Description of the life cycle of Dolops discoidalis (Bouvier, 1899) (Branchiura: Argulidae), a parasite of the fish species Rhytiodus argenteofuscus (Kner, 1858) from the Brazilian Amazon.

The genus Dolops (Audouin, 1837) is endemic among ectoparasites present in fish, with nine species described in Brazil, five of which are from the Amazon region. Among the genus, the species Dolops discoidalis (Bouvier, 1899) stands out due to its high prevalence of parasitism in Amazonian fish. The present study aims to characterize the development phases of D. discoidalis. The parasites were obtained from infected fish species Rhytiodus argenteofuscus from a natural environment, kept in polyethylene experimental boxes (500 liters). The physical-chemical parameters of the water (pH, electrical conductivity, dissolved oxygen and temperature) were monitored daily. A total of 30 egg layings were analyzed. After identification of the oviposition, the eggs were transferred to another container containing water (0.5 liters). The number of eggs was recorded, with observations made in stereomicroscopic and photographic records until hatching occurred. The egg layings exhibited differences in colour throughout their development. On the seventh day, the embryos exhibited visible ocelli and on the 17th day the hatching period began. Hatching occurred in batches, with around 5­10 hatches/day/laying. However, peak hatching was recorded on the 19th day of the cycle. Therefore, the cycle of D. discoidalis comprised a total period of 17­22 days at 28.C, with hatching in batches and pigmentation of the eggs observed close to hatching. Newly hatched parasites did not survive for more than 24 hours in the absence of the host. This study contributes to knowledge of the biology of D. discoidalis, and adds to information regarding possible studies into its control.

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae).

This study investigated the acute toxicity (LC50-24 hr ) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC50-24 hr ) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 µg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC50-24 hr for Argulus sp. adults was 67.97 µg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 µg/L, while the LC50-24 hr for Argulus sp. and D. discoidalis was 83.98 µg/L and 82.48 µg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.

Citral chemotype of the Lippia alba essential oil as an additive in simulated transport with different loading densities of tambaqui juveniles / Óleo essencial de Lippia alba quimiotipo citral como aditivo em transporte simulado de juvenis de Tambaqui com diferentes densidades

ABSTRACT: The objective of this study was to evaluate the efficacy of the citral chemotype of Lippia alba essential oil (EOLA) as an additive in a simulated transport with different loading densities of tambaqui (Colossoma macropomum) juveniles. Simulated transport of tambaqui juveniles at three loading densities (30, 60 and 90 fish/L) was carried out either with only water (WC) or 20 μL.L-1 EOLA. Dissolved oxygen levels, pH, conductivity and temperature of the water were measured at the beginning and end of the simulated transport. Treatment with EOLA did not change survival or dissolved oxygen levels at the end of simulation. The pH of the water at the end of simulation with EOLA was lower than in that of WC but did not change with loading density. Water conductivity increased with loading density; however, it was lower in the groups with EOLA. The addition of EOLA can be useful in long-term transportation at high loading densities, but further studies related to tambaqui physiology are required.

RESUMO: O objetivo deste estudo foi avaliar a eficácia do óleo essencial de Lippia alba quimiotipo citral (EOLA) como aditivo em transporte simulado com diferentes densidades de carga de juvenis de tambaqui (Colossoma macropomum). O transporte simulado de juvenis de tambaqui em três densidades de carga (30, 60 e 90 peixes / L) foi realizado apenas com água (WC) ou 20 μL.L-1 EOLA. Níveis de oxigênio dissolvido, pH, condutividade e temperatura da água foram medidos no início e no final do transporte simulado. O tratamento com EOLA não alterou a sobrevivência ou os níveis de oxigênio dissolvido no final da simulação. O pH da água no final da simulação com o EOLA foi menor que o do WC, mas não mudou com a densidade de carga. A condutividade da água aumentou com a densidade de carga, no entanto foi menor nos grupos com EOLA. A adição de EOLA pode ser útil no transporte de longo prazo em altas densidades de carga, mas são necessários mais estudos relacionados à fisiologia do tambaqui.

Anesthetic induction of juveniles of Rhamdia quelen and Ctenopharyngodon idella with Ocimum micranthum essential oil / Indução anestésica de Rhamdia quelen e Ctenopharyngodon idella com o óleo essencial de Ocimum micranthum

ABSTRACT: Anesthetics are effective to reduce or minimize stress in handling operations related to fish cultures. Aiming to avoid residual effects of synthetic drugs, several natural agents, such as essential oils (EOs), have been tested. The aim of this study was to determine the optimal concentration of the EO of Ocimum micranthum for induction of anesthesia in juveniles of silver catfish (Rhamdia quelen) and grass carp (Ctenopharyngodon idella). Results demonstrated that the most suitable concentration to sedate silver catfish and carp is 25μL/L, while to anesthetize 200μL/L is more suitable for silver catfish and 100μL/L for grass carp, because these concentrations induce deep anesthesia in less than three min and recovery in less than five min. Analysis of the EO constituents showed methyl chavicol, a compound with carcinogenic potential, as the major component (58.2%). Therefore, the EO of O. micranthum is not recommended for fish anesthesia if the fish are to be used as food.

RESUMO: Os anestésicos são eficazes para reduzir ou minimizar o estresse nas operações de manuseio relacionadas à piscicultura. Com o objetivo de evitar efeitos residuais de fármacos sintéticos, vários agentes naturais, como óleos essenciais (OEs), têm sido testados. O objetivo deste estudo foi determinar a concentração ótima do OE de Ocimum micranthum para a indução de anestesia em juvenis de jundiá (Rhamdia quelen) e carpa capim (Ctenopharyngodon idella). Os resultados demonstram que a concentração mais adequada para sedar jundiás e carpas capim é 25μL/L. Enquanto que para anestesiar 200μL/L é mais adequado para jundiás e 100μL/L para carpas capim, pois essas concentrações induzem anestesia profunda em menos de três minutos e recuperação em menos de cinco minutos. A análise dos constituintes do OE identificou o metil chavicol, um composto com potencial carcinogênico, como componente majoritário (58,2%). Portanto, o OE de O. micranthum não é recomendado para anestesia de peixes usado como alimento.

Anesthetic potential of the essential oils of Lippia alba and Lippia origanoides in Tambaqui juveniles / Potencial anestésico do óleo essencial de Lippia Alba e Lippia origanoides em juvenis de Tambaqui

ABSTRACT: The aim of this study was to investigate the anesthetic effect of the essential oils (EOs) of Lippia alba (citral and linalool chemotypes, EOLA-C and EOLA-L respectively) and Lippia origanoides (chemotype carvacrol, EOLO-Ca) on tambaqui (Colossoma macropomum) juveniles. The tested concentrations of the L. alba and L. origanoides EOs were: 25, 50, 100, and 200 μL/L. Time required to reach the stages of sedation, deep anesthesia, and recovery, as well as ventilatory frequency, were monitored for each EO concentration. The results indicated that all EO concentrations tested induced tambaqui anesthesia and that the ventilatory frequency was altered by the EOs. L. origanoides showed the shortest time to induce sedation and anesthesia, but had long recovery times. L. alba EO (citral 100 μL/L and linalool 200 μL/L) can be used to efficiently anaesthetise tambaqui juveniles with an efficient recovery.

RESUMO: O objetivo deste estudo foi investigar o efeito anestésico dos óleos essenciais (EOs) de Lippia alba (quimiotipos citral e linalool - EOLA-C e EOLA-L respectivamente) e Lippia origanoides (quimiotipo carvacrol- EOLO-Ca) em juvenis de tambaqui (Colossoma macropomum). As concentrações testadas dos EOs de L. alba e L. origanoides foram: 25, 50, 100 ou 200 μL/L. O tempo para atingir os estágios de sedação, anestesia profunda, recuperação e frequência ventilatória foram monitorados em cada concentração de EO. Os resultados indicam que todas as concentrações testadas dos OE induziram a anestesia do tambaqui e que a frequência ventilatória foi alterada pelos EOs. L. origanoides mostrou o tempo mais rápido para induzir sedação e anestesia, mas os tempos de recuperação foram longos. EO L. alba (citral 100 μL/L e linalol 200 μL/L) pode ser utilizado como anestésico em juvenis de tambaqui, pela eficiência em anestesiar e recuperar.

The essential oil of Lippia alba and its components affect Drosophila behavior and synaptic physiology.

Lippia alba is a flowering shrub in the verbena family and its essential oil (EO) is known for its sedative, antidepressant and analgesic properties. In the Amazon region of Brazil, it is used in aquaculture to anesthetize fish during transport. Many of the specialized metabolites found in EOs presumably evolved to protect plants from herbivores, especially insects. We used Drosophila to test the behavioral and physiological actions of this EO and its components. We found that a 150 min exposure to the EO vapors resulted in immobilization of adult flies. Gas chromatography-mass spectrometry identified the major components of the EO as the monoterpenes citral (59%), carvone (7%) and limonene (7%). Fly immobilization by the EO was due to citral and carvone, with citral producing more rapid effects than carvone. We tested whether the EO affected synaptic physiology by applying it to the larval neuromuscular junction. The EO delivered at 0.012% (v/v) produced over a 50% reduction in excitatory postsynaptic potential (EPSP) amplitude within 3-4 min. When the EO components were applied at 0.4 mmol l-1, citral and carvone produced a significant reduction in EPSP amplitude, with citral producing the largest effect. Measurement of miniature EPSP amplitudes demonstrated that citral produced over a 50% reduction in transmitter release. Calcium imaging experiments showed that citral produced about 30% reduction in presynaptic Ca2+ influx, which likely resulted in the decrease in transmitter release. Thus, the EO blocks synaptic transmission, largely due to citral, and this likely contributes to its behavioral effects.

Citral and linalool chemotypes of Lippia alba essential oil as anesthetics for fish: a detailed physiological analysis of side effects during anesthetic recovery in silver catfish (Rhamdia quelen).

The viability using Lippia alba essential oil as an anesthetic for fish was studied, particularly with respect to physiological effects during recovery. Anesthesia of silver catfish (Rhamdia quelen) using 100 and 300 µL L-1 of two different chemotypes of L. alba essential oil (citral EO-C and linalool EO-L) prevented the increase of plasma cortisol levels caused by handling, but did not avoid alterations in energetic metabolism. Silver catfish did not have increased the levels of thiobarbituric acid reactive species in the kidney and liver during recovery after anesthesia with either EO, avoiding lipid damage. On the other hand, fish anesthetized with EO-C showed higher protein carbonylation levels, superoxide dismutase, catalase, and glutathione S-transferase activities and non-protein thiol group levels in both tissues compared to controls. Our results suggest that both oils show antioxidant capacity, but anesthesia with EO-L does not cause damage to lipids or proteins, only temporary changes, typical of physiological adjustments during recovery from anesthesia. Therefore, EO-L is an effective anesthetic for silver catfish with fewer side effects than EO-C.

Anesthesia and sedation of map treefrog (Hypsiboas geographicus) tadpoles with essential oils / Anestesia e sedação de girinos da perereca geográfica Hypsiboas geographicus com óleos essenciais

ABSTRACT: The goal of this study was to investigate the sedative and anesthetic properties of essential oils (EOs) in map treefrog tadpoles (Hypsiboas geographicus) and to determine the sedation and deep anesthesia induction times as well as the recovery time. The tadpoles were exposed to one of the EOs from three plant species: Aniba rosaeodora (EOAR - 25, 50, 100 or 200µL L-1), Lippia origanoides (EOLO - 13, 25, 50, 100 or 200µL L-1), and Lippia alba (either chemotype citral [EOL-C - 25, 50, 100 or 200µL L-1] or linalool [EOL-L - 50, 75, 100 or 200µL L-1]) (n = 8 per replicate). The tadpoles exposed to 25 and 50µL L-1 EOL-C and EOL-L, respectively, were not anesthetized within 30min (the maximum time of observation), and those exposed to 200µL L-1 EOLO did not recover within 30min. Sedation, deep anesthesia and recovery times showed a concentration-dependent relationship for all EOs tested, with the exception of the recovery with EOLO. The results allowed concluding that all investigated EOs can be used to anesthetize tadpoles of H. geographicus, but the use of EOLO must not exceed 100µL L-1.

RESUMO: O objetivo deste estudo foi investigar as propriedades sedativas e anestésicas de óleos essenciais (OEs) em girinos da perereca Hypsiboas geographicus e determinar os tempos de indução à sedação e anestesia profunda, bem como o de recuperação. Os girinos foram expostos a um dos OEs de três espécies de plantas: Aniba rosaeodora (OEAR - 25, 50, 100 ou 200µL L-1), Lippia origanoides (OELO - 13, 25, 50, 100 ou 200µL L-1 ) ou Lippia alba quimiotipos citral (OEL-C - 25, 50, 100 ou 200µL L-1) ou linalol (OEL-L - 50, 75, 100 ou 200µL L-1) (n = 8 cada repetição). Girinos expostos a 25 e 50µL L-1 OEL-C e OEL-L, respectivamente, não foram anestesiados dentro de 30min (tempo máximo de observação) e aqueles expostos a 200µL L-1 OELO não recuperaram dentro de 30min. Os tempos de sedação, anestesia profunda e recuperação apesentaram uma relação concentração-resposta para todos os OEs testadas, exceto a recuperação com OELO. Os resultados permitem concluir que todos os OEs investigados podem ser usados para anestesiar os girinos de H. geographicus, mas o uso de OELO não deve ser superior a 100µL L-1.

Trypanocidal action of Lippia alba and Lippia origanoides essential oils against Trypanosoma evansi in vitro and in vivo used mice as experimental model.

Parasitic diseases have an enormous health and economic impact and are a particular problem in tropical regions of the world. Disease caused by protozoa, such as trypanosomiasis, are the cause of most parasite related morbidity and mortality. Thus, the aim of this study was to verify the trypanocidal effectiveness of Lippia alba and Lippia origanoides against Trypanosoma evansi in vitro and in vivo. L. alba and L. origanoides were used in vitro on trypomastigotes at different concentrations (0.5, 1.0 and 2.0 %) and exposure times (0, 1, 3, 6 and 9 h). The three concentrations tested showed trypanocidal activity in vitro, completely eliminating the parasites in small concentration after 6 h of assay. In vivo tests were performed using mice as the experimental model. T. evansi infected mice were treated with L. alba and L. origanoides with dose of 1.5 mL kg-1 during 5 days. These protocols did not provide curative efficacy, however the mice treated with L. origanoides showed a significant increase in the longevity when compared to control group. Active compounds present in essential oils, such as L. origanoides, may potentiate the treatment of trypanosomosis when associated with other trypanocidal drugs.

Physiological responses of Rhamdia quelen (Siluriformes: Heptapteridae) to anesthesia with essential oils from two different chemotypes of Lippia alba

The aim of this study was to evaluate if Lippia alba has different chemotypes according to the chemical composition of the essential oil (EO) considering collection site, and if the EO may have different effects on blood and plasma parameters in silver catfish, Rhamdia quelen, during and immediately after anesthesia. The citral (EO-C) and linalool (EO-L) chemotypes were identified, and both presented similar anesthetic effects for silver catfish. Fish were exposed to two concentrations of each EO, which induced slow and fast anesthesia (100 and 300 µL L-1, respectively). Blood ions did not change at any time of anesthesia induction and recovery and, therefore, the electrolyte balance was not altered. Blood gases oscillated through all exposure and recovery times, but there was an increase in pO2 after 10 min recovery in fish anesthetized with EO-C. Glucose increased in fish exposed to both EOs when compared with the control group. Overall, exposure to both EOs (except 100 µL L-1 EO-L at most times) reduced plasma cortisol levels compared to the control and/or ethanol groups. However, as plasma creatinine levels in fish anesthetized with EO-C were higher than control fish, the use of EO-L is preferable.(AU)

O objetivo deste estudo foi avaliar se Lippia alba apresenta diferentes quimiotipos de acordo com a composição química do óleo essencial (OE), considerando local de coleta e se o OE causa diferentes efeitos nos parâmetros sanguíneos e plasmáticos em jundiá, Rhamdia quelen, durante e imediatamente após a anestesia. Os quimiotipos citral (OE-C) e linalol (OE-L) foram identificados e ambos apresentaram efeito anestésico semelhante para jundiá. Os peixes foram expostos a duas concentrações de cada OE, que induziram anestesia lenta e rápida (100 e 300 mL L-1, respectivamente). Íons sanguíneos não se alteraram em nenhum tempo e consequentemente, o equilíbrio eletrolítico não foi alterado. Os gases sanguíneos oscilaram durante todo tempo de exposição e recuperação, mas houve aumento na pO2 após 10 min de recuperação em peixes anestesiados com OE-C. Níveis sanguíneos de glicose aumentaram nos peixes expostos a ambos OEs quando comparados com o grupo controle. De um modo geral, a exposição a ambos OEs (exceto 100 µL L-1 OE-L na maioria dos tempos) reduziu o cortisol plasmático comparado aos grupos controle e etanol. No entanto, como os níveis de creatinina plasmática em peixes anestesiados com OE-C foram maiores que nos peixes controle, é preferível o uso do OE-L.(AU)

Are similar the parasite communities structure of Trachelyopterus coriaceus and Trachelyopterus galeatus (Siluriformes: Auchenipteridae) in the Amazon basin?

The aim of this study was to compare the parasite communities in two sympatric host populations, Trachelyopterus coriaceus andTrachelyopterus galeatus, which were caught in tributary of the Amazon River in Brazil. All the specimens of T. galeatusand T. coriaceus were infected by one or more parasites, such as Ichthyophthirius multifiliis, Tripartiella tetramerii, Trichodina nobilis, Cosmetocleithrum striatuli, Contracaecumsp., Cystidicoloides sp., Dadaytremoides parauchenipteri and Gorytocephalus spectabilis. Seven species were common to both host fish, and there were 1-5 parasite species per host. In both hosts, trichodinids were dominant. Aggregate dispersion of ectoparasites and endoparasites was observed, with greater aggregation among endoparasites. Only the ectoparasites species showed differences in intensity and/or abundance. However, the parasite communities of the two hosts were taxonomically similar (99%) and characterized by high prevalence and high abundance of ectoparasites, but with low diversity, prevalence and abundance of endoparasites. Trachelyopterus galeatus, the host with the larger body size, presented greater variation of Brillouin diversity and evenness, while T. coriaceus had higher Berger-Parker dominance values and total numbers of parasites. This first study on these parasites ofT. galeatus and T. coriaceus showed that the life mode, size of the hosts and the availability of infective forms of the parasites were the main factors that influenced the parasite communities structure.

Are similar the parasite communities structure ofTrachelyopterus coriaceus and Trachelyopterus galeatus (Siluriformes: Auchenipteridae) in the Amazon basin? / São similares a estrutura das comunidades de parasitos deTrachelyopterus coriaceus e Trachelyopterus galeatus (Siluriformes: Auchenipteridae) na bacia amazônica?

Abstract The aim of this study was to compare the parasite communities in two sympatric host populations, Trachelyopterus coriaceus andTrachelyopterus galeatus, which were caught in tributary of the Amazon River in Brazil. All the specimens of T. galeatusand T. coriaceus were infected by one or more parasites, such as Ichthyophthirius multifiliis, Tripartiella tetramerii, Trichodina nobilis,Cosmetocleithrum striatuli, Contracaecumsp., Cystidicoloides sp., Dadaytremoides parauchenipteri and Gorytocephalus spectabilis. Seven species were common to both host fish, and there were 1-5 parasite species per host. In both hosts, trichodinids were dominant. Aggregate dispersion of ectoparasites and endoparasites was observed, with greater aggregation among endoparasites. Only the ectoparasites species showed differences in intensity and/or abundance. However, the parasite communities of the two hosts were taxonomically similar (99%) and characterized by high prevalence and high abundance of ectoparasites, but with low diversity, prevalence and abundance of endoparasites. Trachelyopterus galeatus, the host with the larger body size, presented greater variation of Brillouin diversity and evenness, while T. coriaceus had higher Berger-Parker dominance values and total numbers of parasites. This first study on these parasites ofT. galeatus and T. coriaceus showed that the life mode, size of the hosts and the availability of infective forms of the parasites were the main factors that influenced the parasite communities structure.

Resumo O objetivo deste estudo foi comparar as comunidades, prevalência, intensidade, abundância e interações parasito-hospedeiro em duas populações de hospedeiros simpatricos, Trachelyopterus coriaceus eTrachelyopterus galeatus de um tributário do sistema do Rio Amazonas, no Brasil. Todos os espécimes de T. galeatus eT. coriaceus estavam infectados por um ou mais parasitos, tais como Ichthyophthirius multifiliis, Tripartiella tetramerii, Trichodina nobilis,Cosmetocleithrum striatuli, Contracaecumsp., Cystidicoloides sp., Dadaytremoides parauchenipteri e Gorytocephalus spectabilis. Sete espécies de parasitos foram comuns para ambos os peixes hospedeiros e houve de 1-5 espécies de parasitos por hospedeiro. Em ambos os hospedeiros houve dominância de espécies de tricodinídeos. A dispersão dos ectoparasitos e endoparasitos foi agregada, mas a maior agregação foi de endoparasitos. Somente espécies de ectoparasitos mostraram diferenças na intensidade e/ou abundância. Entretanto, as comunidades parasitárias dos dois hospedeiros foram taxonomicamente semelhante (99%) e caracterizada por alta prevalência e abundância de ectoparasitos, mas com baixa diversidade, prevalência e abundância de endoparasitos. Trachelyopterus galeatus, com maior tamanho corporal, apresentou maior variação da diversidade Brillouin e uniformidade, enquanto T. coriaceus teve maiores valores de dominância de Berger-Parker e número total de parasitos. Este primeiro estudo sobre estes parasitos de T.galeatus e T. coriaceus mostrou que o modo de vida, tamanho dos hospedeiros e disponibilidade de formas infectantes de parasitos foram os principais fatores que influenciaram a estrutura das comunidades dos parasitos encontrados.

A social and ecological assessment of tropical land uses at multiple scales: the Sustainable Amazon Network.

Science has a critical role to play in guiding more sustainable development trajectories. Here, we present the Sustainable Amazon Network (Rede Amazônia Sustentável, RAS): a multidisciplinary research initiative involving more than 30 partner organizations working to assess both social and ecological dimensions of land-use sustainability in eastern Brazilian Amazonia. The research approach adopted by RAS offers three advantages for addressing land-use sustainability problems: (i) the collection of synchronized and co-located ecological and socioeconomic data across broad gradients of past and present human use; (ii) a nested sampling design to aid comparison of ecological and socioeconomic conditions associated with different land uses across local, landscape and regional scales; and (iii) a strong engagement with a wide variety of actors and non-research institutions. Here, we elaborate on these key features, and identify the ways in which RAS can help in highlighting those problems in most urgent need of attention, and in guiding improvements in land-use sustainability in Amazonia and elsewhere in the tropics. We also discuss some of the practical lessons, limitations and realities faced during the development of the RAS initiative so far.

Lethal concentration of clomazone, metsulfuron-metil, and quinclorac for silver catfish, Rhamdia quelen, fingerlings

O objetivo deste trabalho foi determinar a concentração letal (CL50 96h) de clomazone, metsulfuron-metil e quinclorac, herbicidas utilizados no cultivo de arroz, para alevinos de jundiá, Rhamdia quelen. Alevinos foram expostos a diferentes concentrações destes herbicidas. A CL50 foi 7,32µL L-1 para clomazone e 395mg L-1 para quinclorac. A CL50 para metsulfuron-metil não foi encontrada porque todos os alevinos sobreviveram mesmo à concentração de 1200mg L-1. Provavelmente apenas o clomazone pode provocar alguma mortalidade em jundiás mantidos em rizipiscicultura.