonesia	Marine	4	-	GIT	KOH	80 (78)	97.5%	Suwartiningsih et al. 2020
Brazil	Freshwater	14	C, H, O, P	Gill, GIT	KOH	122 (117)	95.9%	this study
Bangladesh	Freshwater	7	H, C, O	GIT	H₂O₂	45 (34)	76%	Khan and Setu 2022
Brazil	Brackish	3	D, Z, P	GIT	NaOH	82 (69)	73%	Justino et al. 2021
Ghana	Marine	4	-	GIT	KOH + H₂O₂	115 (79)	68.7%	Pappoe et al. 2022
Brazil	Marine	7	O, Z	Stomach	Stereomicroscope	214 (118)	55%	Dantas et al 2020
China	Marine, brackish	4	-	Stomach	HNO₃	147 (80)	54%	Chan et al. 2019
Brazil	Freshwater	16	C, H, O	Stomach	Stereomicroscope	172 (46)	26.7%	Andrade et al. 2019
Spain	Marine	7	-	Stomach	NaOH	212 (37)	17.5%	Bellas et al. 2016
Philippines	Marine	-	-	GIT	NaOCl	180 (21)	11.67%	Gomez et al. 2020
Brazil	Brackish	69	A, Zo, Z, B, G	GIT	Stereomicroscope	2233 (196)	9%	Vendel et al. 2017
Panama, Colombia, Ecuador, Peru, and Chile	Marine	7	Pl	GIT	Stereomicroscope	292 (6)	2.10%	Ory et al. 2018
Brazil	Freshwater	14	O, I, P, Il	Stomach	Stereomicroscope	220 (4)	1.8%	Oliveira et al. 2020

Dantas et al. (2020) reported that microplastic ingestion does not depend on eating habits of marine fishes in Ceará State (Brazil); however, they analyzed only two trophic guilds (see Table 3). Khan and Setu (2022) also found no significant differences between feeding habits and microplastics ingestion. Nevertheless, other studies have shown that microplastic ingestion varies according to feeding strategy (Table 3). Ismail et al. (2018) reported that herbivorous fishes of Biawak Island showed a higher density microplastics, while Andrade et al. (2019) found small differences in MP intake patterns between fish guilds observed in the Xingu River. In the current study, we observed a significant difference between feeding habits and microplastic consumption, suggesting that piscivorous fishes ingest not only the MPs of their prey, but MPs taken incidentally during prey capture (Justino et al. 2021) and respiration. Parker et al. (2021) indicated that the ingestion of microplastics by fish may be related to their feeding activity, which would be directly influenced by environmental conditions that determine the prey’s abundance and type (Jobling 1981). Habitats that concentrate or receive a lot of microplastics also increase the chance of microplastics being ingested (Wright et al. 2013; Güven et al. 2017).

In the environment, MP deposition and accumulation patterns may vary by substrate (i.e., sediments vs. plants) and in the water column. These patterns influence the amount of MP ingested by fishes according to their diet and feeding habits (on the bottom vs. in the water column, as for piscivores). Accordingly to our results, piscivores and omnivores occupy an intermediate position (Fig. 9) between carnivores and herbivores. This also explains the higher consumption of microplastics among fishes collected during the high-water season, a period when food availability is greater and consequently fish increase their feeding activity. In addition, the dry (low-water) season can affect the availability of MP in the water. Reduced flows and shallower depths cause microplastics to accumulate in the banks and substrate, thereby decreasing their availability in the water (Han et al. 2020). During the rainy season, rainwater becomes an important carrier of MP, as it removes MP from the atmosphere causing the phenomenon known as plastic rain (Brahney et al. 2021). Thus, the increase in rainwater and flooding during the rainy season increases the amount of microplastics suspended in the water column. All this emphasizes that hydrological changes affect not only the quantity, but also the quality of the food (Oliveira et al. 2020).

This study showed that the Curiaú Resort is affected by microplastic pollution, and that fish regularly consume these particles. It can be concluded that plastic pollution in the waters of the Curiaú River is already impacting the aquatic fauna and that this problem should carefully monitored. Not only can these particles cause diseases and problems for fishes, but these fishes are part of the diet of much of the population of Amapá State and this contamination can affect their health as well.

This study provides valuable baseline data on the ingestion of microplastics by fishes in the Curiaú Resort, a protected area frequented by tourists and fishermen and with great social and ecological value.

Conflict of interest

The authors declare that they have no potential conflict of interest to disclose.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Author Contribution

CSG contributed to the study conception and design. Material preparation, data collection and analysis were performed by LMM, TCS, AAP and KRR and supervised by CSG. Thes statistical analysis and interpretation were carried by LMAS and LMM. The digestion of the samples was supervised by CSG and ACFS. The first draft of the manuscript was written by CSG and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Acknowledgement

We would like to thank Dr. Mark Sabaj for his insights and help reviewing this manuscript and to Andrio L. M. de Souza for his help in creating the map.

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Map 1 is available in the Supplementary Files section.

No competing interests reported.

Map1.jpg
Map 1- Location of the study area.

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26 Apr, 2025
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21 Apr, 2025

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Study on the consumption of microplastics by fishes in a floodplain lake of the Curiaú River (Macapá – Amapá, Brazil)

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Journal Publication

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Abstract

Figures

Introduction

Methodology

Results

Discussion

Conclusions/final considerations

Declarations

Conflict of interest

Funding

Author Contribution

Acknowledgement

References

Map

Additional Declarations

Supplementary Files

Status:

Journal Publication

Version 1