Fish mortality in the Amazonian drought of 2023: the role of experimental biology in our response to climate change.

Higher temperatures exacerbate drought conditions by increasing evaporation rates, reducing soil moisture and altering precipitation patterns. As global temperatures rise as a result of climate change, these effects intensify, leading to more frequent and severe droughts. This link between higher temperatures and drought is particularly evident in sensitive ecosystems like the Amazon rainforest, where reduced rainfall and higher evaporation rates result in significantly lower water levels, threatening biodiversity and human livelihoods. As an example, the serious drought experienced in the Amazon basin in 2023 resulted in a significant decline in fish populations. Elevated water temperatures, reaching up to 38°C, led to mass mortality events, because these temperatures surpass the thermal tolerance of many Amazonian fish species. We know this because our group has collected data on critical thermal maxima (CTmax) for various fish species over multiple years. Additionally, warmer waters can cause hypoxia, further exacerbating fish mortality. Thus, even Amazon fish species, which have relatively high thermal tolerance, are being impacted by climate change. The Amazon drought experienced in 2023 underscores the urgent need for climate action to mitigate the devastating effects on Amazonian biodiversity. The fact that we have been able to link fish mortality events to data on the thermal tolerance of fishes emphasizes the important role of experimental biology in elucidating the mechanisms behind these events, a link that we aim to highlight in this Perspective.

Oil spill in an amazon blackwater environment: Biochemical and physiological responses of local fish species.

The accidental spill of petroleum asphalt cement (PAC) in São Raimundo (SR Harbor, located on the Rio Negro (Manaus, Amazonas, Brazil) was monitored through the analysis of polyciclic aromatic hydrocarbons (PAHs) in water and a set of biomarkers in fishes (exposure biomarkes: PAHs-type metabolites concentrations in bile; the activities of ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in liver. Effect biomarkers: lipid peroxidation concentration (LPO) in liver, acetylcholinesterase activity in brain, and genotoxic DNA damage in erythrocytes). Two fish species, Acarichthys heckelii and Satanoperca jurupari, were collected 10, 45, and 90 days after the PAC spill in São Raimundo. At the same time, fish were collected from the Tupé Sustainable Development Reserve (Tupé) which served as a reference area. The sampling periods were related to the rising waters of the natural flood pulse of the Rio Negro. Higher concentrations of PAHs in water were observed at 10 and 45 days and returned to the values of TP 90 days after the PAC spill, a period in which harbor waters rose about 0.2 m. Unlike the PAHs in water, biomarker responses in both fish species significantly increased following the PAC spill in SR. Hepatic ethoxyresorufin-O-deethylase (EROD), PAH-like metabolites in bile, and erythrocyte DNA damage increases, together with inhibition of acetylcholinesterase (AChE) activity in the brain were the most evident responses for both fish species. The calculated pyrolytic index showed mixed sources of PAHs (petrogenic and pyrolytic). The applied PCA-FA indicated important relationships between dissolved organic carbon (DOC) and PAHs concentrations in water, where DOC and PAHs concentrations contributed to biomarkers responses for both fish species in all collection periods.

Amazon fish bacterial communities show structural convergence along widespread hydrochemical gradients.

The world's richest freshwater fish community thrives in gradients of contrasting environments in Amazonia, ranging from ion-poor acidic black waters, to ion-rich circumneutral white waters. These hydrochemical gradients structure Amazonian fish assemblages via ecological speciation events. Fish bacterial communities contain an important genetic heritage essential for their hosts' survival and are also involved in adaptive divergence via niche adaptation processes, but the extent to which they evolve in response to hydrochemical gradients in Amazonia is unknown. Here we investigated bacterial communities (gut and skin mucus) of two ecologically and phylogenetically divergent host species (Mesonauta festivus and Serrasalmus rhombeus) distributed throughout these hydrochemical gradients. The goal was to characterize intra- and interspecific Amazonian fish microbiome variations across multiple scales. Using a 16S metabarcoding approach, we investigated the microbiota of 43 wild M. festivus, 32 S. rhombeus and seven water samples, collected at seven sampling sites encompassing both water colours. Taxonomical structures of bacterial communities from both host species were significantly correlated to the environmental continua of magnesium, sodium, dissolved organic carbon, calcium, dissolved O2 , pH, potassium, hardness and chloride. Analysis of discriminating features in community structures across multiple scales demonstrated intra- and interspecific structural parallelisms in the response to the hydrochemical gradients. Together, these parallelisms suggest the action of selection on bacterial community structures along Amazonian hydrochemical gradients. Functional approaches along with reciprocal transplant experiments will provide further insights on the potential contribution of Amazonian fish microbiomes in host adaptation and ecological speciation events.

The use of clove oil as an anesthetic for advanced juvenile tilapia (Oreochromis niloticus) - doi: 10.4025/actascianimsci.v34i2.13022 / O uso do óleo de cravo como anestésico em juvenis avançados de tilápia do Nilo (Oreochromis niloticus) - doi: 10.4025/actascianimsci.v34i2.13022

Handling of live fish is inevitable in fish farms and causes a number of adverse physiological reactions, thus requiring the use of anesthetics. The main goal of the present work was to evaluate the efficiency of clove oil as an anesthetic for juvenile tilapia (approximately 55 g and 13 cm) during handling. In the first experiment, fishes were exposed to an anesthetic bath at six concentrations (80, 100, 150, 200, 250 and 300 mg L-1). Subsequently, the induction time was assessed for different stages of anesthesia. In the second experiment, different times of exposure to anesthesia were tested, aiming to assess the safety margin of the anesthetic. Finally, we evaluated the responses to stress after exposure to 250 mg L-1 anesthesia. The following parameters were tested: cortisol, glucose, hematocrit, hemoglobin, and plasma sodium and potassium levels. The concentration of 250 mg L-1 of clove oil is the most appropriate to induce surgical anesthesia, and 150 mg L-1 for biometrics and brief handling. Anesthesia at the ideal concentration for 10 minutes did not result in severe stress. Only blood glucose and hematocrit were affected after anesthesia.

A inevitável manipulação de peixes vivos em piscicultura causa uma série de reações fisiológicas adversas, o que exige o uso de anestésicos. O objetivo foi avaliar a eficiência do óleo de cravo como anestésico para juvenis avançados (aproximadamente 55 g e 13 cm) de tilápia do Nilo durante o manejo. No primeiro experimento, os peixes foram expostos a banhos anestésicos em seis diferentes concentrações (80; 100; 150; 200; 250 e 300 mg L-1) e foi avaliado o tempo de indução aos diferentes estágios de anestesia. No segundo experimento, avaliaram-se os diferentes tempos de exposição à anestesia, sendo verificada a margem de segurança do anestésico. Por último, avaliaram-se as respostas de estresse após a anestesia em 250 mg L-1. Os parâmetros avaliados foram cortisol, glicose, hematócrito, hemoglobina e níveis plasmáticos de sódio e potássio. A concentração 250 mg L-1 de óleo de cravo foi a mais adequada para indução de anestesia cirúrgica. Para a anestesia voltada para biometria e breve manejo, a concentração mais adequada foi 150 mg L-1. A exposição à concentração ideal de óleo de cravo por 10 min., não causou estresse severo, ocorrendo alterações após a anestesia somente nos níveis de glicose e hematócrito.

The use of clove oil as an anesthetic for advanced juvenile tilapia (Oreochromis niloticus) - doi: 10.4025/actascianimsci.v34i2.13022 / O uso do óleo de cravo como anestésico em juvenis avançados de tilápia do Nilo (Oreochromis niloticus) - doi: 10.4025/actascianimsci.v34i2.13022

Handling of live fish is inevitable in fish farms and causes a number of adverse physiological reactions, thus requiring the use of anesthetics. The main goal of the present work was to evaluate the efficiency of clove oil as an anesthetic for juvenile tilapia (approximately 55 g and 13 cm) during handling. In the first experiment, fishes were exposed to an anesthetic bath at six concentrations (80, 100, 150, 200, 250 and 300 mg L-1). Subsequently, the induction time was assessed for different stages of anesthesia. In the second experiment, different times of exposure to anesthesia were tested, aiming to assess the safety margin of the anesthetic. Finally, we evaluated the responses to stress after exposure to 250 mg L-1 anesthesia. The following parameters were tested: cortisol, glucose, hematocrit, hemoglobin, and plasma sodium and potassium levels. The concentration of 250 mg L-1 of clove oil is the most appropriate to induce surgical anesthesia, and 150 mg L-1 for biometrics and brief handling. Anesthesia at the ideal concentration for 10 minutes did not result in severe stress. Only blood glucose and hematocrit were affected after anesthesia.

A inevitável manipulação de peixes vivos em piscicultura causa uma série de reações fisiológicas adversas, o que exige o uso de anestésicos. O objetivo foi avaliar a eficiência do óleo de cravo como anestésico para juvenis avançados (aproximadamente 55 g e 13 cm) de tilápia do Nilo durante o manejo. No primeiro experimento, os peixes foram expostos a banhos anestésicos em seis diferentes concentrações (80; 100; 150; 200; 250 e 300 mg L-1) e foi avaliado o tempo de indução aos diferentes estágios de anestesia. No segundo experimento, avaliaram-se os diferentes tempos de exposição à anestesia, sendo verificada a margem de segurança do anestésico. Por último, avaliaram-se as respostas de estresse após a anestesia em 250 mg L-1. Os parâmetros avaliados foram cortisol, glicose, hematócrito, hemoglobina e níveis plasmáticos de sódio e potássio. A concentração 250 mg L-1 de óleo de cravo foi a mais adequada para indução de anestesia cirúrgica. Para a anestesia voltada para biometria e breve manejo, a concentração mais adequada foi 150 mg L-1. A exposição à concentração ideal de óleo de cravo por 10 min., não causou estresse severo, ocorrendo alterações após a anestesia somente nos níveis de glicose e hematócrito.

Velocidade crítica de natação (Ucrit) de matrinxã (Brycon amazonicus) após exposição à hipoxia

Escape is the first response of fish to stress, that depends on the swimming performance and the physiological adjustments. This study has investigated the critical swimming speed (Ucrit) of matrinxã after exposure to hypoxia. To achieve that, the fishes were exposed to hypoxia and analyzed before and after forced swimming, using Ucrit. The hypoxia caused an increase of lactate, glucose, cortisol and hematocrit. No changes of plasma sodium and potassium levels, as well as the Ucrit, were observed. We suggest that matrinxã is sensitive to hypoxia, but the physiological adjustments are sufficient to keep its swimming performance.

A primeira resposta ao estresse é a fuga, que depende do desempenho natatório e de ajustes fisiológicos. Este estudo investigou a velocidade crítica de natação (Ucrit) de matrinxã após exposição à hipoxia. Para isso, os peixes foram expostos à hipoxia, sendo uma parte do grupo analisada antes e outra após natação forçada, por meio da Ucrit. A hipoxia resultou no aumento de lactato, glicose, cortisol e hematócrito. Mudanças nos níveis de sódio e potássio, bem como os valores de Ucrit não foram observadas. Sugere-se que o matrinxã seja sensível à hipoxia, mas os ajustes fisiológicos são suficientes para manter seu desempenho natatório.

Tambaqui growth and survival when exposed to different photoperiods

The use of different photoperiods (light) were investigated during tambaqui (Colossoma macropomum) juvenile growth under captivity. Light intensity tested was: continuous dark (24hrs without light), natural photoperiod simulation (10hrs of light and 14hrs without light) and continuous light (24 with light). No mortality was recorded among treatments. Significant differences was observed after 50 days of experiment among mean fish weight, fish kept under a continuous darkness showed a better specific growth rate (6.02%) when compared to control fish (natural photo period, 3.67%). Fish exposed to continuous light presented the lowest mean specific growth rate (2.04%). It is possible to improve tambaqui juvenile weight gain performance when kept under continuous darkness.

Investigou-se o uso de diferentes fotos períodos (iluminação) durante o crescimento de juvenis de tambaqui (Colossoma macropomum) em cativeiro. As intensidades testadas foram: escuro contínuo (24hs sem luz), simulação do foto período natural (10hs de luz e 14hs sem luz) e iluminação contínua (24hs de luz). Não houve mortalidade nos diferentes tratamentos. Diferença significativa entre as médias do peso foi observada após 50 dias de experimento, a exposição ao escuro contínuo apresentou o melhor índice de crescimento específico (6,02%) em relação ao controle (foto período natural, com 3,67%). O menor índice de crescimento médio foi observado na exposição à iluminação contínua (2,04%). Assim, pode-se melhorar o desempenho de ganho de peso para juvenis de tambaqui quando mantidos no escuro contínuo.

Protection by natural blackwater against disturbances in ion fluxes caused by low pH exposure in freshwater stingrays endemic to the Rio Negro.

Stenohaline freshwater stingrays (Potamotrygon spp.) are endemic to the very dilute (Na(+), Cl(-), Ca2(+) 300 micromol L(-1) in reference water (low DOC) to about 100 micromol L(-1) in blackwater (high DOC). In reference water, both JNain and JClin were inhibited >90%, both JNaout and JClout more than doubled, and J(Amm) did not change at pH 4.0. In blackwater, the inhibition of influxes was attenuated, the increases in outflux did not occur, and J(Amm) increased by 60% at pH 4.0. Addition of 100 micromol L(-1) Ca(2+) to reference water prevented the increases in JNaout and JClout and allowed J(Amm) to increase at pH 4.0, which demonstrates that the gills are sensitive to Ca(2+). However, addition of Ca(2+) to blackwater had no effect on the responses to pH 4.0. Addition of commercial humic acid to reference water did not duplicate the effects of natural Rio Negro blackwater at the same DOC level; instead, it greatly exacerbated the increases in JNaout and JClout at low pH and prevented any protective influence of added Ca(2+). Thus, blackwater DOC appears to be very different from commercial humic acid. Biogeochemical modeling indicated that blackwater DOC prevents Ca(2+) binding, but not H(+) binding, to the gills and that the protective effects of blackwater cannot be attributed to its higher buffer capacity or its elevated Al or Fe levels. Natural DOC may act directly at the gills at low pH to exert a protective effect and, when doing so, may override any protective action of Ca(2+) that might otherwise occur.

Safest level of tricaine methanesulfonate (ms-222) to induce anesthesia in juveniles of Matrinxã, Brycon cephalus

The use of MS-222 as an anesthetic for matrinxã juveniles was investigated. At dosage of 100 mg/L or lower fish did not achieve a complete anesthesia state. At 150 mg/L, MS-222 induced anesthesia within 36 seconds and recovered from a 10 minutes period of anesthesia within 5.2 min. Higher concentrations (200, 250 and 300 mg/L) anesthetized fish in lesser times, with the offset of mortality (16.7 and 33.3%) at the 200 and 300 mg/L MS-222 doses, respectively. The only significant differences observed in the hematological parameters, was for the glucose values in fish anesthetized with 250 and 300 mg/L. From the results, the recommended dose of MS-222 for handling matrinxã juveniles is 150 mg/L.

Investigou-se o uso de MS-222 como anestésico para juvenis de matrinxã. Concentrações de 100 mg/L ou menores não causam uma indução completa à anestesia. A 150 mg/L os peixes foram induzidos à anestesia após 36 segundos e se recuperam em 5,2 minutos após exposição ao anestésico durante 10 minutos. Em concentrações maiores (200, 250 e 300 mg/L) os peixes foram induzidos a anestesia em menor tempo, porém observou-se mortalidade de 16,7 e 33,3 % nas doses de 200 e 300 mg/L, respectivamente. Nos parâmetros hematológicos observou-se uma alteração significante nos níveis de glicose para os peixes anestesiados com doses de 250 e 300 mg/L. Os resultados obtidos indicam que 150 mg/L é a dose ideal de MS-222 para anestesiar juvenis de matrinxã.

Safest level of tricaine methanesulfonate (ms-222) to induce anesthesia in juveniles of Matrinxã, Brycon cephalus

The use of MS-222 as an anesthetic for matrinxã juveniles was investigated. At dosage of 100 mg/L or lower fish did not achieve a complete anesthesia state. At 150 mg/L, MS-222 induced anesthesia within 36 seconds and recovered from a 10 minutes period of anesthesia within 5.2 min. Higher concentrations (200, 250 and 300 mg/L) anesthetized fish in lesser times, with the offset of mortality (16.7 and 33.3%) at the 200 and 300 mg/L MS-222 doses, respectively. The only significant differences observed in the hematological parameters, was for the glucose values in fish anesthetized with 250 and 300 mg/L. From the results, the recommended dose of MS-222 for handling matrinxã juveniles is 150 mg/L.

Investigou-se o uso de MS-222 como anestésico para juvenis de matrinxã. Concentrações de 100 mg/L ou menores não causam uma indução completa à anestesia. A 150 mg/L os peixes foram induzidos à anestesia após 36 segundos e se recuperam em 5,2 minutos após exposição ao anestésico durante 10 minutos. Em concentrações maiores (200, 250 e 300 mg/L) os peixes foram induzidos a anestesia em menor tempo, porém observou-se mortalidade de 16,7 e 33,3 % nas doses de 200 e 300 mg/L, respectivamente. Nos parâmetros hematológicos observou-se uma alteração significante nos níveis de glicose para os peixes anestesiados com doses de 250 e 300 mg/L. Os resultados obtidos indicam que 150 mg/L é a dose ideal de MS-222 para anestesiar juvenis de matrinxã.

Aspectos biológicos de peixes amazônicos. IV. Padrões eletroforéticos de hemoglobinas de 22 espécies coletadas na ilha da Marchantaria (Manaus - AM).

The hemoglobin patterns of hemolysates from 22 fish species belonging to Manchantaria island (Solimões river , Manaus, AM) had been characterized by starch gel and agarstarch gel electrophresis. High interespecific hemoglobin heterogeneity was observed, similarly to those described for sub-tropical and temperate fish species. The resolution power of the electrophoretic supports was considered. The adaptability question of the multiple hemoglobin systems was regarded.

Foram estudados hemolisados de 22 espécies de peixes coletados na Ilha da Marchantaria (rio Solimões, Manaus - AM), através de eletroforeses em gel de amido e gel de ágar-amido. Uma grande heterogeneidade hemoglobínica interespecífica foi detectada, a exemplo do observado para espécies de zona subtropical e temperada. A capacidade de resolução de dois suportes eletroforéticos é discutida. Os resultados são discutidos em função da possível adaptabilidade conferida pelos sistemas de múltiplas hemoglobinas.

Aspectos biológicos de peixes amazônicos. IV. Padrões eletroforéticos de hemoglobinas de 22 espécies coletadas na ilha da Marchantaria (Manaus - AM)

The hemoglobin patterns of hemolysates from 22 fish species belonging to Manchantaria island (Solimões river , Manaus, AM) had been characterized by starch gel and agarstarch gel electrophresis. High interespecific hemoglobin heterogeneity was observed, similarly to those described for sub-tropical and temperate fish species. The resolution power of the electrophoretic supports was considered. The adaptability question of the multiple hemoglobin systems was regarded.

Foram estudados hemolisados de 22 espécies de peixes coletados na Ilha da Marchantaria (rio Solimões, Manaus - AM), através de eletroforeses em gel de amido e gel de ágar-amido. Uma grande heterogeneidade hemoglobínica interespecífica foi detectada, a exemplo do observado para espécies de zona subtropical e temperada. A capacidade de resolução de dois suportes eletroforéticos é discutida. Os resultados são discutidos em função da possível adaptabilidade conferida pelos sistemas de múltiplas hemoglobinas.