Cavefish cope with environmental hypoxia by developing more erythrocytes and overexpression of hypoxia-inducible genes.

Dark caves lacking primary productivity can expose subterranean animals to hypoxia. We used the surface-dwelling (surface fish) and cave-dwelling (cavefish) morphs of Astyanax mexicanus as a model for understanding the mechanisms of hypoxia tolerance in the cave environment. Primitive hematopoiesis, which is restricted to the posterior lateral mesoderm in other teleosts, also occurs in the anterior lateral mesoderm in Astyanax, potentially pre-adapting surface fish for hypoxic cave colonization. Cavefish have enlarged both hematopoietic domains and develop more erythrocytes than surface fish, which are required for normal development in both morphs. Laboratory-induced hypoxia suppresses growth in surface fish but not in cavefish. Both morphs respond to hypoxia by overexpressing hypoxia-inducible factor 1 (hif1) pathway genes, and some hif1 genes are constitutively upregulated in normoxic cavefish to similar levels as in hypoxic surface fish. We conclude that cavefish cope with hypoxia by increasing erythrocyte development and constitutive hif1 gene overexpression.

Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report / Myxobolus spp. (Cnidaria: Myxobolidae) no sangue circulante de peixes dos estados de Goiás e Mato Grosso, Brasil: relato de caso

Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

A mixosporidiose é considerada uma doença infecciosa causada por mixozoários pertentences ao Filo Cnidaria, Classe Myxosporea e Ordem Bivalvulida, considerados parasitos comuns de peixes de água doce e salgada, parasitando vários órgãos, principalmente as brânquias. No presente estudo, 49 espécimes de peixes pertencentes a oito gêneros: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus and Serrasalmus foram coletados e extensões sanguíneas foram feitas com a finalidade de encontrar hemoparasitos. Entretanto, mixoesporos foram observados em cinco (10.20%) espécimes de peixes, pertencentes aos gêneros Tetragonopterus, Myleus e Pygocentrus. Dois morfotipos de Myxobolus spp. foram identificados parasitando esses peixes. Normalmente, as investigações sobre mixozoários em peixes é realizada com a procura de plasmódios ou cistos nos órgãos dos peixes e observação da cavidade de órgãos. Porém, esse trabalho ressalta a importância de se examinar também o sangue desses animais, já que esses parasitos podem ocasionar patogenias severas em peixes. Sendo assim, a análise sanguínea pode proporcionar controle sanatório preventivo para peixes comerciais e evitar perdas econômicas.

Sex disparities determined by biometric and hematological measurements in Astyanax bimaculatus / [Disparidades sexuais determinadas por medições biométricas e hematológicas em Astyanax bimaculatus]

This study aimed to compare the body indexes and hematological characteristics between Astyanax bimaculatus males and females. Four hundred fish were randomly distributed into four polyethylene tanks (100 fish/unit) in a recirculation system and fed four times a day (3% of biomass). After 90 days, ten fish (five ♀ and five ♂) were removed to perform blood tests and to measure weight, height, total length, height/length ratio, condition factor and index determination: vicerosomatic (VSI), hepatosomatic (HSI), and gonadosomatic (GSI). The results showed a higher average weight (g) in females (12.32±0.71) compared to males (6.98±0.75), the same happened to height (cm) = (3.01±0.07) and (2.40±0.05), total length (cm) = (3.01±0.07) and (2.40±0.05), VSI (%) = (11.43±0.81) and (3.55±1.05), HSI (%) = (0.72±0.08) and (0.30±0.04), respectively. Mean corpuscular hemoglobin (pg) was higher in females (3.72±1.20) than in males (2.99±1.51). Regarding the number of thrombocytes (103.µL-1), there was an increase in males (25.71±3.91) compared to females (17.40±6.40).(AU)

O objetivo deste trabalho foi comparar os índices corporais e as características hematológicas entre machos e fêmeas de Astyanax bimaculatus. Quatrocentos peixes foram distribuídos aleatoriamente em quatro caixas de polietileno (100 peixes/unidade), em sistema de recirculação, e alimentados quatro vezes ao dia (3% da biomassa). Após 90 dias, 10 peixes (cinco ♀ e cinco ♂) foram retirados para realização das análises sanguíneas e para mensuração do peso, da altura, do comprimento total, da relação altura/comprimento, do fator de condição e da determinação dos índices: viscerossomático (IVS), hepatossomático (IHS) e gonadossomático (IGS). Os resultados mostraram um maior peso médio (g) nas fêmeas (12,32±0,71) em relação aos machos (6,98±0,75); o mesmo aconteceu para altura (cm) = (3,01± 0,07) e (2,40± 0,05), comprimento total (cm) = (3,01±0,07) e (2,40±0,05), IVS (%) = (11,43±0,81) e (3,55±1,05), IHS (%) = (0,72±0,08) e (0,30±0,04), respectivamente. Hemoglobina corpuscular média (pg) foi maior nas fêmeas (3,72±1,20) que nos machos (2,99±1,51). Em relação ao número de trombócitos (103/µL), houve um aumento nos machos (25,71± 3,91) em relação às fêmeas (17,40±6,40).(AU)

Probiotic supplementation causes hematological changes and improves non-specific immunity in Brycon amazonicus

A commercialprobioticcontaining Bacillussubtilis(109 CFU g-1) was evaluated incaged matrinxã,Bryconamazonicus,by measuring hematological parametersand macrophage activity after 42 and 84 days after feeding. The product wasadded tocommercial feed using 2%soybean oil as a protectant. A randomized three-treatmentexperiment was performed using fourreplicates per treatment. The groups included: (a) control without probiotic, (b) 5 g kg-1probiotic, and (c) 10 g kg-1probiotic. Forhematological analysis,eightfishper treatmentwere used to determinetotal cell count (RBC); thrombocytes, differential, and total leukocyte count (TLC); hematocrit (Htc); hemoglobin tax; mean corpuscular volume (MCV); and mean corpuscular hemoglobin concentration (MCHC). Furthermore, plasma cortisol and glucose levels were measured in blood samples. Macrophage phagocytic activity was evaluated by injecting Saccharomyces cerevisiae(11,000 cells in a 3 mLvolume) into the coelomic cavity incubating for 8hours.Addition of probiotics to the diet of caged matrinxã altered the Htc, RBC, MCV, MCHC, TLC, lymphocyte, and eosinophil values. We observed increased cortisol and glucose levels and phagocytic activity, but no increase in the phagocytic index. We thus conclude that supplementing caged Brycon amazonicuswith probiotics improves their non-specific immunity and alters blood profiles.

Thermal manipulation and GnRHa therapy applied to the reproduction of lambari-do-rabo-amarelo, Astyanax altiparanae females (Characiformes: Characidae) during the non-breeding season.

Lambari-do-rabo-amarelo Astyanax altiparanae in the wild reproduce during spring and summer, but females undergo vitellogenesis throughout the year, including the non-spawning winter period when water temperatures are low. The present study investigated the physiological role of temperature modulation on the hypothalamus-pituitary-gonads axis of lambari during winter, as well as the effects of gonadotropin releasing hormone agonist (GnRHa) therapy. Captive females were exposed to two different temperatures (20 °C and 27 °C) and were injected weekly with GnRHa for 21 days during winter (Control, CTR; Low dose; LD and high dose of GnRHa, HD). At the end of the 21-days period gonadosomatic index (GSI), oocyte stage of development and theoretical fecundity were evaluated, together with plasma levels of 17ß-estradiol (E2). Gene expression of the two pituitary gonadotropins follicle-stimulating hormone (fshß) and luteinizing hormone (lhß), as well as hepatic vitellogenin-A (vtgA) expression were also analyzed. At the end of the experimental period, females from the six different experimental conditions were induced to spawn using human chorionic gonadotropin (hCG). Spawning performance parameters and plasma levels of the maturation inducing steroid (MIS) were analyzed. Gene expression of fshß did not change with temperature manipulation, but females exposed to 27 °C and supplemented with a HD of GnRHa exhibited an increased fshß gene expression, associated with higher E2 levels. The higher water temperature alone was able to increase E2 levels. At both water temperatures GnRHa injections induced a decrease in E2 levels. GnRHa injected females had a lower vtgA gene expression levels at 20 °C. Even with differences in the gene expression of gonadotropins among the various temperature/GnRHa treatments, GSI and oocyte diameter did not change, but GnRHa enhanced the number of vitellogenic oocytes at 20 °C. The reproductive performance of lambari induced to spawn with hCG was better after the combined treatment with GnRHa and summer temperature.

Evaluación genotóxica del agua del Río Grande (Antioquia, Colombia) mediante frecuencia de eritrocitos micronucleados de Brycon henni (Characiformes: Characidae) / Genotoxic evaluation of Río Grande (Antioquia, Colombia) water using micronucleus frequency in erythrocytes of Brycon henni (Characiformes: Characidae)

ResumenLa producción agrícola de monocultivo permanente de banano y plátano en la parte media del Río Grande (Turbo - Antioquia) requiere de la aplicación de diferentes plaguicidas. Prácticas inapropiadas en la producción y cultivos de plátano realizadas en esta región a menudo conllevan a contaminación con agroquímicos que llegan a este cuerpo de agua por procesos de lixiviación y escorrentía. Los peces son los vertebrados más utilizados como especies bioindicadoras de la calidad del agua porque son muy sensibles a la presencia de contaminantes en el ambiente. El objetivo de esta investigación fue comparar la frecuencia de eritrocitos micronucleados (EMN) en sangre periférica de ejemplares de peces sabaleta (Brycon henni) procedentes de dos estaciones del Río Grande que difieren en su grado de contaminación por agroquímicos. Se evaluó la frecuencia de EMN en sangre periférica de peces B. henni procedentes de cada estación durante dos épocas de lluvia de 2010 y dos época secas de 2011. Las muestras de sangre fueron obtenidas de la vena caudal, fijadas durante 24 horas y luego teñidas con Giemsa. La frecuencia de EMN fue significativamente mayor en el área impactada por agroquímicos. La frecuencia media de EMN fue mayor en la estación con impacto directo de agroquímicos (0.15±0.18) que en la estación poco alterada (0.06±0.08). Además, la frecuencia de EMN en B. henni fue significativamente mayor en la época seca. Los resultados de este estudio indican que el análisis de las EMN en B. henni puede ser recomendada como un método adecuado para la detección in situ de agentes genotóxicos en el ambiente.

Abstract:The permanent monoculture of bananas and plantains farming in the middle of Río Grande (Turbo - Antioquia) requires the application of a variety of pesticides. Inappropriate banana production practices in this region, have often led to waterbody pollution by agrochemicals from leachate and runoff processes. Currently, fish are the most common vertebrates used as bioindicators of water quality, because they are very sensitive to the presence of contaminants. Our main goal with this study was to compare the frequency of micronucleated erythrocytes (MNE) in peripheral blood specimens of fish Brycon henni, from two locations (polluted and unpolluted) in the Rio Grande. We evaluated the frequency of MNE in peripheral blood samples of fish B. henni from each location during two rainy seasons in 2010 and two dry seasons in 2011. Blood samples were collected, fixed for 24 h, and then were stained with Giemsa. Among results, we found that the median frequency of MNE was higher in the polluted site by agrochemical discharges (0.15±0.18), than in the unimpacted site (0.06±0.08). Furthermore, the frequency of MNE in B. henni during the dry season was highly significant for both locations. The results of this study indicated that the analysis of MNE in B. henni could be recommended as a suitable method for in situ detection of environmental genotoxins. Rev. Biol. Trop. 65 (1): 405-414. Epub 2017 March 01.

Hematological responses of tambaqui Colossoma macropomum (Serrassalmidae) fed with diets supplemented with essential oil from Mentha piperita (Lamiaceae) and challenged with Aeromonas hydrophila / Respostas hematológicas de tambaqui Colossoma macropomum (Serrassalmidae) alimentado com dietas suplementadas com óleo essencial de Mentha piperita (Lamiaceae) e desafiados com Aeromonas hydrophila

In fish farmings, diseases can be reduced by using immunostimulants. The aim of this study was to evaluate the immunostimulant potential of Mentha piperita in tambaqui fed with 0, 0.5, 1.0 and 1.5% of oil per kg of commercial fish feed. The fish were inoculated with Aeromonas hydrophila to challenge them. Hematological and biochemical parameters were determined after 30 days of feeding and seven days after the challenge. There was no mortality and M. piperita oil did not influence fish production parameters. However, blood hemoglobin concentration (Hb) increased in the fish fed with 0.5 and 1.5% of oil per kg of diet; albumin increased in those fed with 1.0%; cholesterol increased in all groups with oil; and triglycerides increased in those fed with 0.5%. After the bacterial challenge, the fish showed decreases in Hb when fed with diet enriched with 1.5% oil per kg of diet, in mean corpuscular volume with 1.0% and in mean corpuscular hemoglobin concentration with 0 and 1.5%. Protein levels increased in groups with 0 and 1.5% of oil and albumin when fed with 0 and 1.0%; cholesterol levels increased in the control group; and high levels of triglycerides were observed in the groups with 0, 0.5 and 1.5%. Thus, M. piperita essential oil promoted hematological alterations in tambaqui and can be recommended in diets containing up to 1.0% per kg, because of the minimal physiological modifications caused. However, additional studies are necessary to obtain more information regarding to the physiological effects of this immunostimulant.

Em pisciculturas, doenças podem ser minimizadas com o uso de imunoestimulantes. O objetivo deste estudo foi avaliar o potencial imunoestimulante do óleo essencial de Mentha piperita na dieta de tambaqui com dietas contendo 0; 0,5; 1,0; 1,5% de inclusão do óleo por kg de ração comercial para peixes. Os tambaquis foram desafiados com a bactéria Aeromonas hydrophila. Hematologia e bioquímica dos tambaquis foram determinados após 30 dias de alimentação e sete dias do desafio. Não houve mortalidade dos peixes durante o experimento, sendo que o óleo não influenciou os parâmetros produtivos dos peixes. Entretanto, a concentração de hemoglobina (Hb) aumentou em peixes alimentados com 0,5 e 1,5% de óleo por kg de dieta; a albumina aumentou em 1,0%; o colesterol aumentou em todos os grupos com óleo; e triglicérides em 0,5% por kg de dieta. Após desafio bacteriano ocorreram diminuições da Hb nos peixes alimentados com 1,5% de óleo; do volume corpuscular médio com 1,0%; e da concentração de hemoglobina corpuscular média com 0 e 1,5%. Níveis aumentados de proteína nos grupos 0 e 1,5%; da albumina em 0 e 1,0%; do colesterol no controle; e dos triglicérides em 0, 0,5 e 1,5% de óleo na dieta. Assim o óleo essencial de M. piperita na dieta promoveu alterações hematológicas e pode ser recomendado seu uso em inclusões de até 1,0% por kg da dieta de tambaqui, devido às mínimas alterações fisiológicas causadas. Entretanto, estudos adicionais são necessários, de modo a obter informações sobre os efeitos fisiológicos deste imunoestimulante.

Haematological and histopathological analysis in South American fish Piaractus mesopotamicus parasitized by monogenean (Dactylogyridae) / Análises hematológica e histopatológica no peixe sul-americano Piaractus mesopotamicus parasitado por Monogenea (Dactylogyridae)

Monogeneans are the parasites mostly found on the body surface and gills of fish and can cause large losses in farmed fish. Some studies demonstrate elevated parasitic levels causing hematological alterations. But few of them relate the effects of parasitism on the hematology and histopathology of native freshwater farmed fish. This study evaluated the host-parasite relationship in pacu (Piaractus mesopotamicus) parasitized by the monogenean Anacanthorus penilabiatus. Hematological and parasitological assessments were obtained in 60 fish captured in a fish farm located in Dourados, State of Mato Grosso do Sul, Central Brazil. Fish were analyzed in different categories of parasite number: class I (n=13; 0-200 parasites), class II (n=17; 201-1200 parasites); class III (n=7; 1201-2200 parasites); and class IV (n=23; more than 2200 parasites per host). The highest levels of parasitism caused significant decrease (p<0.05) in the hematocrit, red blood cells (RBC), mean hemoglobin concentration (MCHC) and basophils number. Thrombocytes, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), monocytes, eosinophils, neutrophils and LG-PAS did not present significant difference among the parasitic levels. In contrast, increased number of total leukocytes and lymphocytes were found in highly-parasitized fish. A positive linear correlation (p<0.01) was found between the amount of parasites and fish weight. Histopathology revealed severe hyperplasia, sub-epithelial edema, fusion of the secondary lamellae, focal and multifocal necrosis in highly parasitized fish.

Parasitos Monogenea são principalmente encontrados na superficie corporal e brânquias dos peixes, e podem acarretar grandes perdas em pisciculturas. Alguns estudos demonstram que elevados níveis de infestação parasitária podem alterar os parâmetros sanguíneos. Porém, poucos estudos se direcionam a esclarecer os efeitos do parasitismo sobre as características hematológicas em peixes nativos. Este estudo avaliou a relação parasito-hospedeiro em pacu (Piaractus mesopotamicus) parasitado pelo monogenético Anacanthorus penilabiatus. Avaliações hematológicas e parasitológicas foram obtidas de 60 peixes capturados de uma piscicultura localizada em Dourados, Estado de Mato Grosso do Sul (MS), Brasil Central. Os peixes foram divididos em diferentes categorias de número de parasitos: classe I (n=13; 0-200 parasitos), classe II (n=17; 201-1200 parasitos); classe III (n=7; 1201-2200 parasitos); e classe IV (n=23; mais que 2200 parasitos por hospedeiro). Os níveis mais elevados de parasitismo causaram diminuição significativa (p<0.05) no hematócrito, eritrócitos (RBC), concentração de hemoglobina corpuscular média (CHCM) e número de basófilos. Trombócitos, volume corpuscular médio (VCM), concentração de hemoglobina corpuscular media (CHCM), monócitos, eosinófilos, neutrófilos e LG-AS não apresentaram diferença significativa entre os níveis de parasitismo. Em contraste, o aumento do número de leucócitos totais e linfócitos foram encontrados em peixes altamente parasitados. Houve correlação linear positiva entre a quantidade de parasitos e o peso dos peixes. O exame histopatológico revelou severa hiperplasia, edema sub-epitelial, fusão das lamelas secundárias, necroses focal e multifocal em peixes altamente parasitados.