Next Article in Journal
Surface Flux Patterns of Nutrient Concentrations and Total Suspended Solids in Western Carpathian Stream within Agricultural, Forest, and Grassland Landscapes
Previous Article in Journal
Comparison of Hexavalent Chromium Adsorption Behavior on Conventional and Biodegradable Microplastics
Previous Article in Special Issue
Unraveling the Sea Slug Fauna from an Extremely Variable Environment, The ‘Passetto’ Rocky Tide Pools (North Adriatic Sea)
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Water
Volume 16
Issue 14
10.3390/w16142051
water-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Influence of Temperature on the Toxic Effects of Carbamazepine on the Copepod Tigriopus fulvus: A Transgenerational Full Life Cycle Study

by
Isabella Parlapiano
,
Ermelinda Prato
*,
Giuseppe Denti
and
Francesca Biandolino
National Research Council, Water Research Institute (CNR-IRSA), 74123 Taranto, Italy
*
Author to whom correspondence should be addressed.
Water 2024, 16(14), 2051; https://doi.org/10.3390/w16142051 (registering DOI)
Submission received: 15 May 2024 / Revised: 15 July 2024 / Accepted: 17 July 2024 / Published: 19 July 2024
(This article belongs to the Special Issue Implementation of Biodiversity and Ecosystem Services in Marine Ecosystem Management, 3rd Edition)
Browse Figures
Versions Notes

Abstract

:
Coastal areas are increasingly exposed to global warming and emerging contaminants from anthropogenic activities; however, the interactive effects of these stress factors in shaping the offspring’s vulnerability to them are poorly understood. The present study aimed to assess the influence of temperature on the toxicity of the pharmaceutical carbamazepine (CBZ) in the parental (F0) and in the first (F1) generation of Tigriopus fulvus, through a full life cycle study, measuring several biological parameters. At control temperature (20 °C), exposure to CBZ significantly inhibited larval development, especially in the F1 generation. In contrast, under warmer conditions (27 °C), even after exposure to CBZ, the development was stimulated, proving that temperature was the main factor influencing it. As regards the other investigated life traits (body length, sex ratio, and fecundity), both temperature and generation modulated toxic effects of CBZ, which is evidenced by the onset of higher alterations in F1 co-exposed copepods. Our findings suggest that temperature and contaminants could increase the long-term vulnerability to stressors of T. fulvus, potentially affecting the population structure over multiple generations of exposure.

1. Introduction

Pharmaceuticals over the last two decades have been receiving increasing attention from the scientific community due to their ubiquitous presence in all environmental compartments [1]. The continuous detection of such compounds in water bodies and the evidence of detrimental biological effects related to exposure resulted in their designation as contaminants of emerging concern (CECs); however, routine monitoring programs, at the moment, do not include them, but may be eligible for future legislation depending on their ecotoxicity [2].
Pharmaceuticals include very complex molecules with different physicochemical properties. Even the most degradable ones are considered persistent or pseudo-persistent compounds because they are constantly discharged into the aquatic environment [3], leading to a scenario of pseudo-chronic exposure to non-target organisms. Considering that pharmaceuticals are capable of being active in small doses, they may exert adverse long-term effects on non-target aquatic organisms systematically far from target organisms, through molecular and metabolic pathways evolutionarily conserved, even at very low concentrations [4].
In marine ecosystems, despite the high dilution factor, pharmaceutical compounds can be found at levels between a few ng/L up to hundreds of µg/L [5,6], posing a growing risk for coastal zones [7].
In addition to anthropogenic pollution, coastal ecosystems are subjected to physicochemical stressors related to climate change more pronounced compared to the offshore ones [8]. It is estimated that by the end of the XXI century, the surface temperature in certain coastal zones of the Mediterranean Sea will increase by 4 °C, compared to the 0.6–2 °C predicted in the global ocean [9,10], and there will be at least one long-lasting marine heat wave event every year, up to three months longer, about 4 times more intense and 42 times more severe than current events [10].
The vulnerability of shallow and intertidal coastal habitats to these extreme events may lead to an increase in thermal stress in these environments.
There is a growing amount of research revealing that factors related to global warming can modulate the bioavailability and the effects of several pollutants in coastal environments [11]; anyway, the possible interactions between these co-stressors are still unclear [12,13]. Since ectothermic aquatic organisms are susceptible to changes in temperature because of their biology [14,15], global ocean warming is one of the most common stressors that may affect biological processes in marine species [16,17]; in addition, high temperatures cause a wide variety of contaminants to become more toxic [18]. A rise in temperature, for instance, can alter the transport and the chemical behavior of a substance, and at the same time increase the metabolic rate of organisms and the transport of toxic compounds through food items and cellular membranes, likely resulting in a higher intake of them and consequently a higher toxicity to organisms [19,20]. Therefore, the temperature may affect the traditional ecotoxicological risk assessment of chemicals generally based on the responses obtained with standard toxicity tests under constant and favorable experimental conditions.
To evaluate the influence of increased temperatures on the possible alteration of toxicity of pharmaceuticals, we selected one of the most consumed in the world: the anti-epileptic agent carbamazepine (CBZ) [21]. Because CBZ is lipophilic, resistant to photodegradation and biodegradation, it is also minimally removed in modern multi-stage sewage treatment processes, has a long half-life [22,23], is poorly degraded, and remains in the environment for a long time, leading to its widespread occurrence in the water column up to tens of μg/L and in the tissues of aquatic invertebrates up to 280 ng/g (d.w.) [24,25,26]. For these reasons, CBZ has been proposed as a marker for anthropogenic contamination [27]. Furthermore, the potential interactions of co-occurring stressors on marine organisms were recently confirmed for pharmaceuticals including CBZ [28,29,30,31]. Many studies have demonstrated that CBZ at environmental concentrations affects several physiological, behavioral, and reproductive processes in a variety of non-target aquatic organisms including fish and invertebrates, with significant deleterious effects [32,33,34,35,36,37]; thus, the European legislation on the classification and labeling of chemicals (92/32/EEC) classified CBZ as “R52/53 Harmful to aquatic organisms and may cause long-term adverse effects in the aquatic environment”. As regards marine crustaceans, a recent study has demonstrated that CBZ significantly affected the expression of biomarkers of exposure of the crab Carcinus maenas, [38] and inhibited molting and growth of the shrimp Eriocheir sinensis [39]; anyway, very limited information is available on the chronic effects of CBZ on full life cycle traits of marine crustaceans.
In a recent study [40], the chronic toxicity of CBZ was assessed in a broad range of environmental concentrations under standard test conditions on the marine meiobenthic copepod Tigriopus fulvus (Fisher, 1860). Due to its biological features (small size, short life cycle, high fecundity, distinct post-embryonic stages, sexual dimorphism), T. fulvus represents an ideal marine test organism for water quality assessment [40,41,42,43,44,45,46,47]. This copepod is widely distributed along the Mediterranean coasts in shallow supratidal rocky pools; therefore, it is a species subjected to large environmental fluctuations that favor its high adaptability and tolerance [48], so it can be used in ecotoxicological studies under a wide range of environmental conditions. As a generalist feeder, it is an important energy link in marine food webs, and it plays an important role in transferring aquatic pollutants across the food chains [49]. Several studies demonstrated that species belonging to the genus Tigriopus are also suitable for use in chronic tests through a multi-generational approach [50,51,52].
It is becoming increasingly evident that environmental conditions, such as warming and/or contaminants experienced by parents, may influence the performance of multiple generations of offspring in a positive or negative way [53,54,55,56]; however, studies on the interactions of co-stressors on successive generations are still scarce. Therefore, the objective of this study was to measure the effects of CBZ on the survival, development, growth, sex ratio, and reproduction across two generations of T. fulvus under control temperature test conditions (i.e., 20 °C) and warmer conditions (i.e., 27 °C), at a temperature 4 °C above the surface maximum average value registered in summer over a thirty-year period in the northwestern Mediterranean Sea in T. fulvus habitat [10]. Extreme heat wave events with a peak of 27 °C have been observed along the coast in recent years and are known to cause mass mortality events in marine invertebrates [57].
The life traits investigated are key parameters for copepod population recruitment and structure. Our initial tested hypotheses were as follows. (1) The highest temperature increases the direct effects of CBZ on T. fulvus with respect to the optimal temperature; (2) the parental exposure to CBZ and temperature shape offspring vulnerability to these stressors, resulting in either enhanced toxicity or resistance in subsequent generations.

2. Materials and Methods

2.1. Test Organisms and Chemicals

T. fulvus used in all experiments came from laboratory cultures established for several months in the CNR Water Research Institute of Taranto, Italy, according to the conditions indicated by the UNICHIM 2396:2014 method [58].
A stock solution of carbamazepine (CBZ) (100 mg/L) (Sigma-Aldrich, Saint Louis, MO, USA, CAS 298–46-4, purity > 97%) was prepared in HPLC-grade methanol for analysis (Sigma-Aldrich, Milano, Italy, CAS 32213-M purity ≥ 99.8%) and stored in amber vials at 4 °C until use. Stock solution was diluted in 0.22 μm-filtered artificial seawater (ASW) Instant Ocean® (IO) just before testing, adding the microalgae Tetraselmis suecica (105 cells/mL) as food source, to obtain working solutions, ensuring that methanol was first completely evaporated. Working solutions were renewed (>80% of the volume) every 48 h. Since CBZ is a very stable compound in aquatic environments, with a half-life of over 100 days [59], any slight changes in the concentration of the solutions during tests were considered negligible.

2.2. Test Procedures

According to previous studies [40,42,46], to start the parental-generation test (F0) a total of 72–78 newly hatched nauplii (<24 h) per treatment was randomly selected and distributed in six replicates into 12 multiwell plates (Nest Biotech Co., Ltd, Wuxi, China). Each replicate/well contained 12–13 nauplii in a volume of 4 mL of test solution. Plates were incubated under the same conditions as the stock culture (16 h light/8 h dark photoperiod, salinity of 38 PSU ± 1, dissolved oxygen saturation > 80%, pH of 8 ± 0.3) at two distinctive temperatures (20 °C control and 27 °C).
Copepods were exposed for 28 days to 10 µg/L of CBZ; the tested concentration was chosen based on the results of the previous life cycle experiment mentioned above, in which it was observed that most of the investigated endpoints were significant at 10 µg/L of CBZ [40].
Plates were observed every 24 h under a stereomicroscope with a scattering light Zeiss Stemi SV 11 (Jena, Germany) to determine the developmental stage and calculate the time required for nauplii to become copepodites (naupliar phase duration) and the time required for nauplii to reach the adult stage (i.e., females with ovigerous sac). After all copepods matured, survival (%) and sex ratio (female/male) were assessed. The survival rate was calculated as mean percentage of living nauplii reaching the copepodite and the adult stages at maturity. Moreover, at the end of the experimental period, the body length of male and female specimens was determined under an inverted microscope Zeiss Axiovert S100 (Jena, Germany) equipped with a camera Leica MC170HD with a 10× magnification using LAS X Leica Microsystem CMS GmbH software (Wetslar, Germany). To measure the fecundity (offspring production), six females with egg sac per concentration were individually transferred to a new 12-well culture plate in a volume of 2 mL of fresh solution per well. Plates were inspected daily to count the number of nauplii per female, the number of broods per female, and the unhatched broods per female, which is defined as the broods unable to produce viable offspring (i.e., aborted egg sacs), and to calculate the time required for the offspring release (hatching time).
Working solutions were changed every 24 h and nauplii removed under the stereomicroscope. The first-generation test (F1) started with nauplii produced by F0 females with the second or third brood; experimental conditions were identical to the F0 generation test. Each generation test was carried out three times.

2.3. Statistical Data Processing

Data were presented as means ± standard deviation (SD). The interactions between the three factors (CBZ × Temperature × Generation) on the dependent variables were not tested because the assumptions for performing the three-way ANOVA were violated.
For this reason, separate tests were applied on the F0 and the F1 generations and to compare the two generations.
Data were tested for normality and variance homogeneity in each generation with Shapiro–Wilk and Levene’s tests, respectively. If both assumptions were met, data were examined by analysis of variance (one-way ANOVA), and the Tukey’s post hoc test was used to find significant differences (p < 0.05) among treatments. If normality and/or homogeneity of variance failed, data were log-transformed. When data did not conform to the ANOVA assumptions, even after transformation, the non-parametric Kruskal–Wallis test followed by the Dunn’s post hoc test was run. Specifically, one-way ANOVA was used on untransformed data of survival, length of males and females, hatching time, F0 nauplii/Fov, and on log-transformed data of F1 nauplii/Fov; development, hatching time, brood/Fov, number of nauplii/brood, and unhatched brood were analyzed with the Kruskal–Wallis test.
Since the F1 offspring derived directly from the F0 females (and so generations were dependent), the non-parametric Friedman test for repeated measures followed by the Wilcoxon post hoc test was applied to evaluate the transgenerational effects on variable responses caused by exposure to CBZ at the two temperatures. The two different generations, being dependent, were considered as being a single group from which the repeated measures were taken.
All statistical analyses were performed using Past3 software (version 1.0).

3. Results

3.1. Survival

In both generation F0 and F1, no significant differences were observed in the survival in all treatments (p > 0.05); nauplii became copepodites and adults with survivorship > 95%.

3.2. Effects in the F0 Generation

At 20 °C, naupliar phase duration and age of ovigerous females were slightly prolonged in CBZ treatments compared to the control, although differences were significant only for naupliar phase duration. In particular, the time required for nauplii to become copepodites was 5.46 ± 0.50 days in the control and 5.66 ± 0.56 days in the CBZ group, while the time for the appearance of ovigerous females was 14.11 ± 0.68 and 14.50 ± 0.71 days, respectively. On the contrary, at 27 °C, copepod development was significantly accelerated; in fact, the production of copepodites and adult females occurred around the 4th and the 11th days, respectively, both in the control group and the CBZ group (Figure 1A).
Sex ratio (i.e., number of females/number of males) was influenced only by temperature; indeed, this parameter increased at the higher temperature in both treatments compared to the control temperature, although significantly only in copepods exposed to CBZ.
No differences in the body length of males and females exposed to CBZ were observed in the F0 generation with respect to the control at both temperatures (Figure 1C).
Hatching time was influenced only by CBZ, resulting in it being significantly prolonged both at 20 °C and at 27 °C. Hatching occurred after about 2.5 and 2.8 days in the controls and in the CBZ groups, respectively, at both tested temperatures (Figure 1D).
CBZ exposure at 27 °C negatively affected the fecundity; indeed, the mean total number of nauplii per female at the end of the experiments in the control was 119.3 ± 38.3, while in the CBZ group, it was significantly reduced by 27% to 86.4 ± 37.9 (Figure 1E).
The number of broods was 5.4 ± 0.6 and 6.1 ± 0.8 in the control groups and 4.8 ± 0.6 and 5.6 ± 0.7 in the CBZ groups, at 20 °C and 27 °C, respectively, highlighting a reduction in exposed groups, although significant only at 20 °C. The statistical analysis also evidenced a significant increase in the control and the CBZ group at 27 °C, compared to both treatments at the control temperature (Figure 1F).
A reduction of the mean number of nauplii per brood occurred only at 27 °C, where there were fewer nauplii per brood in both treatments (19.5 ± 5.6 in the control and 15.8 ± 6.6 in the CBZ group) with respect to the control temperature (21.8 ± 3.4 in the control and 22.6 ± 4.2 in the CBZ group), although the differences were significant only in the CBZ group (Figure 1G).
Temperature and CBZ exerted a significant effect on the number of aborted sacs. The mean number of unhatched broods per female, in fact, was significantly higher only in copepods exposed to CBZ at 27 °C with respect to the other treatments (Figure 1H).

3.3. Effects in the F1 Generation

As regards naupliar phase duration, in the F1 generation, it was observed that the same trend occurred as in the F0 generation, although the magnitude of the CBZ effect was more pronounced in the offspring; indeed, nauplii in the CBZ group at 20 °C became copepodites after 6.3 ± 0.9 days.
Differently from the F0 generation, a significantly slower maturation of females was also detected in the CBZ group at 20 °C in comparison to the control group; in fact, the mean age of ovigerous females was 15.4 ± 1.1 days (Figure 2A).
Temperature strongly influenced the sex ratio in the F1 generation as well as in the F0 one. The sex ratio significantly increased at the higher temperature in both treatments compared to the control temperature (Figure 2B).
Differently from the F0 generation, at 27 °C, the length of females was significantly reduced in the CBZ group, measuring 698.0 ± 25 μm, compared to the control group, measuring 814.7 ± 91.6 μm, and compared to 20 °C. Indeed, at the control temperature, females measured 870.7 ± 113 and 891.8 ± 43.7 μm in the control in the CBZ treatment, respectively (Figure 2C). Finally, as regards the number of unhatched broods, the same pattern was observed as in the F0 generation.
Unlike the F0 generation, both temperature and CBZ influenced hatching time. In fact, at 27 °C, in the F1 copepods exposed to CBZ, a significant increase was observed in the duration of hatching time (3.0 ± 0.4 days) compared to the copepods exposed to CBZ at 20 °C (Figure 2D).
Like in the F0 generation, CBZ exposure negatively influenced the mean number of nauplii per female, although the observed reduction (38%) at 27 °C was stronger compared to the control group (Figure 2E).
As regards the number of broods, unlike the F0 generation, responses at 27 °C were more pronounced, resulting in a reduction of 25% in the CBZ treatment with respect to the control, and an increase of about 30% in the control with respect to the control at 20 °C (Figure 2F).

3.4. Transgenerational Effects

The non-parametric test for repeated measures evidenced that at the control temperature, nauplii of the F1 generation exposed to CBZ became copepodites with a significant delay of about 0.6 days with respect to the copepods exposed to CBZ of the F0 generation. At 27 °C, both in the control group and in the CBZ group, naupliar phase duration was significantly prolonged with respect to the F0 generation (Table 1).
In the F1 generation, at 20 °C, the time for the appearance of ovigerous females exposed to CBZ was significantly prolonged by about 0.9 days compared to the F0 generation (Table 1), reflecting the delay observed in the duration of the naupliar phase. A slight delay of 0.5 days was also observed in females exposed to CBZ at 27 °C with respect to the F0 generation, although this was not significant (Table 1). Furthermore, at 27 °C, the body length of F1 females exposed to CBZ was significantly reduced by 22% compared to the F0 females exposed to CBZ.

4. Discussion

Most previous studies investigating the combined effects of contaminants and temperature as co-stressors have centered on short-term experiments not considering the relevance of chronic exposure, especially over multiple generations [60]. CBZ is known to exert deleterious effects on aquatic non-target species, affecting development and reproduction [39,40,61,62,63], and enhanced toxicity has been observed when combined with marine heatwaves in Mytilus galloprovincialis [28].
Concerning crustaceans, existing knowledge on the combined effects of CBZ and temperature is limited to the freshwater shrimp Atyaephyra desmarestii, in which CBZ, under increasing temperature conditions, at a concentration of 13.8 µg/L, may produce respiratory deficiencies causing alterations in its behavior [64].
In the present study, based on previous results obtained on the parental generation of the marine copepod T. fulvus at the control temperature (i.e., 20 °C) [40], we examined the toxic effects of CBZ on development and reproduction over two generations at two different temperatures. Generally, such results demonstrated that, except for the survival and the size of males, CBZ exposure and temperature—alone or in combination—negatively affected all the sub-lethal investigated life cycle traits of T. fulvus, with stronger effects in the F1 generation.
The transition from the naupliar stage to the copepodite stage and from naupliar stage to adult stage (i.e., females with ovigerous sac) was confirmed to be a sensitive endpoint [40,42,46]. At the control temperature, in F0 copepods exposed to CBZ, it was observed that a slight slowdown of development occurred compared to the control. Since CBZ is a potential endocrine disruptor, it could have affected the metamorphosis process that is necessary for growth and development. In this regard, Chen et al. [39,62] showed that exposure to 3 and 10 μg/L of CBZ can inhibit the molting process of Daphnia similis and Eriocheir sinensis, respectively, by interfering with the activity of chitinolytic enzymes and the signaling of molting hormone, causing a slowdown of development. In the present study, the delay observed in copepods exposed to CBZ in the F1 generation was significantly more pronounced with respect to the F0; nauplii became copepodites and adults with a delay of about 0.6 and 0.9 days, respectively. Considering that CBZ is a relative lipophilic (Kow = 2.2) and bioaccumulative compound [65,66,67], the increased retardation in maturation observed in the F1 generation with respect to the F0 generation could have been caused by a longer time of exposure to the pharmaceutical. Several studies on aquatic invertebrates revealed that pharmaceuticals bioaccumulated in the exposed organisms may be transferred through the mother to eggs and deposited in the offspring, leading to a transgenerational transfer of toxicity [68]. As regards the results observed under seawater warming conditions (i.e., 27 °C) for these life traits, temperature strongly affected the development of T. fulvus compared to the CBZ, appearing to be the primary factor that influenced it. Warming also significantly shortened the development time of T. fulvus after CBZ exposure, especially in the parental generation. This could be explained considering that marine copepods tend to develop faster to the reproductive age with the increase in temperature [69,70,71,72]. In accordance with our results, Li et al. [73] showed that the development time of the congener T. japonicus decreases with high temperature. In the F1 females exposed to CBZ, the acceleration of maturation was less noticeable, probably due to an enhanced absorption of CBZ in the offspring.
As regards sex ratio, according to previous studies on T. fulvus [40,42,46], a trend toward more females per male, which is quite common for harpacticoid copepods [74,75,76], was observed in all treatments. In general, the existing literature shows a relatively high variability of sex ratio in crustaceans under warmer conditions. For example, in the calanoid Paracartia grani reared under warmer conditions for several generations, sex ratio was close to 1, showing to be not influenced by temperature [77]; on the contrary, in the harpacticoid T. californicus, a sex ratio shift (more males per female) was observed with the increasing temperatures, even after several generations [78]. Our results showed that temperature alone affected sex ratio, which increased from a minimum of 1.1 at 20 °C to a maximum of 3.5 at 27 °C. Similarly, Koch et al. [79] showed an increasing ratio of females per male from 0.7 to 4.0 with increasing temperatures in the harpacticoid Nitokra spinites. The sex determination in response to environmental factors could be an adaptative mechanism which needs to be further investigated, since a highly biased sex ratio may influence the size and the reproductive output of a population [80].
Another life trait influenced by temperature was the body length of females. Female size at maturity could be an important parameter influencing fecundity of copepods, as observed by Horne et al. [81]. The results obtained evidenced that CBZ enhanced the negative influence of warming on growth. This was particularly evident in the F1 co-exposed females, which showed a size reduction by 14% compared to the control females at 27 °C and by 22% compared to the CBZ-exposed females at 20 °C. Our results confirm that under warmer conditions, copepods develop faster, achieving smaller sizes at maturity [82,83,84], with consequences for their role on trophic web and community structure. For example, the increase in mean temperature recorded in an estuary on the Atlantic coast of the USA caused a decrease in body size of two species of the genus Acartia, the disappearance of two species of large copepods, and an increase in the proportion in the community of the small copepod Oithona sp., leading to an alteration in community composition [85]. In addition, smaller organisms are more sensitive to chemicals, which in turn can cause a decrease in body size through a variety of mechanisms [82]. Alterations in development and growth may be accompanied by impairment of other physiological traits, such as fertility.
In the present study, the fecundity-related traits of T. fulvus exposed to CBZ (i.e., number of broods/females, number of nauplii/broods and number of nauplii/females) were significantly reduced compared to the control in both generations.
The toxicity of CBZ was modulated by temperature, as evidenced by the onset of greater alterations caused by combined stressors rather than single stressors, but also by generation. Overall, after 28 days’ exposure, the most severe effects on reproduction were observed in F1 co-exposed females, in which the mean number of nauplii and broods decreased by 38% and 25%, respectively, compared to the control females, evidencing that the combined effect of both stressors may lead to greater toxicity on subsequent generations.
At environmental levels, CBZ has been proven to cause oxidative status alterations, lipid peroxidation, immune system impairment, and genotoxic damage in marine invertebrates [28,36,67]. This may diminish the investment of energy for reproduction, because of the increased energy costs for defence, detoxification, and repair mechanisms, as is well documented in cladoceran freshwater crustaceans [86,87].
Previous studies showed that the reproductive output of Daphnia similis [62], D. magna [61], D. pulex [88], and Ceriodaphnia dubia [89] significantly decreased after chronic exposure to CBZ at low concentrations ranging from 0.03 μg/L to 200 μg/L. Chen et al. [62] demonstrated that CBZ may act as an endocrine disrupter in D. similis, interfering with the endocrine system, which regulates various physiological functions in crustaceans, such as development, growth, sex ratio, and reproduction. In accordance with our results, in a multigenerational study, Nkoom et al. [63] found that exposure to 2 and 10 μg/L of CBZ significantly inhibited growth, molting frequency, and reproduction, and induced significant behavioral changes in D. magna, in the parental as well as the first generation, with relatively greater effects in the first one. Furthermore, the simultaneous reduction in the expression of molting, as well as reproductive and antioxidant-related gene expression, suggests that this pharmaceutical may cause oxidative stress and act as an endocrine disruptor. In another recent study, He et al. [90] reported that 5 μg/L of CBZ altered the expression of genes related to reproduction and toxic metabolism in two generations of D. magna, but that the transcriptional changes at the gene level were not fully translated into physiological performance in the F0 generation, while they were translated into a significant reduction in offspring in the F1 generation. According to these results, our findings also suggest that exposure to CBZ could act as an endocrine disruptor in T. fulvus and could also determine genetic alterations, as evidenced by the occurrence of greater effects in the first generation.
Hatching time was significantly extended in copepods exposed to CBZ with respect to the control, both at 20 °C and 27 °C in both generations, with a maximum increase of 0.5 days in F1 co-exposed females, but no differences occurred between the two generations. The prolonged hatching time indirectly affected fecundity, resulting in fewer broods. Finally, copepods aborted their egg sacs in response to both stressors. Specifically, F0 co-exposed females aborted the highest number of sacs, while in the F1 co-exposed females, there was a lower disturbance of temperature, suggesting a greater incidence of the pharmaceutical.
Overall, results reported here showed a greater incidence of the pharmaceutical on reproductive outcome, which could be the consequence of its greater bioaccumulation through the maternal transfer in the offspring [68].
At the same time, our results provide clear evidence of the capability of temperature to increase T. fulvus sensitivity to CBZ. Indeed, although, under warmer conditions, copepods began to reproduce earlier, the major inhibition of fecundity occurred in co-exposed females. This could be explained considering that a higher temperature increases the transport and uptake of contaminants across cell membranes, in response to an increased metabolic rate, which can result in the enhanced sensitivity of exposed organisms [91].
Recent studies on aquatic invertebrates highlighted the role of parental exposure to warming and contaminants in shaping the susceptibility of offspring to stressors. In the mosquito Culex pipiens, the interaction between warming and the insecticide chlorpyrifos was synergistic in both generations, despite the increased tolerance to chlorpyrifos in the offspring [92]; in the freshwater water flea Moina dubia, the antagonistic interaction between warming and lead (Pb) increased gradually across 10 exposed generations [93]. Similarly to our findings, in the planktonic copepods Pseudodiaptomus annandalei and P. incisus, parental exposure to Cu and a simulated marine heatwave resulted in lower performance of offspring [56,94]. Finally, warming interacted with nickel (Ni) and increased its multigenerational toxicity, reducing survival and reproduction of the benthic copepod T. japonicus [52].

5. Conclusions

The present study investigated the chronic toxicological effects of CBZ combined with temperature on the copepod T. fulvus over two generations. In addition to the confirmed capability of CBZ to affect development and reproduction of T. fulvus [40], the results obtained evidenced that temperature may modulate the sensitivity of such copepods to CBZ. Indeed, while at 20 °C, the exposure to CBZ inhibited the development of T. fulvus, at 27 °C, and also after CBZ exposure, copepods developed significantly faster, evidencing that the effect of the pharmaceutical was mitigated by the influence of temperature on this life trait. In contrast, growth and reproduction inhibition was greater under the warmer condition, especially in the first generation, as evidenced by co-exposed females, in which the greatest reduction in body size and fecundity was observed. The reduction in the reproductive fitness could compromise copepod population structure across generations. Although ocean warming and pharmaceutical exposure are major threats to aquatic ecosystems, and temperature can strongly alter contaminants’ toxicity, few studies have focused on how their combined effects can be transmitted to the next generation. The findings reported here highlight the need to consider maternal effects in environmental risk assessment for multiple stressors, as the influence of pollutants during single-generation exposure may be underestimated.

Author Contributions

Conceptualization, I.P., E.P. and F.B.; Methodology, F.B.; Formal analysis, G.D.; Investigation, I.P.; Data curation, I.P. and G.D.; Writing—original draft, I.P. and F.B.; Writing—review & editing, E.P.; Supervision, E.P. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors on request.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Hughes, S.R.; Kay, P.; Brown, L.E. Global synthesis and critical evaluation of pharmaceutical data sets collected from river systems. Environ. Sci. Technol. 2013, 47, 661–677. [Google Scholar] [CrossRef]
  2. Directive 2000/60/EC of the European Parliament and of the Council of 23 October 2000 Establishing a Framework for Community Action in the Field of Water Policy. Off. J. Eur. Union 2000, 1–73. Available online: http://data.europa.eu/eli/dir/2000/60/oj (accessed on 12 March 2024).
  3. Monteiro, S.C.; Boxall, A.B.A. Occurrence and Fate of Human Pharmaceuticals in the Environment. Rev. Environ. Contam. Toxicol. 2010, 202, 53–154. [Google Scholar] [CrossRef] [PubMed]
  4. Gunnarsson, L.; Jauhiainen, A.; Kristiansson, E.; Nerman, O.; Larsson, D.G.J. Evolutionary conservation of human drug targets in organisms used for environmental risk assessments. Environ. Sci. Technol. 2008, 42, 5807–5813. [Google Scholar] [CrossRef]
  5. Biel-Maeso, M.; Baena-Nogueras, R.M.; Corada-Fernández, C.; Lara-Martín, P.A. Occurrence, distribution and environmental risk of pharmaceutically active compounds (PhACs) in coastal and ocean waters from the Gulf of Cadiz (SW Spain). Sci. Total Environ. 2018, 612, 649–659. [Google Scholar] [CrossRef] [PubMed]
  6. González-Alonso, S.; Merino, L.M.; Esteban, S.; López de Alda, M.; Barceló, D.; Durán, J.J.; López-Martínez, J.; Aceña, J.; Pérez, S.; Mastroianni, N.; et al. Occurrence of pharmaceutical, recreational and psychotropic drug residues in surface water on the northern Antarctic Peninsula region. Environ. Pollut. 2017, 229, 241–254. [Google Scholar] [CrossRef]
  7. EU. COM/2019/128 Final: Communication from the Commission to the European Parliament, the Council and the European Economic and Social Committee: European Union Strategic Approach to Pharmaceuticals in the Environment. 2019. Available online: https://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX%3A52019DC0128 (accessed on 12 March 2024).
  8. Halpern, B.S.; Walbridge, S.; Selkoe, K.A.; Kappel, C.V.; Micheli, F.; D’Agrosa, C.; Bruno, J.F.; Casey, K.S.; Ebert, C.; Fox, H.E.; et al. A global map of human impact on marine ecosystems. Science 2008, 319, 948–952. [Google Scholar] [CrossRef]
  9. IPCC. Climate Change 2013: The Physical Science Basis. Contribution of the Working Group I to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change; IPCC: Cambridge, MA, USA, 2013; p. 1535. ISBN 978-92-9169-138-8. [Google Scholar]
  10. Darmaraki, S.; Somot, S.; Sevault, F.; Nabat, P.; Cabos, N.; William, D.; Cavicchia, L.; Djurdjevic, V.; Li, L.; Sannino, G.; et al. Future evolution of Marine Heatwaves in the Mediterranean Sea. Clim. Dyn. 2019, 53, 1371–1392. [Google Scholar] [CrossRef]
  11. Cabral, H.; Fonseca, V.; Sousa, T.; Costa Leal, M. Synergistic Effects of Climate Change and Marine Pollution: An Overlooked Interaction in Coastal and Estuarine Areas. Int. J. Environ. Res. Public Health 2019, 16, 2737. [Google Scholar] [CrossRef]
  12. Crain, C.M.; Kroeker, K.; Halpern, B.S. Interactive and cumulative effects of multiple human stressors in marine systems. Ecol. Lett. 2008, 11, 1304–1315. [Google Scholar] [CrossRef]
  13. Hernandez-Delgado, E.A. The emerging threats of climate change on tropical coastal ecosystem services, public health, local economies and livelihood sustainability of small islands: Cumulative impacts and synergies. Mar. Pollut. Bull. 2015, 101, 5–28. [Google Scholar] [CrossRef] [PubMed]
  14. Sokolova, I. Bioenergetics in environmental adaptation and stress tolerance of aquatic ectotherms: Linking physiology and ecology in a multi-stressor landscape. J. Exp. Biol. 2021, 224 (Suppl. S1), jeb236802. [Google Scholar] [CrossRef] [PubMed]
  15. Hemraj, D.A.; Posnett, N.C.; Minuti, J.J.; Firth, L.B.; Russell, B.D. Survived but not safe: Marine heatwave hinders metabolism in two gastropod survivors. Mar. Environ. Res. 2020, 162, 105117. [Google Scholar] [CrossRef] [PubMed]
  16. Ko, G.W.; Dineshram, R.; Campanati, C.; Chan, V.B.; Havenhand, J.; Thiyagarajan, V. Interactive effects of ocean acidification, elevated temperature, and reduced salinity on early-life stages of the pacific oyster. Environ. Sci. Technol. 2014, 48, 10079–10088. [Google Scholar] [CrossRef] [PubMed]
  17. Thiyagarajan, V.; Ko, G.W.K. Larval growth response of the Portuguese oyster (Crassostrea angulata) to multiple climate change stressors. Aquaculture 2012, 370, 90–95. [Google Scholar] [CrossRef]
  18. Noyes, P.D.; McElwee, M.K.; Miller, H.D.; Clark, B.W.; Van Tiem, L.A.; Walcott, K.C.; Erwin, K.N.; Levin, E.D. The toxicology of climate change: Environmental contaminants in a warming world. Environ. Int. 2009, 35, 971–986. [Google Scholar] [CrossRef]
  19. Alava, J.J.; Cheung, W.W.L.; Ross, P.S.; Sumaila, U.R. Climate change-contaminant interactions in marine food webs: Toward a conceptual framework. Glob. Chang. Biol. 2017, 23, 3984–4001. [Google Scholar] [CrossRef] [PubMed]
  20. Heugens, E.H.; Hendriks, A.J.; Dekker, T.; Van Straalen, N.M.; Admiraal, W. A review of the effects of multiple stressors on aquatic organisms and analysis of uncertainty factors for use in risk assessment. Crit. Rev. Toxicol. 2001, 31, 247–284. [Google Scholar] [CrossRef] [PubMed]
  21. Zhang, Y.; Geissen, S.U.; Gal, C. Carbamazepine and diclofenac: Removal in wastewater treatment plants and occurrence in water bodies. Chemosphere 2008, 73, 1151–1161. [Google Scholar] [CrossRef]
  22. Zhou, S.; Xia, Y.; Li, T.; Yao, T.; Shi, Z.; Zhu, S.; Gao, N. Degradation of carbamazepine by uv/chlorine advanced oxidation process and formation of disinfection by-products. Environ. Sci. Pollut. Res. 2016, 23, 16448–16455. [Google Scholar] [CrossRef]
  23. Zhu, S.; Dong, B.; Wu, Y.; Buc, L.; Zhou, S. Degradation of carbamazepine by vacuum-UV oxidation process: Kinetics modeling and energy efficiency. J. Hazard. Mater. 2019, 368, 178–185. [Google Scholar] [CrossRef] [PubMed]
  24. Mezzelani, M.; Fattorini, D.; Gorbi, S.; Nigro, M.; Regoli, F. Human pharmaceuticals in marine mussels: Evidence of sneaky environmental hazard along Italian coasts. Mar. Environ. Res. 2020, 162, 105137. [Google Scholar] [CrossRef]
  25. Miller, T.H.; Ng, K.T.; Bury, S.T.; Bury, S.E.; Bury, N.R.; Barron, L.P. Biomonitoring of pesticides, pharmaceuticals and illicit drugs in a freshwater invertebrate to estimate toxic or effect pressure. Environ. Int. 2019, 129, 595–606. [Google Scholar] [CrossRef]
  26. Alvarez-Munoz, D.; Rodriguez-Mozaz, S.; Maulvault, A.L.; Tediosi, A.; Fernandez-Tejedor, M.; Van den Heuvel, F.; Kotterman, M.; Marques, A.; Barcelo, D. Occurrence of pharmaceuticals and endocrine disrupting compounds in macroalgaes, bivalves, and fish from coastal areas in Europe. Environ. Res. 2015, 143, 56–64. [Google Scholar] [CrossRef] [PubMed]
  27. Kumar, A.; Batley, G.E.; Nidumolu, B.; Hutchinson, T.H. Derivation of water quality guidelines for priority pharmaceuticals. Environ. Toxicol. Chem. 2016, 35, 1815–1824. [Google Scholar] [CrossRef] [PubMed]
  28. Nardi, A.; Mezzelani, M.; Costa, S.; d’Errico, G.; Benedetti, M.; Gorbi, S.; Freitas, R.; Regoli, F. Marine heatwaves hamper neuro-immune and oxidative tolerance toward carbamazepine in Mytilus galloprovincialis. Environ. Pollut. 2022, 300, 118970. [Google Scholar] [CrossRef]
  29. Mezzelani, M.; Nardi, A.; Bernardini, I.; Mila, M.; Peruzza, L.; d’Errico, G.; Fattorini, D.; Gorbi, S.; Patarnello, T.; Regoli, F. Environmental pharmaceuticals and climate change: The case study of carbamazepine in M. galloprovincialis under ocean acidification scenario. Environ. Int. 2021, 146, 106269. [Google Scholar] [CrossRef]
  30. Costa, S.; Coppola, F.; Pretti, C.; Intorre, L.; Meucci, V.; Soares, A.M.V.M.; Freitas, R.; Solé, M. The influence of climate change related factors on the response of two clam species to diclofenac. Ecotoxicol. Environ. Saf. 2020, 189, 109899. [Google Scholar] [CrossRef]
  31. Almeida, A.; Soares, A.M.V.M.; Esteves, V.I.; Freitas, R. Occurrence of the antiepileptic carbamazepine in water and bivalves from marine environments: A review. Environ. Toxicol. Pharmacol. 2021, 86, 103661. [Google Scholar] [CrossRef]
  32. Martin-Diaz, L.; Franzellitti, S.; Buratti, S.; Valbonesi, P.; Capuzzo, A.; Fabbri, E. Effects of environmental concentrations of the antiepilectic drug carbamazepine on biomarkers and cAMP-mediated cell signaling in the mussel Mytilus galloprovincialis. Aquat. Toxicol. 2009, 94, 177–185. [Google Scholar] [CrossRef]
  33. Chen, H.; Zha, J.; Liang, X.; Li, J.; Wang, Z. Effects of the human antiepileptic drug carbamazepine on the behavior, biomarkers, and heat shock proteins in the Asian clam Corbicula fluminea. Aquat. Toxicol. 2014, 155, 1–8. [Google Scholar] [CrossRef] [PubMed]
  34. Freitas, R.; Almeida, A.; Pires, A.; Velez, C.; Calisto, V.; Schneider, R.J.; Esteves, V.I.; Wrona, F.J.; Figueira, E.; Soares, A.M. The effects of carbamazepine on macroinvertebrate species: Comparing bivalves and polychaetes biochemical responses. Water Res. 2015, 85, 137–147. [Google Scholar] [CrossRef] [PubMed]
  35. Rivetti, C.; Campos, B.; Barata, C. Low environmental levels of neuro-active pharmaceuticals alter phototactic behaviour and reproduction in Daphnia magna. Aquat. Toxicol. 2016, 170, 289–296. [Google Scholar] [CrossRef] [PubMed]
  36. Almeida, A.; Calisto, V.; Domingues, M.R.; Esteves, V.I.; Schneider, R.J.; Soares, A.M.; Figueira, E.; Freitas, R. Comparison of the toxicological impacts of carbamazepine and a mixture of its photodegradation products in Scrobicularia plana. J. Hazard. Mater. 2017, 323, 220–232. [Google Scholar] [CrossRef] [PubMed]
  37. Yan, S.; Wang, M.; Zha, J.; Zhu, L.; Li, W.; Luo, Q.; Sun, J.; Wang, Z. Environmentally relevant concentrations of carbamazepine caused endocrine-disrupting effects on nontarget organisms, Chinese rare minnows (Gobiocypris rarus). Environ. Sci. Technol. 2018, 52, 886–894. [Google Scholar] [CrossRef] [PubMed]
  38. Aguirre-Martínez, G.; Del Valls, T.; Martín-Díaz, M. Early responses measured in the brachyuran crab Carcinus maenas exposed to carbamazepine and novobiocin: Application of a 2-tier approach. Ecotoxicol. Environ. Saf. 2013, 97, 47–58. [Google Scholar] [CrossRef] [PubMed]
  39. Chen, H.; Gu, X.; Zeng, Q.; Mao, Z.; Liang, X.; Martyniuk, C.J. Carbamazepine disrupts molting hormone signaling and inhibits molting and growth of Eriocheir sinensis at environmentally relevant concentrations. Aquat. Toxicol. 2019, 208, 138–145. [Google Scholar] [CrossRef] [PubMed]
  40. Prato, E.; Biandolino, F.; Grattagliano, A.; Ruscito, A.; Lofrano, G.; Libralato, G.; Trifuoggi, M.; Albarano, L.; Parlapiano, I. Individual and combined effects of amoxicillin and carbamazepine to the marine copepod Tigriopus fulvus. Environ. Sci. Pollut. Res. 2023, 30, 61672–61681. [Google Scholar] [CrossRef]
  41. Faraponova, O.; Giacco, E.; Biandolino, F.; Prato, E.; Del Prete, F.; Valenti, A.; Sarcina, S.; Pasteris, A.; Montecavalli, A.; Comin, S.; et al. Tigriopus fulvus: The interlaboratory comparison of the acute toxicity test. Ecotoxicol. Environ. Saf. 2016, 124, 309–314. [Google Scholar] [CrossRef]
  42. Biandolino, F.; Parlapiano, I.; Faraponova, O.; Prato, E. Effects of short-and long-term exposures to copper on lethal and reproductive endpoints of the harpacticoid copepod Tigriopus fulvus. Ecotoxicol. Environ. Saf. 2018, 147, 327–333. [Google Scholar] [CrossRef]
  43. Parlapiano, I.; Biandolino, F.; Grattagliano, A.; Ruscito, A.; Libralato, G.; Prato, E. Effects of commercial formulations of glyphosate on marine crustaceans and implications for risk assessment under temperature changes. Ecotoxicol. Environ. Saf. 2021, 213, 112068. [Google Scholar] [CrossRef] [PubMed]
  44. Prato, E.; Biandolino, F.; Bisci, A.P.; Caroppo, C. Preliminary assessment of Ostreopsis cfr. ovata acute toxicity by using a battery bioassay. Chem. Ecol. 2011, 27, 117–125. [Google Scholar] [CrossRef]
  45. Prato, E.; Parlapiano, I.; Biandolino, F. Assessment of individual and combined toxicities of three metals (Cu, Cd and Hg) by using Tigriopus fulvus. Chem. Ecol. 2013, 29, 635642. [Google Scholar] [CrossRef]
  46. Prato, E.; Parlapiano, I.; Biandolino, F.; Rotini, A.; Manfra, L.; Berducci, M.T.; Maggi, C.; Libralato, G.; Paduano, L.; Carraturo, F.; et al. Chronic sublethal effects of ZnO nanoparticles on Tigriopus fulvus (Copepoda, Harpacticoida). Environ. Sci. Pollut. Res. 2020, 27, 30957–30968. [Google Scholar] [CrossRef] [PubMed]
  47. Prato, E.; Biandolino, F.; Parlapiano, I.; Grattagliano, A.; Rotolo, F.; Buttino, I. Historical control data of ecotoxicological test with the copepod Tigriopus fulvus. Chem. Ecol. 2023, 39, 881–893. [Google Scholar] [CrossRef]
  48. Bonello, G.; Angelini, C.; Pane, L. Effects of environmental factors on Tigriopus fulvus, Fischer 1860, a Mediterranean harpacticoid copepod. J. Biol. Res. 2018, 91, 7113. [Google Scholar] [CrossRef]
  49. Raisuddin, S.; Kwok, K.W.H.; Leung, K.M.Y.; Schlenk, D.; Lee, J.S. The copepod Tigriopus: A promising marine model organism for ecotoxicology and environmental genomics. Aquat. Toxicol. 2007, 83, 161–173. [Google Scholar] [CrossRef] [PubMed]
  50. Sun, P.Y.; Foley, H.B.; Handschumacher, L.; Suzuki, A.; Karamanukyan, T.; Edmands, S. Acclimation and adaptation to common marine pollutants in the copepod Tigriopus californicus. Chemosphere 2014, 112, 465–471. [Google Scholar] [CrossRef] [PubMed]
  51. Liu, T.; Zhang, L. Multigenerational effects of arsenate on development and reproduction in marine copepod Tigriopus japonicus. Chemosphere 2023, 342, 140158. [Google Scholar] [CrossRef]
  52. Zhang, Y.; Xie, D.; Lin, Q.; Zhou, X. Seawater warming intensifies nickel toxicity to a marine copepod: A multigenerational perspective. Aquat. Toxicol. 2023, 264, 106730. [Google Scholar] [CrossRef]
  53. Donelson, J.M.; Munday, P.L.; McCormick, M.I.; Pitcher, C.R. Rapid transgenerational acclimation of a tropical reef fish to climate change. Nat. Clim. Chang. 2012, 2, 30–32. [Google Scholar] [CrossRef]
  54. Kimberly, D.A.; Salice, C.J. Complex interactions between climate change and toxicants: Evidence that temperature variability increases sensitivity to cadmium. Ecotoxicology 2014, 23, 809–817. [Google Scholar] [CrossRef]
  55. Tran, T.T.; Janssens, L.; Dinh, K.V.; Stoks, R. Transgenerational interactions between pesticide exposure and warming in a vector mosquito. Evol. Appl. 2018, 11, 906–917. [Google Scholar] [CrossRef] [PubMed]
  56. Dinh, K.V.; Nguyen, Q.T.T.; Vo, T.M.; Bui, T.B.; Dao, T.S.; Tran, D.M.; Doan, N.X.; Truong, T.S.H.; Wisz, M.S.; Nielsen, T.G.; et al. Interactive effects of extreme temperature and a widespread coastal metal contaminant reduce the fitness of a common tropical copepod across generations. Mar. Pollut. Bull. 2020, 159, 111509. [Google Scholar] [CrossRef] [PubMed]
  57. Estaque, T.; Richaume, J.; Bianchimani, O.; Schull, Q.; Mérigot, B.; Bensoussan, N.; Bonhomme, P.; Vouriot, P.; Sartoretto, S.; Monfort, T.; et al. Marine heatwaves on the rise: One of the strongest ever observed mass mortality event in temperate gorgonians. Glob. Chang. Biol. 2023, 29, 6159–6162. [Google Scholar] [CrossRef] [PubMed]
  58. UNICHIM 2396: 2014. Qualità dell’acqua—Determinazione della Tossicità Letale a 24h, 48h e 96h di Esposizione con Naupli di Tigriopus fulvus (Fischer, 1860). (Crustacea:Copepoda). 2014. Available online: https://pubblicazioni.unichim.it/metodi) (accessed on 12 March 2024).
  59. Andreozzi, R.; Raffaele, M.; Nicklas, P. Pharmaceuticals in STP effluents and their solar photodegradation in aquatic environment. Chemosphere 2003, 50, 1319–1330. [Google Scholar] [CrossRef] [PubMed]
  60. Dinh, K.V.; Konestabo, H.S.; Borgå, K.; Hylland, K.; Macaulay, S.; Jackson, M.; Verheyen, J.; Stoks, R. Interactive effects of warming and pollutants on marine and freshwater invertebrates. Curr. Pollut. Rep. 2022, 8, 341–359. [Google Scholar] [CrossRef]
  61. Oropesa, A.L.; Floro, A.M.; Palma, P. Assessment of the effects of the carbamazepine on the endogenous endocrine system of Daphnia magna. Environ. Sci. Pollut. Res. 2016, 23, 17311–17321. [Google Scholar] [CrossRef] [PubMed]
  62. Chen, H.; Gu, X.; Zeng, Q.; Mao, Z. Acute and chronic toxicity of carbamazepine on the release of chitobiase, molting, and reproduction in Daphnia similis. Int. J. Environ. Res. Public Health 2019, 16, 209. [Google Scholar] [CrossRef]
  63. Nkoom, M.; Lu, G.; Liu, J. Chronic toxicity of diclofenac, carbamazepine and their mixture to Daphnia magna: A comparative two-generational study. Environ. Sci. Pollut. Res. 2022, 29, 58963–58979. [Google Scholar] [CrossRef]
  64. Nieto, E.; Hampel, M.; González-Ortegón, E.; Drake, P.; Blasco, J. Influence of temperature on toxicity of single pharmaceuticals and mixtures, in the crustacean A. desmarestii. J. Hazard. Mater. 2016, 313, 159–169. [Google Scholar] [CrossRef] [PubMed]
  65. Vernouillet, G.; Eullaffroy, P.; Lajeunesse, A.; Blaise, C.; Gagné, F.; Juneau, P. Toxic effects and bioaccumulation of carbamazepine evaluated by biomarkers measured in organisms of different trophic levels. Chemosphere 2010, 80, 1062–1068. [Google Scholar] [CrossRef] [PubMed]
  66. Garcia, S.N.; Foster, M.; Constantine, L.A.; Huggett, D.B. Field and laboratory fish tissue accumulation of the anti-convulsant drug carbamazepine. Ecotoxicol. Environ. Saf. 2012, 84, 207–211. [Google Scholar] [CrossRef]
  67. Freitas, R.; Almeida, A.; Calisto, V.; Velez, C.; Moreira, A.; Schneider, R.J.; Esteves, V.I.; Wrona, F.J.; Figueira, E.; Soares, A.M. The impacts of pharmaceutical drugs under ocean acid cation: New data on single and combined long-term effects of carbamazepine on Scrobicularia plana. Sci. Total Environ. 2016, 541, 977–985. [Google Scholar] [CrossRef] [PubMed]
  68. Zhou, R.; Lu, G.; Yan, Z.; Jiang, R.; Bao, X. A review of the influences of microplastics on toxicity and transgenerational effects of pharmaceutical and personal care products in aquatic environment. Sci. Total Environ. 2020, 732, 139222. [Google Scholar] [CrossRef] [PubMed]
  69. Lee, H.W.; Ban, S.; Ikeda, T.; Matsuishi, T. Effect of temperature on development, growth and reproduction in the marine copepod Pseudocalanus newmani at satiating food condition. J. Plankton Res. 2003, 25, 261–271. [Google Scholar] [CrossRef]
  70. Matias-Peralta, H.; Yusoff, F.M.; Shariff, M.; Arshad, A. Effects of some environmental parameters on the reproduction and development of a tropical marine harpacticoid copepod Nitocra affinis f. californica Lang. Mar. Pollut. Bull. 2005, 51, 722–728. [Google Scholar] [CrossRef] [PubMed]
  71. Karlsson, K.; Puiac, S.; Winder, M. Life-history responses to changing temperature and salinity of the Baltic Sea copepod Eurytemora affinis. Mar. Biol. 2018, 165, 30. [Google Scholar] [CrossRef]
  72. Weydmann, A.; Walczowski, W.; Carstensen, J.; Kwaśniewski, S. Warming of Subarctic waters accelerates development of a key marine zooplankton Calanus finmarchicus. Glob. Chang. Biol. 2018, 24, 172–183. [Google Scholar] [CrossRef]
  73. Li, A.J.; Lai, R.W.S.; Zhou, G.J.; Leung, P.T.Y.; Zeng, E.Y.; Leung, K.M.Y. Joint effects of temperature and copper exposure on developmental and gene-expression responses of the marine copepod Tigriopus japonicus. Ecotoxicology 2023, 32, 336–343. [Google Scholar] [CrossRef]
  74. Thistle, D.; Eckman, J.E. What is the sex ratio of harpacticoid copepods in the deep sea? Mar. Biol. 1990, 107, 443–447. [Google Scholar] [CrossRef]
  75. Shimanaga, M.; Shirayama, Y. Sex ratio and reproductive activity of benthic copepods in bathyal Sagami Bay (1430 m), central Japan. Prog. Oceanogr. 2003, 57, 97–107. [Google Scholar] [CrossRef]
  76. Shimanaga, M.; Lee, W.; Nomaki, H.; Iijima, K. Sex ratio and gut contents of the deep-sea harpacticoid Neocervinia itoi and other Cerviniids: A possibility of reduced foraging among males. J. Crustace Biol. 2009, 29, 183–191. [Google Scholar] [CrossRef]
  77. de Juan, C.; Griffell, K.; Calbet, A.; Saiz, E. Multigenerational physiological compensation and body size reduction dampen the effects of warming on copepods. Limnol. Oceanogr. 2023, 68, 1037–1047. [Google Scholar] [CrossRef]
  78. Voordouw, M.J.; Anholt, B. Environmental sex determination in a splash pool copepod. Biol. J. Linn. Soc. 2002, 76, 511–520. [Google Scholar] [CrossRef]
  79. Koch, J.; Bui, T.T.; Lundström Belleza, E.; Brinkmann, M.; Hollert, H.; Breitholtz, M. Temperature and food quantity effects on the harpacticoid copepod Nitocra spinipes: Combining in vivo bioassays with population modeling. PLoS ONE 2017, 12, e0174384. [Google Scholar] [CrossRef]
  80. Bulmer, M.G.; Bull, J.J. Models of Polygenic Sex Determination and Sex Ratio Control. Evolution 1982, 36, 13–26. [Google Scholar] [CrossRef]
  81. Horne, C.R.; Hirst, A.G.; Atkinson, D.; Neves, A.; Kiørboe, T. A global synthesis of seasonal temperature–size responses in copepods. Glob. Ecol. Biogeogr. 2016, 25, 988–999. [Google Scholar] [CrossRef]
  82. Moore, M.V.; Folt, C.F.; Stemberger, R.S. Consequences of elevated temperatures for zooplankton assemblages in temperate lakes. Arch. Hydrobiol. 1996, 135, 289–319. [Google Scholar] [CrossRef]
  83. Garzke, J.; Ismar, S.M.H.; Sommer, U. Climate change affects low trophic level marine consumers: Warming decreases copepod size and abundance. Oecologia 2015, 177, 849–860. [Google Scholar] [CrossRef]
  84. Doan, N.X.; Vu, M.T.T.; Pham, H.Q.; Wisz, M.S.; Nielsen, T.G.; Dinh, K.V. Extreme temperature impairs growth and productivity in a common tropical marine copepod. Sci. Rep. 2019, 9, 4550. [Google Scholar] [CrossRef]
  85. Rice, E.; Dam, H.G.; Stewart, G. Impact of climate change on estuarine zooplankton: Surface water warming in Long Island Sound is associated with changes in copepod size and community Structure. Estuaries Coasts 2015, 38, 13–23. [Google Scholar] [CrossRef]
  86. Kim, H.Y.; Yu, S.; Jeong, T.Y.; Kim, S.D. Relationship between trans-generational effects of tetracycline on Daphnia magna at the physiological and whole organism level. Environ. Pollut. 2014, 191, 111–118. [Google Scholar] [CrossRef]
  87. Im, H.; Na, J.; Jung, J. The effect of food availability on thermal stress in Daphnia magna: Trade-offs among oxidative stress, somatic growth, and reproduction. Aquat. Ecol. 2020, 54, 1201–1210. [Google Scholar] [CrossRef]
  88. Lürling, M.; Sargant, E.; Roessink, I. Life-history consequences for Daphnia pulex exposed to pharmaceutical carbamazepine. Environ. Toxicol. 2006, 21, 172–180. [Google Scholar] [CrossRef]
  89. Lamichhane, K.; Garcia, S.N.; Huggett, D.B.; DeAngelis, D.L.; La Point, T.W. Chronic effects of carbamazepine on life-history strategies of Ceriodaphnia dubia in three successive generations. Arch. Environ. Contam. Toxicol. 2013, 64, 427–438. [Google Scholar] [CrossRef]
  90. He, Y.; Zhang, Y.; Zhou, W.; Freitas, R.; Zhang, Y.; Zhang, Y. Combined exposure of polystyrene microplastics and carbamazepine induced transgenerational effects on the reproduction of Daphnia magna. Environ. Sci. Pollut. Res. 2023, 30, 67596–67607. [Google Scholar] [CrossRef]
  91. Schiedek, D.; Sundelin, B.; Readman, J.W.; Macdonald, R.W. Interactions between climate change and contaminants. Mar. Pollut. Bull. 2007, 54, 1845–1856. [Google Scholar] [CrossRef]
  92. Meng, S.; Delnat, V.; Stoks, R. Multigenerational effects modify the tolerance of mosquito larvaes to chlorpyrifos but not to a heat spike and do not change their synergism. Environ. Pollut. 2022, 292, 118333. [Google Scholar] [CrossRef]
  93. Pham, H.T.; Dinh, K.V.; Nguyen, C.C.; Quoc, L.B. Changes in the magnitude of the individual and combined effects of contaminants, warming, and predators on tropical cladocerans across 11 generations. Environ. Sci. Technol. 2020, 54, 15287–15295. [Google Scholar] [CrossRef]
  94. Dinh, K.V.; Doan, K.L.U.; Doan, N.X.; Pham, H.Q.; Le, T.H.O.; Le, M.H.; Vu, M.T.T.; Dahms, H.U.; Truong, K.N. Parental exposures increase the vulnerability of copepod offspring to copper and a simulated marine heatwave. Environ. Pollut. 2021, 287, 117603. [Google Scholar] [CrossRef]
Water 16 02051 g001
Figure 1. Effect of carbamazepine (CBZ) on life traits of Tigriopus fulvus in the F0 generation at 20 °C and 27 °C: (A) development time from naupliar to copepodite stage (N-C) and from naupliar to adult stage (i.e., ovigerous female) (N-Fov); (B) sex ratio (F/M); (C) body length of males and females; (D) hatching time; (E) number of nauplii per female; (F) number of broods per female; (G) number of nauplii per brood; (H) unhatched broods. Bars with different letters indicate significant differences between treatments and temperatures (p < 0.05).
Figure 1. Effect of carbamazepine (CBZ) on life traits of Tigriopus fulvus in the F0 generation at 20 °C and 27 °C: (A) development time from naupliar to copepodite stage (N-C) and from naupliar to adult stage (i.e., ovigerous female) (N-Fov); (B) sex ratio (F/M); (C) body length of males and females; (D) hatching time; (E) number of nauplii per female; (F) number of broods per female; (G) number of nauplii per brood; (H) unhatched broods. Bars with different letters indicate significant differences between treatments and temperatures (p < 0.05).
Water 16 02051 g001
Water 16 02051 g002
Figure 2. Effect of carbamazepine (CBZ) on life traits of Tigriopus fulvus in the F1 generation at 20 °C and 27 °C: (A) development time from naupliar to copepodite stage (N-C) and from naupliar to adult stage (i.e., ovigerous female) (N-Fov); (B) sex ratio (F/M); (C) body length of males and females; (D) hatching time; (E) number of nauplii per female; (F) number of broods per female; (G) number of nauplii per brood; (H) unhatched broods. Bars with different letters indicate significant differences between treatments and temperatures (p < 0.05).
Figure 2. Effect of carbamazepine (CBZ) on life traits of Tigriopus fulvus in the F1 generation at 20 °C and 27 °C: (A) development time from naupliar to copepodite stage (N-C) and from naupliar to adult stage (i.e., ovigerous female) (N-Fov); (B) sex ratio (F/M); (C) body length of males and females; (D) hatching time; (E) number of nauplii per female; (F) number of broods per female; (G) number of nauplii per brood; (H) unhatched broods. Bars with different letters indicate significant differences between treatments and temperatures (p < 0.05).
Water 16 02051 g002
Table 1. Results of the different tested endpoints (mean ± SD) for Tigriopus fulvus exposed to carbamazepine (CBZ) in the F0 and F1 generations at 20 °C and at 27 °C.
Table 1. Results of the different tested endpoints (mean ± SD) for Tigriopus fulvus exposed to carbamazepine (CBZ) in the F0 and F1 generations at 20 °C and at 27 °C.
Endpoint20 °C
GenerationF0F1
CBZ (μg L−1)010 10
N-C * (days)5.46 a ± 0.565.66 b ± 0.285.46 a ± 0.565.66 c ± 0.28
N-Fov * (days)14.11 a ± 0.6814.50 a ± 0.7114.56 a± 0.5115.39 b ± 0.71
Sex ratio (F/M)1.37 ± 0.271.41 ± 0.281.11 ± 1.181.16 ± 0.16
Female length (μm)823.65 ± 90.33889.83 ± 64.35870.69 ± 113.02891.82 ± 43.74
Male length (μm)843.39 ± 97.29807.10 ± 80.00853.80 ± 74.53831.04 ± 49.10
Hatching time (days)2.49 a ± 0.232.77 b ± 0.422.52 a± 0.212.76 b ± 0.23
Nauplii/Fov117.22 ± 21.63109.11 ± 23.92107.06 ± 26.05101.83 ± 26.91
Nauplii/brood21.78 ± 3.4522.62 ± 4.2421.30 ± 3.3522.91 ± 5.29
Brood/Fov5.39 a ± 0.614.83 b,c ± 0.625.00 a,b ± 0.694.44 c ± 0.51
Unhatched broods0.06 ± 0.230.06 ± 0.230.00 ± 0.000.22 ± 0.43
27 °C
GenerationF0 F1
CBZ (μg L−1)010010
N-C * (days)4.16 a ± 0.374.14 a ± 0.374.35 b ± 0.374.40 b ± 0.37
N-Fov * (days)10.94 ± 1.3510.78 ± 1.5210.83 ± 0.9211.33 ± 0.84
Sex ratio (F/M)2.23 ± 0.763.48 ± 0.862.31 ± 0.522.65 ± 0.44
Female length (μm)859.43 a ± 67.00876.83 a ± 30.53814.73 a ± 91.61698.04 b ± 25.02
Male length (μm)795.13 ± 102.55810.97 ± 108.99848.40 ± 71.40798.53 ± 51.47
Hatching time (days)2.41 a ± 0.262.70 b ± 0.202.54 a ± 0.233.03 b ± 0.39
Nauplii/Fov *119.33 a ± 38.2786.44 b ± 37.85126.28 a ± 36.8278.78 b ± 33.94
Nauplii/brood19.51 ± 5.6015.76 ± 6.6119.31 ± 5.4616.10 ± 6.00
Brood/Fov *6.11 a ± 0.765.56 b ± 0.706.56 a ± 0.704.94 b ± 0.94
Unhatched broods 0.22 a ± 0.431.00 b ± 0.770.11 a ± 0.320.67 a ± 0.48
Notes: * Fov: ovigerous female; N-C: development time from naupliar to copepodite stage; N-Fov: development time from naupliar to adult stage (i.e., ovigerous female). Different letters indicate significant differences between both treatments and generations (p < 0.05).
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Parlapiano, I.; Prato, E.; Denti, G.; Biandolino, F. Influence of Temperature on the Toxic Effects of Carbamazepine on the Copepod Tigriopus fulvus: A Transgenerational Full Life Cycle Study. Water 2024, 16, 2051. https://doi.org/10.3390/w16142051

AMA Style

Parlapiano I, Prato E, Denti G, Biandolino F. Influence of Temperature on the Toxic Effects of Carbamazepine on the Copepod Tigriopus fulvus: A Transgenerational Full Life Cycle Study. Water. 2024; 16(14):2051. https://doi.org/10.3390/w16142051

Chicago/Turabian Style

Parlapiano, Isabella, Ermelinda Prato, Giuseppe Denti, and Francesca Biandolino. 2024. "Influence of Temperature on the Toxic Effects of Carbamazepine on the Copepod Tigriopus fulvus: A Transgenerational Full Life Cycle Study" Water 16, no. 14: 2051. https://doi.org/10.3390/w16142051

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop