Nanocarried antioxidants in freezing extenders for boar spermatozoa.

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.

The effects of vesicle toxin from the sea anemone Bunodosoma cangicum on the behavior of a freshwater shrimp, Palaemon argentinus, and shore crab, Neohelice granulata.

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.

Triphenyltin exposition induces spermatic parameter alters of Calomys laucha species.

The present study aims to evaluate the influence of triphenyltin (TPT) exposure on reproductive physiology on Calomys laucha species, since this species inhabits regions susceptible to exposure to this contaminant. Animals exposed to the highest dose (10.0 mg/kg) presented signs of severe intoxication in only 7 days of exposure, demonstrating a higher sensitivity of this species to triphenyltin. The 10.0 mg TPT/kg dose was analyzed separately for short-term exposure and results suggest that exposure to this dose was severely detrimental to sperm activity. Among the main results obtained in the evaluation of sperm kinetics, a reduction in total motility was observed from the 0.5 mg TPT/kg group, accentuated according to the increase in the doses of TPT. In progressive motility, there was a decrease from the dose of 0.5 mg TPT/kg and maintained the plateau until the dose of 5.0 mg TPT/kg. It was also observed an increase in the distances and velocities average path, rectilinear and curvilinear in doses of 2.5 and 5.0 mg/kg. From the flow cytometry, evaluation a decrease in mitochondrial functionality was observed as the dose increased. Increased membrane fluidity was also observed from the 5.0 mg TPT/kg dose and the acrosome reaction presented higher values at doses of 0.5 and 5.0 mg TPT/kg. We can conclude that TPT causes impairment of the sperm activity, reducing it in individuals exposed in the adult phase.

Voltages up to 600V did not affect cryopreserved bovine spermatozoa on capillary-type electroporation.

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.

Effects of Roundup formulations on biochemical biomarkers and male sperm quality of the livebearing Jenynsia multidentata.

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.

Histomorphometry of the placental vasculature and microcotyledons in Thoroughbred mares with chronic laminitis.

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.

Who knows not where an anemone does wear his sting? Could polypeptides released from the columnar vesicles of Bunodosoma cangicum induce apoptosis in the ZF-L cell line?

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.

Concentrations and distributions of metals in tissues of stranded green sea turtles (Chelonia mydas) from the southern Atlantic coast of Brazil.

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.

Toxic effects of the herbicide Roundup in the guppy Poecilia vivipara acclimated to fresh water.

Although it is believed that glyphosate-based herbicides are relatively nontoxic to humans, its broad use in agriculture and consequent contamination of aquatic systems is a concern. In the present study, reproductive (sperm quality) and biochemical parameters (acetylcholinesterase and glutathione S-transferase activity, lipoperoxidation, and antioxidant capacity against peroxyl radicals) were evaluated in adult guppies (Poecilia vivipara) acclimated to fresh water and exposed (96 h) to environmentally realistic concentrations of glyphosate (130 and 700 µg L(-1)) as the commercial formulation Roundup. Male guppies exposed to Roundup showed a poorer sperm quality, measured as reduced plasmatic membrane integrity, mitochondrial functionality, DNA integrity, motility, motility period and concentration of spermatic cells, than those kept under control condition (no Roundup addition to the water). Most of the spermatic parameters analyzed showed strong association to each other, which may help to understand the mechanisms underlying the observed reduction in sperm quality. Exposure to Roundup did not alter the biochemical parameters analyzed, though differences between genders were observed and deserve further investigations. Findings from the present study suggest that exposure to environmentally relevant concentrations of Roundup may negatively affect at long-term the reproduction of P. vivipara, with consequent changes in fish populations inhabiting environments contaminated with the herbicide.