Multilevel assessment of carbamazepine effects: An integrative approach using zebrafish early-life stages.

Carbamazepine (CBZ) is the most commonly used drug in epilepsy treatment, and its metabolites are commonly detected among persistent pharmaceuticals in the aquatic environment. This study aimed to investigate CBZ effects on early-life-stage zebrafish (Danio rerio) (from 2 to 168 hpf) by employing of an integrative approach linking endpoints from molecular to individual level: (i) development; (ii) locomotor activity; (iii) biochemical markers (lactate dehydrogenase, glutathione-S-transferase, acetylcholinesterase and catalase) and (iv) transcriptome analysis using microarray. A 168 h - LC50 of 73.4 mg L-1 and a 72 h - EC50 of 66.8 mg L-1 for hatching were calculated while developmental effects (oedemas and tail deformities) were observed at CBZ concentrations above 37.3 mg L-1. At the biochemical level, AChE activity proved to be the most sensitive parameter, as evidenced by its decrease across all concentrations tested (∼25% maximum reduction, LOEC (lowest observed effect concentration) < 0.6 µg L-1). Locomotor behaviour seemed to be depressed by CBZ although this effect was only evident at the highest concentration tested (50 mg L-1). Molecular analysis revealed a dose-dependent effect of CBZ on gene expression. Although only 25 genes were deregulated in organisms exposed to CBZ when compared to controls, both 0.6 and 2812 µg L-1 treatments impaired gene expression related to development (e.g. crygmxl1, org, klf2a, otos, stx16 and tob2) and the nervous system (e.g. Rtn3, Gdf10, Rtn3), while activated genes were associated with behavioural response (e.g. prlbr and taar). Altogether, our results indicate that environmentally relevant CBZ concentrations might affect biochemical and genetic traits of fish. Thus, the environmental risk of CBZ cannot be neglected, especially in a realistic scenario of constant input of domestic effluents into aquatic systems.

The Vocal Repertoire of the Bearded Capuchin (Cebidae: Sapajus libidinosus): Implications for Understanding the Complexity of Neotropical Primate Communication.

Vocal communication is an essential aspect of primate social behaviour. The bearded capuchin Sapajus libidinosus is endemic to Brazil, and some studies have described specific vocalisation types for this species; however, there is still no complete description of its vocal repertoire. Thus, this study aimed to describe the vocal repertoire of a group of S. libidinosus living in theParque Nacional de Brasília, a protected area in the Cerrado area of Central Brazil. We carried out focal samplings and recording of vocalisations of members of an S. libidinosus troop in different behavioural contexts. The call analyses revealed 25 different types of vocalisations, and each call presented significant structural variation. We grouped these vocalisations according to the context of the emission or acoustic structure into the following categories: contact calls (contact note, infant babbling, trill, teeth- and lip-smacking, and sirena); foraging calls (chihui, grgr, and patinado); whistle series (food-associated, long-distance, and intergroup encounter); aggressive calls (aggressive contact note, ascending rapid staccato, cough cough, and pip); calls in response to aggression (scream, squeal, and pulsed scream), sexual display calls (chuck and raspy oestrous call), and stress-related calls (alarm call/bark, hiccup, hip, double hip, and wah wah). S. libidinosus presented a very rich vocal repertoire, revealing a pattern consistent with the repertoire of other capuchin monkey species. This is the first comprehensive description of the S. libidinosus vocal repertoire and highlights the complexity of neotropical primate communication.

Chronic effects of carbamazepine on zebrafish: Behavioral, reproductive and biochemical endpoints.

Carbamazepine (Cbz), one of the most prescribed pharmaceuticals in the world is often detected in surface waters and sediments. However, few studies addressed its chronic effects in fish. In the present study, Danio rerio adults were exposed for 63 days to Cbz (0 - control, 10 µg L-1 - concentration found in effluents, and 10,000 µg L-1 - 5% of LC50 at 72 h). Assessed endpoints were: feeding behavior, growth rate, number of eggs produced and their viability, histological alterations in female gonads, and biochemical biomarkers associated with antioxidant defenses (catalase - CAT, and glutathione S-transferase - GST activities), neurotransmission (acetylcholinesterase activity - AChE) and metabolism (lactate dehydrogenase - LDH). Cbz exposure increased the total time for food intake but did not affect D. rerio growth. Although the total number of eggs was not affected by Cbz exposure, the eggs viability was significantly impaired. Exposure to Cbz caused alterations in the female gonads follicular stages. In terms of biochemical endpoints, CAT activity in liver and gills, was sensitive to the pharmaceutical exposure presenting a decreased activity. AChE activity was induced in the head (both concentrations) and muscle (10,000 µg L-1). GST activity was increased in gills (both concentrations) but inhibited in the intestine. Concerning LDH, enzymatic activity was increased in the liver and decreased in muscle and gills. Several of the above-mentioned effects can be directly linked with effects at population level (e.g. feeding behavior) and occurred at environmental concentrations (the lowest concentration tested), thus serious concerns regarding risks posed by Cbz residues to fish populations arise with this study.