RESUMEN
Post-thawing cryoinjuries in boar spermatozoa due to oxidative stress may be reduced by adding nanoencapsulated antioxidants to freezing extenders. This study evaluated post-thawing kinetics, structural and biochemical functions of boar spermatozoa frozen with extenders including resveratrol and vitamin E loaded into polymeric nanocapsules. Resveratrol was added at 0 (control), 5, 10, 20, 40 and 80 µg/ml, whereas Vitamin E was added at 0 (control), 50, 100, 200 and 400 µg/ml. Both antioxidants were tested in free and nanoencapsulated presentations. In contact with empty nanocapsules, some sperm kinetics parameters were impaired compared to the control (p < .05), whereas lipoperoxidation declined (p < .05). With inclusion of 40 µg/ml nanoencapsulated resveratrol, some sperm kinetics parameters were improved (p < .01), but sperm motility, structural and biochemical functions did not differ from the control (p > .05). No improvement in sperm quality occurred with inclusion of vitamin E, although sperm kinetics with 400 µg/ml nanoencapsulated vitamin E was reduced compared to the control (p < .01). Inclusion of 40 µg/ml nanoencapsulated resveratrol benefitted boar sperm kinetics after thawing, but no improvement resulted from inclusion of vitamin E.
Asunto(s)
Preservación de Semen , Animales , Antioxidantes/farmacología , Criopreservación , Crioprotectores/farmacología , Congelación , Masculino , Preservación de Semen/veterinaria , Motilidad Espermática , Espermatozoides , PorcinosRESUMEN
Sea anemones of the genus Bunodosoma possess along their body column, longitudinally arranged brown-colored vesicles. We have shown that in B. cangicum, these warty structures contain a mixture of potent toxins. This work highlights the neuro-inhibitory effects exhibited by two decapod crustacean species exposed to the extracts from these vesicles. For this, we use the unrefined toxin in doses, exposure times, and different exposure pathways. The findings show that at least one neuro-inhibitory compound is present and remains active regardless of the exposure method or dose tested. This toxin affects neuro-motor pathways but not neuro-sensory pathways. Shrimp exposed to toxin could continue to perceive and track food pellets but could not secure and consume their ration. Of six anatomical reflexes tested under the toxin's influence, voluntary movements of the mouthparts were impacted most commonly. Interestingly, all subject animals recovered from the toxin exposure within 2 h. Finally, we propose Reflexive Action Analysis (RAMP) as a tool to evaluate the potency of other neurotoxic or neuro-inhibitory compounds in crustacea. This work is the first to show the neuro-inhibitory activity of extracts from these sea anemone columnar vesicle structures and the first to evaluate these effects using RAMP reflex analysis.
Asunto(s)
Conducta Animal/efectos de los fármacos , Braquiuros/efectos de los fármacos , Venenos de Cnidarios/toxicidad , Palaemonidae/efectos de los fármacos , Anémonas de Mar/química , Animales , Braquiuros/fisiología , Agua Dulce/química , Monitorización Neurofisiológica/métodos , Palaemonidae/fisiología , Anémonas de Mar/metabolismo , Alimentos MarinosRESUMEN
Physical methods such as electroporation have been used to improve the DNA uptake efficiency of sperm cells. This study aims to develop an efficient capillary-type electroporation method for incorporation of exogenous DNA into bovine cryopreserved sperm cells with minimal detrimental effects for later use in SMGT. Electroporation of the samples was performed in 2 different groups (with 1 µg of DNA and without DNA transfection) and under five different voltages: 500 V, 600 V, 700 V, 800 V and 900 V. Non-electroporated sperm cells (with and without DNA) were used as control. Kinetics parameters were determined using computer assisted semen analyses, whereas membrane integrity, fluidity, mitochondrial function and DNA uptake were evaluated by flow cytometry. Results revealed that all tested voltages reduced electroporated sperm motility (P < 0.05) when compared to the control (non-electroporated cells). Mitochondrial function results showed no statistical difference among groups. Similarly, groups electroporated with lower (500 V, 600 V and 700 V) voltages showed no difference in cell membrane integrity and fluidity. Groups electroporated at higher voltages (800 V and 900 V) demonstrated negative effects in cells membrane integrity when compared to other groups and control. Also, all electroporated groups demonstrated significant higher percentages of transfected sperm cells when compared to the control group (P < 0.05). Under the recommendation of using voltages up to 600 V, this method represents a safe and efficient alternative for electroporation of bovine spermatozoa.
Asunto(s)
Electroporación/métodos , Preservación de Semen , Espermatozoides/fisiología , Animales , Bovinos , Criopreservación , Masculino , Análisis de Semen , Motilidad Espermática/fisiologíaRESUMEN
Roundup® formulations are the most consumed glyphosate-based herbicides in the world. When applied, they may reach water bodies and exert toxicity toward non-target species. This study evaluated and compared the effects of different variations of Roundup on biochemical biomarkers as oxidative parameters and Acethylchorinesterase (AChE) activity, and sperm quality of the livebearing Jenynsia multidentata. Fish were acutely (96 h) exposed to Roundup Original® (RO), Roundup Transorb® (RT) and Roundup WG® (RWG) at 0.0, 0.5, 1 and 5 mg L-1 of nominal glyphosate. The highest mortality (60%) was observed for fish exposed to RT at the highest concentration tested and at 0.5 mg L-1 non-mortality was observed, so this concentration was chosen for the experiments. Fish exposed to RO and RT (24 and 96 h) presented a state of oxidative imbalance, which caused lipid peroxidation (LPO) in their livers. Oxidative stress was more severe in RO treatment, which may be resulted in the highest hepathosomatic index at 96 h. However, fish exposed to RT presented a marked inhibition of AChE activity from membrane cells of muscle and brain tissues. Sperm quality was investigated in livebearing exposed (24 and 96 h) to the three formulations. Spermatozoa motility and concentration were affected by all formulations. Overall, Roundup formulations are harmful to the fish J. multidentata at 0.5 mg L-1 of glyphosate; however, mechanisms and potential of toxicity are different between formulations. The J. multidentata also represents a sensitive species and a good regional bio-monitor.
Asunto(s)
Biomarcadores/química , Ciprinodontiformes , Herbicidas/toxicidad , Espermatozoides/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Animales , Glicina/análogos & derivados , Glicina/toxicidad , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Masculino , Estrés Oxidativo/efectos de los fármacos , GlifosatoRESUMEN
The objective of this study was to assess the placental vasculature and microcotyledons in pregnant mares with chronic laminitis. Twenty-six pregnant mares were enrolled in the study, 13 had chronic laminitis (Laminitis Group) and 13 were healthy mares (Healthy Group). Arterial systolic pressure and heart rate were measured in the last 30 days of gestation. After foaling, the fetal membranes were grossly evaluated and samples were harvested for histopathologic examination. All mares had digitalized images taken from chorioallantois for histomorphometry analyses (software-NIH ImageJ). Images were assessed for: (i) arterioles from the allantoic region: total and lumen vascular diameter and vascular wall thickness; (ii) microcotiledonary and capillary area/field. Mares in the Laminitis Group showed hypertension, shorter gestational length, lower placental weight and lower birthweight (p < 0.05) foal in comparison with mares in the Healthy Group. Laminitis mares had a reduction of vascular lumen diameters in the uterine body and pregnant horn (p < 0.05), vascular wall thickening in the pregnant horn (p < 0.05) and smaller capillary area/field in the microcotyledons of uterine body and pregnant horn (p < 0.05). In conclusion, pregnant mares with chronic laminitis presented signs of hypertension syndrome, and vascular abnormalitities in placental vessels such as reduction in the vascular lumen and capillary area in the microcotyledones, and thickening of the vascular wall. Foals born from mares with chronic laminitis showed lower birth weight and shorter gestation lengths.
Asunto(s)
Enfermedades del Pie/veterinaria , Pezuñas y Garras/patología , Enfermedades de los Caballos/patología , Caballos/embriología , Placenta/irrigación sanguínea , Animales , Femenino , Enfermedades del Pie/complicaciones , Placenta/patología , EmbarazoRESUMEN
We provide ultrastructural and cytological evidence that the tentacles of the sea anemone Bunodosoma cangicum does not contain cytotoxic venom. However, we show that the stimulated secretion of an apparent mixture of biomolecules containing polypeptides from the columnar vesicles of Bunodosoma cangicum is apparently a potent inducer of apoptosis in the zebrafish cell line, ZF-L. Microscopic fluorescence, cell morphology and flow cytometric assays confirm the apoptotic activity. Crude vesicle venom was partially purified by size exclusion chromatography. PAGE analysis shows that this venom contains low weight polypeptides but no measurable protein. The apoptotic activity is heat labile, and the observed peptides concurrent with this activity have a molecular weight of approximately 2000 Da. This manuscript is the first report of biologically active molecules and peptides associated with columnar vesicles of anemones, and the first to confirm that the tentacles of B. cangicum do not contain cytotoxic venom, and express spirocytes exclusively.
Asunto(s)
Apoptosis/efectos de los fármacos , Venenos de Cnidarios/farmacología , Péptidos/metabolismo , Anémonas de Mar/metabolismo , Animales , Línea Celular , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Microscopía Electrónica de Rastreo , Pez CebraRESUMEN
Silver (Ag), cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) concentrations were analyzed in tissues of juvenile green sea turtles (Chelonia mydas) found stranded along the southern Atlantic coast in Brazil. Green sea turtles were collected (n=29), measured (curved carapace length: CCL) and had their muscle, liver, and kidney dissected for metal concentration measurements. Sex was identified in 18 individuals (10 females and 8 males) through gonad histology. No gender differences in CCL and tissue metal concentrations were observed. In the muscle, there was a negative correlation between CCL and Cd and Cu concentrations. Metal concentrations were lower in the muscle than in the liver and kidney. Zn concentration in the muscle was the highest of all metals analyzed (16.6 mg/kg). The kidney showed the highest concentrations of Pb, Cd and Zn (5.4, 28.3 and 54.3 mg/kg, respectively), while the liver had the highest values of Ag and Cu (0.8 and 100.9 mg/kg, respectively). Tissue Ag, Zn and Cd concentrations were similar to those found in green sea turtles from other regions while Cu and Pb values were elevated, likely due to the metal-rich water and sediment reported in the collection area. In the liver and kidney, concentrations of non-essential (Ag, Cd and Pb) and essential (Cu or Zn) metals were positively correlated, likely due to an induced metallothionein synthesis to protect tissue against the toxic effect of metals. This is the first study to report and correlate the concentrations of essential and non-essential metals in tissues of green sea turtles in the Brazilian southern Atlantic coast, an important feeding and developing area for this turtle species.