RESUMEN
This study increases the known biodiversity of cnidarian parasites in neotropical bryconid fishes. Two novel Myxobolus species are described based on morphology, ultrastructure and small subunit ribosomal DNA (ssrDNA) sequencing: Myxobolus vetuschicanus n. sp. infecting fins of Salminus franciscanus and Myxobolus mineirus n. sp. infecting the mesentery of Brycon orthotaenia from the São Francisco River basin, Minas Gerais State, Brazil. Ultrastructural analysis of the two species revealed an asynchronous sporogenesis process, with germinative cells and young developmental stages of myxospores in the periphery of the plasmodia. In M. vetuschicanus n. sp., the plasmodia were surrounded by a layer of fibroblasts and in M. mineirus n. sp., the plasmodial membrane had direct contact with the host tissue. The phylogenetic analysis based on the ssrDNA of Henneguya/Myxobolus species showed that the two novel Myxobolus species grouped in subclades together with other parasite species of bryconid fishes.
Asunto(s)
Biodiversidad , Characiformes/parasitología , Enfermedades de los Peces/parasitología , Myxobolus/aislamiento & purificación , Enfermedades Parasitarias en Animales/parasitología , Aletas de Animales/parasitología , Animales , Brasil , ADN Ribosómico , Branquias/parasitología , Myxobolus/clasificación , Filogenia , Subunidades Ribosómicas Pequeñas de Eucariotas , Ríos/parasitologíaRESUMEN
Nitric oxide (NO) is an important effector molecule which is involved in a myriad of biological processes, including immune responses against pathogens such as parasites, virus and bacteria. During the inflammatory processes in vertebrates, NO is produced by the inducible nitric oxide synthase (iNOS) enzyme in practically all nucleated cells to suppress or kill intracellular pathogens. The aim of the present study was to characterize the full coding region of the iNOS gene of pacu (Piaractus mesopotamicus), an economically and ecologically important South American fish species, and to analyze mRNA expression levels following intraperitoneal infection with the pathogenic bacterium Aeromonas dhakensis by means of quantitative real time PCR (qPCR). The results showed that the pacu iNOS transcript is 3237 bp in length, encoding a putative protein composed of 1078 amino acid residues. The amino acid sequence showed similarities ranging from 69.03% to 94.34% with other teleost fish and 57.70% with the human iNOS, with all characteristic domains and cofactor binding sites of the enzyme detected. Phylogenetic analysis showed that the iNOS from the red-bellied piranha, another South American characiform, was the closest related sequence to the pacu iNOS. iNOS transcripts were constitutively detected in the liver, spleen and head kidney, and there was a significant upregulation in the liver and spleen at 12, 24 and 48â¯h after infection with A. dhakensis. No significant variations were observed in the head kidney during the periods analyzed. These results show that iNOS expression was induced by A. dhakensis infection and suggest that this enzyme may be involved in the response to this bacterium in pacu.
Asunto(s)
Characiformes/genética , Characiformes/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/inmunología , Inmunidad Adaptativa , Aeromonas/inmunología , Secuencia de Aminoácidos , Animales , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica , Infecciones por Bacterias Gramnegativas/inmunología , Óxido Nítrico Sintasa de Tipo II/química , Filogenia , Distribución Aleatoria , Alineación de Secuencia/veterinariaRESUMEN
The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study, we demonstrated that IVM leads to accumulation of neutral lipids associated with differential levels of the mono-, di- and triacylglycerols in both cumulus cells and oocytes. We observed that in vitro-matured oocytes exhibited decreased glutathione and reactive oxygen species levels and a lower ATP/ADP ratio when compared to in vivo-matured oocytes, with no significant differences in metabolism and stress-related mRNA or miRNA levels. Moreover, in addition to an increase in lipids in in vitro-matured cumulus cells, fatty acid synthesis and accumulation as well as glycolysis pathway genes were upregulated, whereas those affiliated with the ß-oxidation pathway were decreased. Our gene expression data in cumulus cells suggest the disruption of endoplasmic reticulum stress, apoptosis and cellular stress response pathways during IVM. Furthermore, a total of 19 miRNAs were significantly altered by the maturation process in cumulus cells. These results indicate some new negative influences of the in vitro system in cumulus-oocyte complexes, demonstrating the occurrence of functional disruption in lipid metabolism and stress pathways and showing evidences suggesting the occurrence of altered mitochondrial activity and energy metabolism during IVM, with a massive dysregulation of the corresponding transcripts in the surrounding cumulus cells.
Asunto(s)
Células del Cúmulo/metabolismo , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oocitos/metabolismo , Estrés Oxidativo , Animales , Bovinos , Células Cultivadas , Células del Cúmulo/citología , Metabolismo Energético , Femenino , Oocitos/citología , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The development of biomaterials to enable application of antimicrobial peptides represents a strategy of high and current interest. In this study, a bioparticle was produced by the complexation between an antimicrobial polypeptide and the biocompatible and biodegradable polysaccharides chitosan-N-arginine and alginate, giving rise to a colloidal polyelectrolytic complex of pH-responsive properties. The inclusion of the polypeptide in the bioparticle structure largely increases the binding sites of complexation during the bioparticles production, leading to its effective incorporation. After lyophilization, detailed evaluation of colloidal structure of redispersed bioparticles evidenced nano or microparticles with size, polydispersity and zeta potential dependent on pH and ionic strength, and the dependence was not withdrawn with the polypeptide inclusion. Significant increase of pore edge tension in giant vesicles evidenced effective interaction of the polypeptide-bioparticle with lipid model membrane. Antibacterial activity against Aeromonas dhakensis was effective at 0.1% and equal for the isolated polypeptide and the same complexed in bioparticle, which opens perspectives to the composite material as an applicable antibacterial system.
RESUMEN
This paper presents the morphological, histological and ultrastructural characteristics of Myxobolus oliveirai sp. nov., a parasite of the gill filaments in Brycon hilarii from the Brazilian Pantanal. Out of 216 B. hilariispecimens examined (126 wild and 90 cultivated), 38.1% of wild specimens (n = 48) were infected. The parasites form elongated plasmodia primarily in the tip of gill filaments, reaching about 3 mm in length. A thorough comparison with all the Myxobolus species described from South American hosts, as well as nearly all the Myxobolus species described so far is provided. Partial sequencing of the 18S rDNA gene revealed a total of 1,527 bp. The Myxobolus species parasite of B. hilarii did not match any of the Myxozoa available in GenBank. In the phylogenetic analysis, M. oliveirai sp. nov. composed a monophyletic group with eight other species: five species of Myxobolus parasites of mugilid fishes, two parasites of pangasiid and one of centrarchid. Infection prevalence values of the parasite revealed no significant differences between wet and dry seasons or between males and females. The importance of the infection to the farming of the host species is emphasized.
Asunto(s)
Enfermedades de los Peces/parasitología , Peces/parasitología , Myxobolus/genética , Myxobolus/ultraestructura , Animales , Brasil/epidemiología , ADN Protozoario/análisis , ADN Ribosómico/análisis , Femenino , Enfermedades de los Peces/epidemiología , Branquias/parasitología , Masculino , Microscopía Electrónica de Transmisión , Myxobolus/aislamiento & purificación , Filogenia , Prevalencia , Estaciones del AñoRESUMEN
The grazing of Zebu cattle in poor-quality tropical pastures during the dry season has an increased environmental impact and cost of production. The use of condensed tannins (CT) as a natural feed additive to modulate ruminal archaea can mitigate the methane emissions from cattle in tropical systems. We investigated the effects of CT on in vivo methane emissions and rumen microbiota ecology in beef cattle. Batch experiments were also conducted to evaluate the impact of dietary CT on the biogas production from beef cattle manure. Six adult rumen-cannulated Nellore cattle were used in a double 3 × 3 Latin square design. Treatments consisted of three diets containing either a 0%, 1.25% or 2.5% CT additive from Acacia mimosa extract. The experimental period consisted of 63 days, and methane production was measured using the sulfur hexafluoride (SF6) technique from Day 16 to 21 of each feeding period. Adding Acacia extract to the diets reduced daily methane emissions per animal. Methane suppression occurred more by reduction of intake than by the direct effect on methanogenic archaea. We verified that CT directly suppresses archaea rumen communities and increases total rumen bacteria. Our study indicates that CT benefit rumen Fibrobactersuccinogenes and Ruminoccous flavefaciens populations and have no negative effect on biogas production from cattle manure. Acacia extract as a feed additive has promising potential as part of an overall nutritional strategy to reduce the methanogenesis from Zebu beef cattle in tropical systems.
Asunto(s)
Archaea/efectos de los fármacos , Biocombustibles/análisis , Bovinos/metabolismo , Metano/biosíntesis , Proantocianidinas/metabolismo , Rumen/metabolismo , Alimentación Animal/análisis , Animales , Archaea/metabolismo , Brasil , Dieta/veterinaria , Suplementos Dietéticos/análisis , Masculino , Rumen/microbiología , Clima TropicalRESUMEN
Ranavirus is a double-stranded DNA virus associated with amphibian, fish and reptile die-offs worldwide. International trade of live animals farmed for human consumption, such as the American bullfrog (Lithobates catesbeianus), plays a key role in spreading the pathogen. In Brazil, ranavirus has only been reported in captive bullfrog farms. We found infected tadpoles of both native species and the American bullfrog in the wild, and a case of mass mortality of amphibians and fish potentially associated with ranavirus. Dead animals presented skin ulcerations, hemorrhages, and edemas. We also found an overall prevalence of 37% of the amphibian chytrid in the area, and two bullfrog tadpoles were co-infected with both pathogens. We suggest that the interaction between the two pathogens should be investigated to improve global conservation of ectothermic vertebrates.
Asunto(s)
Infecciones por Virus ADN/veterinaria , Rana catesbeiana/virología , Ranavirus/aislamiento & purificación , Animales , Brasil/epidemiología , Infecciones por Virus ADN/epidemiología , Infecciones por Virus ADN/virología , Larva/virologíaRESUMEN
This paper presents the morphological, histological and ultrastructural characteristics of Myxobolus oliveirai sp. nov., a parasite of the gill filaments in Brycon hilarii from the Brazilian Pantanal. Out of 216 B. hilariispecimens examined (126 wild and 90 cultivated), 38.1 percent of wild specimens (n = 48) were infected. The parasites form elongated plasmodia primarily in the tip of gill filaments, reaching about 3 mm in length. A thorough comparison with all the Myxobolus species described from South American hosts, as well as nearly all the Myxobolus species described so far is provided. Partial sequencing of the 18S rDNA gene revealed a total of 1,527 bp. The Myxobolus species parasite of B. hilarii did not match any of the Myxozoa available in GenBank. In the phylogenetic analysis, M. oliveirai sp. nov. composed a monophyletic group with eight other species: five species of Myxobolus parasites of mugilid fishes, two parasites of pangasiid and one of centrarchid. Infection prevalence values of the parasite revealed no significant differences between wet and dry seasons or between males and females. The importance of the infection to the farming of the host species is emphasized.