Gross, histologic, and ultrastructural features of iridophoromas in Siamese fighting fish (Betta splendens).

Pigment-containing and light-reflecting cell neoplasms, generically termed chromatophoromas, affect fish, reptiles, and amphibians. Chromatophoromas of light-reflecting cells are named iridophoromas. In this study, we aimed to describe the gross, histologic, and ultrastructural findings of 71 cases of iridophoromas in farmed Siamese fighting fish (Betta splendens). Macroscopically, iridophoromas appeared as whitish, gray, or black friable masses or plaques in the fin, trunk/tail, or head of the fish. Forty-five tumors (63%) were malignant and invaded the adjacent skeletal muscle and/or metastasized to other organs, whereas 26 (37%) tumors were restricted only to the skin, but due to the cytologic similarity to the malignant counterpart, we were not able to classify them as malignant or benign. Sixty-five (91%) tumors were classified as iridophoromas, whereas 6 (8%) were diagnosed as mixed chromatophoromas. Despite immunolabeling for PNL-2, melan A, or S-100 failing to demonstrate antigen expression, ultrastructural analysis identified light-reflecting neoplastic cells, unequivocally confirming iridophoromas as the predominant tumor. The high incidence of iridophoromas in Siamese fighting fish from the same breeding facility, coupled with a higher occurrence in royal blue and fancy copper color patterns and in young males, suggests a potential genetic/hereditary factor in the tumorigenesis of these neoplasms.

Effect of temperature on the embryonic and larvae development of discus fish Symphysodon aequifasciatus and time of first feeding.

This study aimed to analyze the influence of different temperatures on the embryonic and larval development of discus fish Symphysodon aequifasciatus and determine the time required for the beginning of exogenous feeding. Eggs and larvae were obtained from natural spawns and distributed in five treatments: 24.0, 26.0, 28.0, 30.0, and 32.0 °C. To assess the developmental stages and embryonic structures, samples were taken at regular intervals and checked under an optical microscope. At the end of the experimental period, statistical analysis was performed, followed by Tukey's test. As a result, it was possible to observe the significant effects of temperature on the variables. It was noted that the temperature accelerated the embryonic and larval development of the discus and also contributed to a reduction in the time between the incubation period and the feeding transition. It was also noted that the incubation of eggs and larvae at a temperature of 24.0 °C can cause damage to embryos, such as malformation of the body as well as anomalies in the circulatory system.

Antimicrobial and anthelmintic effects of copper nanoparticles against Koi carp parasites and their toxicity.

This study investigated the in vitro antimicrobial and anthelmintic effect of copper nanoparticles (CuNPs) against the bacterium Aeromonas hydrophila, the monogeneans Dactylogyrus minutus, Dactylogyrus extensus, Gyrodactylus cyprini, and the cestode Schyzocotyle acheilognathi, as well as their toxicity to Cyprinus carpio Koi. In the antimicrobial in vitro test, the inhibition zone method and minimum inhibitory concentration (MIC) were performed. In order to determine the time and efficacy of monogenean parasite mortality, the parasites were exposed to CuNP concentrations of 20, 50, 100, 150, 200, and 300 mg L-1, and a control group with tank water and one with copper sulphate pentahydrate (CuSO4.5H2O) at a concentration of 0.3 mg L-1, performed in triplicate. The parasites were observed every 10 min for 300 min, and mortality was recorded. For the cestodes, parasites were immersed in CuNP concentrations of 50, 100, 150, and 300 mg L-1. At the end of the in vitro tests, the anthelmintic efficacy of each treatment was calculated. To assess the tolerance and toxicity in fish, they were exposed to CuNP concentrations of 0.6, 1.25, 2.5, 5, 10, 20, and 50 mg L-1 for 12 h. The MIC demonstrated that CuNPs effectively inhibited the growth of A. hydrophila up to a dilution of 12,500 mg L-1 and showed an inhibition zone of 14.0 ± 1.6 mm for CuNPs. The results of anthelmintic activity showed a dose-dependent effect of concentration for both groups of parasites, with the most effective concentration being 300 mg L-1 in 120 min. In the toxicity test, the carps showed tolerance to lower concentrations. The study indicated that CuNPs were effective against the studied pathogens. However, it proved to be toxic to fish at high concentrations. The use of low concentrations is recommended still requires further investigation.

Effect of monocultures and polycultures of Typha latifolia and Heliconia psittacorum on the treatment of river waters contaminated with landfill leachate/domestic wastewater in partially saturated vertical constructed wetlands.

Partially Saturated Vertical Constructed Wetlands (PSV-CWs) are novel wastewater treatment systems that work through aerobic and anaerobic conditions that favor the removal of pollutants found in high concentrations, such as rivers contaminated with domestic wastewater and landfill leachate. The objective of the study was to evaluate the efficiency of PSV-CWs using monocultures and polycultures of Typha latifolia and Heliconia psittacorum to treat river waters contaminated with leachates from open dumps and domestic wastewater. Six experimental units of PSV-CWs were used; two were planted with Typha latifolia monoculture, two with Heliconia psittacorum monoculture and two with polycultures of both plants. The results indicated better organic matter and nitrogen removal efficiencies (p < 0.05) in systems with polycultures (TSS:95%, BOD5:83%, COD:89%, TN:82% and NH4+:99%). In general, the whole system showed high average removal efficiencies (TSS:93%, BOD5:79%, COD:85%, TN:79%, NH4+:98% and TP:85%). Regarding vegetation, both species developed better in units with monocultures, being Typha latifolia the one that reached a more remarkable development. However, both species showed high resistance to the contaminated environment. These results showed higher removals than those reported in the literature with conventional Free Flow Vertical Constructed Wetlands (FFV-CWs), so PSV-CWs could be a suitable option to treat this type of effluent.

The research addresses the contamination of water resources in developing countries by landfill leachate and domestic wastewater discharges. It proposes treatment through Partially Saturated Vertical Constructed Wetlands (PSV-CWs), which, despite the limited information available, have been shown to be effective in removing pollutants in effluents with high concentrations. In addition to evaluating PSV-CWs, the study examines the impact of different types of vegetation on pollutant removal efficiency, concluding that PSV-CWs are a promising and viable option for the treatment of these effluents.

Influence of Bed Depth on the Development of Tropical Ornamental Plants in Subsurface Flow Treatment Wetlands for Municipal Wastewater Treatment: A Pilot-Scale Case.

The aim of this 2-year study was to evaluate the influence of bed depth (40 and 60 cm) on the development of tropical ornamental species (Alpinia purpurata, Heliconia latispatha and Strelitzia reginae) and on the removal of different contaminants such as chemical oxygen demand (COD), nitrate (N-NO3), ammonium (N-NH4), total nitrogen (TN), total phosphorus (TP), total suspended solids (TSS), total coliforms (TCs) and fecal coliforms (FCs), in horizontal subsurface flow constructed wetlands (HSSF-CWs) for municipal wastewater treatment. The results showed that the depth of 60 cm favored the removal of COD, with removal efficiencies of 94% for the three plant species. The depth of 40 cm was most effective for the removal of N-NH4 (80-90%). Regarding the removal of TN, the removals were similar for the different plants and depths (72-86%). The systems only achieved up to 60% removal of TCs and FCs. The depth of the CWs substrate and its saturation level influenced the development of ornamental vegetation, particularly flower production. For Heliconia latispatha, a bed depth level of 60 cm was more suitable, while for Alpinia purpurata 40 cm was better, and for Strelitzia reginae in both cases there was no flower production. The impact of bed depth on contaminant removal depends on the specific type of contaminant.

Variation in the Floral Morphology of Prosthechea karwinskii (Orchidaceae), a Mexican Endemic Orchid at Risk.

Prosthechea karwinskii is an orchid endemic to Mexico, threatened by the destruction of its habitat and the extraction of specimens to meet its demand for ornamental and religious use. Most of its populations, including the most locally abundant ones, are found in Oaxaca state. Variations in some floral traits have been observed in these populations. We implemented a morphometric analysis to assess their floral variation and identify the most significant characters in the morphological patterns of this orchid. Floral samples were collected from 17 populations of P. karwinskii in Oaxaca, as well as from specimens used as ornaments during Easter in an Oaxacan community (Zaachila), whose origin is unknown. Sampling of natural populations covered the environmental, geographic, and morphological variation of the species. We performed an analysis of variance (ANOVA), principal component analysis (PCA), canonical variate analysis (CVA), and cluster analysis, including 185 individuals and 45 variables (12 of them were discarded in the multivariate analyses due to high correlation). Characters of the column, lateral sepal, and labellum were most informative for the observed morphological patterns. Albarradas showed the greatest morphological differentiation, mainly due to the column. In general, individuals from the same locality tended to overlap more, especially the populations of Jaltianguis and Yahuiche, which were different from the geographically close population of Etla. Teposcolula presented the highest values in perianth characters, unlike Sola_Rancho Viejo. The specimens recovered from religious ornaments were morphologically more similar to those from Yanhuitlan and Etla. This morphometric analysis identified characters as potential taxonomic markers for P. karwinskii and related species, showing its potential to associate specimens of unknown origin with their probable geographical region. Our work encourages working on collaborative conservation strategies to ensure the long-term permanence of both the species and its traditional uses.

Feeding protocol for the Amazonian ornamental cichlid Apistogramma cacatuoides and a histological approach to the nutritional condition of larvae.

The aim of this study was to develop a feeding protocol for the larviculture of Apistogramma cacatuoides, using the histological approach to larval nutrition conditions. For this, three experiments were carried out. Experiment 1 was carried out in a randomized design to determine the optimal amount of Artemia nauplii (AN) per larva, and three treatments were evaluated: P1-feeding with 25 A. nauplii per larva (AN/L) during the first 5 days, followed by 50 AN/L from the 6th to the 10th day and 100 AN/L from the 11th to the 20th day; P2 and P3-37 and 50 AN/L during the first 5 days, 75 and 100 AN/L from the 6th to the 10th day and 150 and 200 AN/L from the 11th to the 20th day. Experiment 2 was carried out in a randomized design to determine the daily frequency of feeding and evaluated four feeding frequencies: F1-feeding only once a day (09:00); (F2)-feeding twice a day (09:00 and 17:00); F3-feeding three times a day (09:00, 11:30 and 17:00); and F4-feeding four times a day (09:00, 11: 30, 14:00 and 17:00). Experiment 3 lasted 40 days and was conducted in a randomized design to evaluate three periods for the beginning of the feeding transition: WE10:AN for 10 days, followed by 3 days of co-feeding and commercial feed until the end of the experimental period; WE15:AN for 15 days, followed by 3 days of co-feeding and commercial feed; WE20:AN for 20 days, followed by 3 days of co-feeding and commercial feed. The results of this study showed that, for the best development of the larvae, they should receive the feeding protocol 50-100-200 AN/L (P3) until the 20th day of exogenous feeding. From the 21st day, the transition to inert food should begin with 3 days of co-feeding, and feeding during larviculture should be carried out at a frequency of twice a day; this protocol provided a good nutritional status for the larvae, as shown by the histological approach.

Exploring floral morphoanatomy and embryology in wild populations of Chrysolaena flexuosa (Vernonia, Asteraceae): a contribution to understanding its ornamental potential.

Chrysolaena flexuosa (Sims.) H. Rob. is a South American species in the tribe Vernonieae, with potential ornamental value: it has attractive inflorescences, is suitable for pot cultivation, and its cypselae are useful for dried flower arrangements. Apart from studies on the growth dynamics of this species under cultivation, chromosome number, DNA content, ploidy level, size, pollen viability, and the characterization of phenotypic and genetic variability, it is noteworthy that other aspects regarding the floral architecture, reproductive mode, and gametophyte formation of C. flexuosa have not yet been studied. For this reason, our study encompasses a floral morphoanatomical survey and a comprehensive assessment of gametophyte development in the species. As a result of this study, we report new floral morphotypes, confirming that the morphological variability of the species might be greater than speculated. The morphoanatomy of the androecium and gynoecium and the male and female gametophyte developmental characteristics are uniform in all the populations studied despite the different ploidy levels. Chrysolaena flexuosa has five tetrasporangiate stamens of the dicotyledonous type of development; all the populations studied displayed a unilocular inferior ovary with a single anatropous, unitegumented, and tenuinucellar ovule. Given that all the embryo sacs observed were of the Polygonum-type development regardless of the ploidy level, we infer that the populations analyzed are fertile and undergo sexual reproduction. Our results not only contribute further research in the field of breeding systems and propagation of this species, but also promote the successful introduction of C. flexuosa to the plant ornamental market.

Characterization of Aeromonas Isolates from Ornamental Fish: Species, Virulence Genes, and Antimicrobial Susceptibility.

This study aimed to characterize 300 Aeromonas spp. strains isolated from 123 ornamental fish of 32 different species presenting with septicemia, skin lesions, and/or eye lesions. Within the 300 strains, 53.0% were identified as A. veronii, 41.3% as A. hydrophila, and 5.7% as A. caviae. Among the six virulence genes investigated, the most frequent were act (90.3%) and aer (79.3%). More than 50% of A. hydrophila strains were positive for all the studied genes. A total of 30 virulence profiles were identified, with the five main profiles identified comprising 75% of strains. Only five strains were negative for all genes and were identified as A. caviae and A. veronii. The antimicrobial susceptibility profile was performed for 234 strains, with sulfonamides presenting more than 50% of the resistance rates. Susceptibility was observed mainly for cephalosporins, aminoglycosides, chloramphenicol and piperacillin-tazobactam. Multidrug resistance was detected in 82.5% of the studied strains, including A. caviae with 100% multidrug resistance, and A. hydrophila with 90.9% multidrug resistance. The SE-AFLP analysis resulted in 66 genotypes of A. hydrophila, 118 genotypes of A. veronii, and 14 genotypes of A. caviae, demonstrating the greater heterogeneity of A. veronii and A. caviae. However, no direct correlation was observed between the genotypes and the strains' origins or virulence and resistance profiles.

Micropropagation of Guarianthe skinneri (Bateman) Dressler & W. E. Higging in Temporary Immersion Bioreactors.

Guarianthe skinneri (Bateman) Dressler & W. E. Higgins is an orchid valued for its ornamental characteristics. However, it is an orchid classified as threatened with extinction due to the illegal extraction from its natural habitat. In addition, its propagation through seed germination is very low, as is the case with most members of the family Orchidaceae. Its asexual propagation through pseudobulb separation is slow and produces a few propagules. For this reason, in vitro propagation techniques are an alternative to increase the number of plants obtained and thus be able to recover this valuable plant genetic resource. Temporary immersion systems (TIS) offer the advantage of mass-propagating plants for different purposes. This chapter describes a large-scale micropropagation protocol for Guarianthe skinneri using temporary immersion bioreactors (TIB).

Co-feeding period on the survival and growth of larvae (Botia lohachata) during the feeding transition / Co-feeding na sobrevivência e crescimento de larvas de botia yoyo Botia lohachata durante a transição alimentar

ABSTRACT: This study evaluated the effect of the co-feeding (C) period on the growth and survival of larvae of the yo-yo loach Botia lohachata ornamental species during the weaning period from live food (LF) to inert food (IF). The breeders were subjected to induced spawning using crude pituitary homogenate. A total of 2,400 larvae were used with an average initial weight of 0.39 ± 0.09 mg and an average total length of 4.08 ± 0.21 mm, from a single breeding pair. The live food was newly hatched nauplii of Artemia franciscana, and the inert diet was a commercial ration with 55% crude protein and two particle sizes (142-350 µm and 350-500 µm). As the first food, all larvae received A. franciscana during the first six days, except for the fasting (NC) and inert food (IF) groups. The following co-feeding strategies were employed: C4 = 4 days of co-feeding, C8 = 8 days of co-feeding, C12 = 12 days of co-feeding, and LF = Artemia during the entire experimental period. The longest period with live food promoted better larval growth rates. The specific growth rate of larvae fed live food during the entire period (LF) was higher than that of the other groups, followed by the co-feeding strategies. There was no effect (P > 0.01) on larval survival in the LF, C12, C8, and C4 groups, which ranged from 41% to 53% after 30 days. Treatment R resulted in high mortality and low growth, suggesting that live food should be the first food source for B. lohachata larvae.

RESUMO: Este estudo teve como objetivo avaliar o efeito do período de co-alimentação (C) no crescimento e sobrevivência de larvas da espécie ornamental Botia lohachata durante o período de desmame de alimento vivo (LF) para alimento inerte (IF). Os reprodutores foram submetidos à desova induzida com homogeneizado de hipófise. Foram utilizadas 2.400 larvas com peso inicial médio de 0,39 ± 0,09 mg e comprimento total médio de 4,08 ± 0,21 mm, de um único casal reprodutor. O alimento vivo foi náuplios recém-eclodidos de Artemia franciscana, e a dieta inerte foi uma ração comercial com 55% de proteína bruta e dois tamanhos de partículas (142-350 µm e 350-500 µm). Como primeiro alimento, todas as larvas receberam A. franciscana durante os primeiros seis dias, exceto os grupos jejum (NC) e alimento inerte (IF). As seguintes estratégias de co-alimentação foram empregadas: C4 = 4 dias de co-alimentação, C8 = 8 dias de co-alimentação, C12 = 12 dias de co-alimentação e LF = Artemia durante todo o período experimental. O maior período com alimento vivo promoveu melhores taxas de crescimento larval. A taxa de crescimento específico das larvas alimentadas com ração viva durante todo o período (LF) foi maior que a dos demais grupos, seguida das estratégias de co-alimentação. Não houve efeito (P > 0,01) na sobrevivência larval nos grupos LF, C12, C8 e C4, que variou de 41% a 53% após 30 dias. O tratamento IN resultou em alta mortalidade e baixo crescimento, sugerindo que o alimento vivo deve ser a primeira fonte de alimento para as larvas de B. lohachata.

Length-weight relationship for Potamotrygon wallacei (Carvalho, Rosa and Araújo, 2016) caught in the middle Negro River, Barcelos, Brazilian Amazon / Relação peso-comprimento de Potamotrygon wallacei (Carvalho, Rosa e Araújo, 2016) capturada no médio Rio Negro, Barcelos, Amazônia Brasileira

Abstract This research aimed to estimate the length-weight ratio (LWR) of the stingray Potamotrygon wallacei, known locally as the cururu, which was caught in streams and lakes in the middle Negro River region, Amazonas, Brazil. The stingrays were captured during the night (from 11 pm to 1 am) near the shores of streams and lakes, through active search using wooden canoes, head lanterns and scoop nets. The samplings were carried out in November 2017 (5 days), February (8 days), March (3 days) and April (2 days) of 2018, totaling 18 days of sampling. The total fresh weight was measured to 0.1 g of accuracy and the disc width to 0.1 cm accuracy. The parameters a and b of the equation W=a.DWb were estimated. This study provides new maximum length data for the cururu stingray.

Resumo Esta pesquisa teve como objetivo estimar a relação peso-comprimento (LWR) da arraia Potamotrygon wallacei, conhecida localmente como cururu, que foi capturada em igarapés e lagos na região do médio Rio Negro, Amazonas, Brasil. As arraias foram capturadas durante a noite (das 23h-1h) nas margens dos igarapés e lagos, por meio de busca ativa utilizando canoas de madeira, lanternas de cabeça e puçás. As coletas foram realizadas nos meses de novembro de 2017 (5 dias), fevereiro (8 dias), março (3 dias) e abril (2 dias) de 2018, totalizando 18 dias de amostragem. Foram medidos o peso fresco total com precisão de 0,01 g e a largura do disco com precisão de 0,1 cm. Os parâmetros a e b da equação W= a.DWb foram estimados. Este estudo prover novos dados de comprimento máximo para a arraia cururu.

Length-weight relationship for Potamotrygon wallacei (Carvalho, Rosa and Araújo, 2016) caught in the middle Negro River, Barcelos, Brazilian Amazon

Abstract This research aimed to estimate the length-weight ratio (LWR) of the stingray Potamotrygon wallacei, known locally as the cururu, which was caught in streams and lakes in the middle Negro River region, Amazonas, Brazil. The stingrays were captured during the night (from 11 pm to 1 am) near the shores of streams and lakes, through active search using wooden canoes, head lanterns and scoop nets. The samplings were carried out in November 2017 (5 days), February (8 days), March (3 days) and April (2 days) of 2018, totaling 18 days of sampling. The total fresh weight was measured to 0.1 g of accuracy and the disc width to 0.1 cm accuracy. The parameters a and b of the equation W=a.DWb were estimated. This study provides new maximum length data for the cururu stingray.

Resumo Esta pesquisa teve como objetivo estimar a relação peso-comprimento (LWR) da arraia Potamotrygon wallacei, conhecida localmente como cururu, que foi capturada em igarapés e lagos na região do médio Rio Negro, Amazonas, Brasil. As arraias foram capturadas durante a noite (das 23h-1h) nas margens dos igarapés e lagos, por meio de busca ativa utilizando canoas de madeira, lanternas de cabeça e puçás. As coletas foram realizadas nos meses de novembro de 2017 (5 dias), fevereiro (8 dias), março (3 dias) e abril (2 dias) de 2018, totalizando 18 dias de amostragem. Foram medidos o peso fresco total com precisão de 0,01 g e a largura do disco com precisão de 0,1 cm. Os parâmetros a e b da equação W= a.DWb foram estimados. Este estudo prover novos dados de comprimento máximo para a arraia cururu.

Efecto del nitrato de plata (AgNO3) en el crecimiento in vitro de brotes de Anthurium magnificum Linden

Anthurium magnificum Linden es una importante planta ornamental perteneciente a la familia Araceae. Esta especie es muy demandada como planta de maceta para la decoración de interiores y jardines, así como follaje de corte. Los métodos de propagación tradicionales en esta especie presentan ciertas desventajas por lo que no permiten satisfacer la creciente demanda. Estas limitantes convierten al cultivo de tejidos vegetales en el método más eficiente para propagar plantas de Anthurium magnificum a un ritmo más rápido que los métodos de propagación convencionales. En el cultivo de tejidos vegetales el nitrato de plata adicionado a los medios de cultivo actúa como un inhibidor de la acción del etileno y desempeña un papel crucial en la regulación de procesos fisiológicos esenciales de las plantas. El presente trabajo tuvo como objetivo determinar el efecto de diferentes concentraciones de nitrato de plata en el crecimiento in vitro de brotes de Anthurium magnificum. Para ello, se cultivaron brotes in vitro de Anthurium magnificum en medios de cultivo enriquecidos con diferentes concentraciones de nitrato de plata. Los resultados mostraron que los mayores valores en las variables altura de la planta, número de raíces y longitud de las raíces se obtuvieron en un medio de cultivo con 1,0 mg L-1 de nitrato de plata. En la variable área foliar de la planta los mayores valores se presentaron en los medios de cultivo que contenían nitrato de plata independientemente de la concentración empleada.

Anthurium magnificum Linden is an important ornamental plant belonging to the Araceae family. This species is in great demand as a pot plant for interior and garden decoration, as well as cut foliage. The traditional propagation methods in this species have certain disadvantages, which is why they cannot satisfy the growing demand. These limitations make plant tissue culture the most efficient method to propagate Anthurium magnificum plants at a faster rate than conventional propagation methods. In the culture of plant tissues, silver nitrate added to the culture media acts as an inhibitor of the action of ethylene and plays a crucial role in the regulation of essential physiological processes in plants. The objective of this work was to determine the effect of different concentrations of silver nitrate on the in vitro growth of Anthurium magnificum shoots. For this, in vitro shoots of Anthurium magnificum were cultivated in culture media enriched with different concentrations of silver nitrate. The results showed that the highest values in the variables plant height, number of roots and length of the roots were obtained in a culture medium with 1.0 mg L-1 of silver nitrate. In the variable leaf area of the plant, the highest values were presented in the culture media that contained silver nitrate, regardless of the concentration used.

Morpho-anatomy of in vitro germination and cryopreservation of the orchid Cattleya crispa (Orchidaceae Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae) / Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae)

Introduction: Cattleya crispa is an ornamental epiphytic orchid with geographic distribution restricted to the Brazilian Atlantic Forest. Due to predatory extractivism and human-induced habitat loss, this species appears on the Red List of Brazilian Flora. Objective: To characterize morpho-anatomical aspects regarding germination and post-seminal development from C. crispa seeds; as well as studying the effect of cryopreservation on these seeds. Methods: We used light microscopy and electron microscopy to describe the microstructure of a 100 ripe seeds. We evaluated seed viability, seed germination, survival rate and protocorm weight in cryopreserved and non-cryopreserved material, with four replicas per treatment using 20 mg of plant material. Results: The seeds are fusiform, whitish yellow with a length from 700 to 900 µm and a water content of 5 %. Germination began seven days after sowing, the formation of the globular protocorm at 30 days and the formation of the seedling occurred 150 days. The persistent seed coat can compress the protocorm and cause it to collapse. The cryopreserved seeds presented 87.15 % viability, 78.32 % germination, 8.48 % survival and protocorms with 104.27 mg five months after sowing. Data wasn't different to non-cryopreserved seeds. Conclusions: The cryocapability of the seeds shows that cryopreservation can be used for long-term conservation. The results of this work contribute to the overall biology of C. crispa and to the propagation and storage of genetic material for conservation purposes.

Introducción: Cattleya crispa es una orquídea epífita ornamental con distribución geográfica restringida a la Mata Atlántica brasileña. Debido al extractivismo depredador y a la pérdida de hábitat inducida por el hombre, esta especie aparece en la Lista Roja de la Flora Brasileña. Objetivo: Caracterizar aspectos morfoanatómicos de la germinación y desarrollo inicial de semillas de C. crispa; así como estudiar el efecto de la criopreservación de estas semillas. Métodos: Utilizamos microscopía óptica, microscopía electrónica de barrido y microscopía electrónica de transmisión para describir la microestructura en 100 semillas maduras. Evaluamos la viabilidad de la semilla, la germinación de la semilla, la tasa de supervivencia y el peso de los protocormos en el material criopreservado y no criopreservado, con cuatro réplicas por tratamiento de 20 mg de material vegetal. Resultados: Las semillas son fusiformes, amarillo blanquecinas, con una longitud de 700 a 900 µm y un contenido de agua del 5 %. La germinación comenzó siete días después de la siembra, la formación del protocormo globular a los 30 días y la formación de la plántula a los 150 días. La cubierta de semilla persistente puede comprimir el protocormo y provocar su colapso. Las semillas criopreservadas presentaron 87.15 % de viabilidad, 78.32 % de germinación, 8.48 % de supervivencia y protocormos con 104.27 mg a los cinco meses de la siembra. Los datos no fueron diferentes a las semillas no criopreservadas. Conclusiones: La capacidad criogénica de las semillas muestra que la crioconservación puede utilizarse para la conservación a largo plazo. Los resultados de este trabajo contribuyen a la biología general de C. crispa y a la propagación y almacenamiento de material genético con fines de conservación.

On a new colorful species of Moenkhausia (Characiformes: Characidae) from the upper rio Madeira basin at the Chapada dos Parecis, Brazil, with comments on its conservation and putative biogeographic history.

A new species, Moenkhausia iris, is described from the upper rio Madeira basin, Mato Grosso State, Brazil. The new species is unique among congeners by its color in life, which is the dorsal half of body blue or green and ventral half red, more intense above the anal fin. Additionally, it can be distinguished by having a single, vertically elongated humeral spot, a lateral surface of body, caudal peduncle, and anal fin without distinct blotches or oblique marks and 18-19 branched anal-fin rays. The new species is endemic from a small tributary of the Rio Guaporé, rio Madeira drainage at Chapada dos Parecis. Its putatively close relatives are endemic to the upper rio Juruena located at the same shield. Both headwaters are very close to each other, sharing other restricted range species, and their hydrogeological topography indicates the upper rio Juruena stretch was captured by the rio Guaporé through a headwater capture event. Moenkhausia iris is much appreciated in the international aquarium trade, as well as many other tetras endemic from the same area. Exportation is based on the capture of wild individuals so sustainable exploitation is necessary for its conservation.

Diversity of Colletotrichum species associated with torch ginger anthracnose.

Anthracnose caused by Colletotrichum species is one of the most important diseases of torch ginger. The disease leads to loss of aesthetic and commercial value of torch ginger stems. This study aimed to characterize Colletotrichum species associated with torch ginger anthracnose in the production areas of Pernambuco and Ceará. A total of 48 Colletotrichum isolates were identified using molecular techniques. Pathogenicity tests were performed on torch ginger with representative isolates. Phylogenetic analyses based on seven loci-DNA lyase (APN2), intergenic spacer between DNA lyase and the mating-type locus MAT1-2-1 (APN2/MAT-IGS), calmodulin (CAL), intergenic spacer between glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and a hypothetical protein (GAP2-IGS), glutamine synthetase (GS), and ß-tubulin (TUB2)-revealed that they belong to five known Colletotrichum species, namely, C. chrysophilum, C. fructicola, C. siamense, C. theobromicola, and C. tropicale, and three newly discovered species, described here as C. atlanticum, C. floscerae, and C. zingibericola. Of these, C. atlanticum was the most dominant. Pathogenicity assays showed that all isolates were pathogenic to torch ginger bracts. All species are reported for the first time associated with torch ginger in Brazil. The present study contributes to the current understanding of the diversity of Colletotrichum species associated with anthracnose on torch ginger and demonstrates the importance of accurate species identification for effective disease management strategies.

Phylogeography and population genetic structure of the cardinal tetra (Paracheirodon axelrodi) in the Orinoco basin and Negro River (Amazon basin): evaluating connectivity and historical patterns of diversification.

The Neotropics contain one of the most diverse assemblages of freshwater fishes worldwide. Part of this diversity is shared between the Orinoco and Amazon basins. These basins have been separated for a long time due to the Vaupes Arch, rising between 10-11 Ma. Today, there is only one permanent connection between the Orinoco and Negro (Amazon) basins, known as the Casiquiare Canal. However, alternative corridors allowing fish dispersion between both basins have been proposed. The cardinal tetra (Paracheirodon axelrodi), the most important fish in the ornamental world market, is distributed in both basins. Here we investigated P. axelrodi phylogeography, population structure, and potential routes of migration and connectivity between the two basins. A total of 468 bp of the mitochondrial gene (COI), 555 bp of the nuclear gene fragment (MYH6), and eight microsatellite loci were analyzed. As a result, we found two major genetic clusters as the most likely scenario (K = 2), but they were not discreetly distributed between basins. A gradient of genetic admixture was observed in Cucui and São Gabriel da Cachoeira, between the upper Negro River and the upper Orinoco. Samples from the middle-lower Negro River were highly structured. Cucui (Negro basin) was more similar to the Orinoco than to the rest of the Negro basin populations. However, substructure was also observed by the discriminant analysis, fixation indices and other hierarchichal structure analyses (K = 3 - 6), showing three major geographic clusters: Orinoco, Cucui, and the remaining Negro basin. Unidirectional migration patterns were detected between basins: via Cucui toward Orinoco and via the remaining of the Negro basin toward Orinoco. Results from the Relaxed Random Walk analysis support a very recent origin of this species in the headwater Orinoco basin (Western Guiana Shield, at late Pleistocene) with a later rapid colonization of the remaining Orinoco basin and almost simultaneously the Negro River via Cucui, between 0.115 until about 0.001 Ma. Historical biogeography and population genetic patterns observed here for Cardinal tetra, seem to be better explained by river capture, physical, or ecological barriers than due to the geographic distance.

Karyotypic and Molecular Analysis of Pterygoplichthys pardalis (Castelnau 1855) from the Lower Amazon River.

Pterygoplichthys pardalis is an armored catfish native to South America and an important resource for the ornamental fish industry. Recently, several exotic populations have been introduced into rivers on five continents. Despite its commercial and environmental importance, P. pardalis is poorly studied from a genetic perspective. In this study, we analyzed the karyotype of P. pardalis from the Amazon River and molecular variations in the mitochondrial gene Cytochrome oxidase I (COI) between native and exotic populations. The karyotype presented diploid number 2n = 52 and NF = 100 without cytogenetic variation between males and females. Nucleolus organizer regions (Ag-NOR) in the distal region of the long arm of pair 12 coincided with the 18S hybridization signal, whereas 5S was syntenic to this chromosome but localized in the short arm. The constitutive heterochromatin was restricted in the distal regions of pairs 4, 12, 25, and 26. Telomeric probes showed only distal hybridization signals. The karyotype of P. pardalis diverged from that of its congeners, and COI molecular variation revealed four haplotypes. The Philippine population revealed the greatest diversity with three haplotypes, while haplotype H1 was the most abundant and observed in both native and exotic populations. This new genetic data contributes to species management and provides useful information from an aquaculture perspective.