Effect of Several Antioxidants on Thawed Ram Spermatozoa Submitted to 37°C up to Four Hours

Thawed ram spermatozoa were incubated at 37°C in the presence of dehydroascorbic acid (DHA), TEMPOL (TPL), N‐acetyl‐cysteine (NAC) and rutin (RUT), at 0.1 and 1 mm , in order to test their effects on sperm physiology. Cryopreserved spermatozoa from four rams were thawed, pooled, washed and incubated in TALP‐Hepes with 1 mm or 0.1 mm of each antioxidant, performing a replicate with induced oxidative stress (Fe²⁺/ascorbate). Motility (CASA), viability and mitochondrial membrane potential (flow cytometry) were analysed at 2 and 4 h. Lipoperoxidation (MDA production), intracellular reactive oxygen species (ROS) and DNA status (TUNEL) were analysed at 4 h. Antioxidants, except DHA 0.1 mm , decreased motility and kinematic parameters, but had little effect on viability or mitochondrial activity. Except 1 mm DHA, the antioxidants reduced ROS at 4 h. Moreover, NAC 1 mm , rutin and TEMPOL reduced ROS and DNA damage in the presence of oxidative stress. N‐acetyl‐cysteine, rutin 1 mm and TEMPOL reduced lipoperoxidation in the presence of oxidative stress. However, DHA did not affect lipoperoxidation. At 1 mm , DHA increased DNA damage in the absence of oxidative stress. Dehydroascorbic acid effects could arise from spermatozoa having a low capacity for reducing it to ascorbic acid, and it may be tested in the presence of other antioxidants or reducing power. Future research should focus in testing whether the inhibition of motility observed for NAC, rutin and TEMPOL is reversible. These antioxidants might be useful at lower temperatures (refrigerated storage or cryopreservation) when their protective effects could be advantageous.     

In vitro screening of plant lectins and tropical plant extracts for anthelmintic properties

Lectins are plant secondary metabolites (PSM) found in many forages and which may confer anthelmintic properties to gastrointestinal parasites through disrupting the development of parasitic larvae throughout its life cycle. In experiment 1, the ability of the plant lectins jacalin (JAC), concanavalin A (Con A), phytohemagglutinin E2L2 (PHA-E2L2), phytohemagglutinin L4 (PHA-L4), phytohemagglutinin E3L (PHA-E3L), kidney bean albumin (KBA), Robinia pseudoacacia agglutinin (RPA), Maackia amurensis lectin (MAA), Maclura pomifera agglutinin (MAA), Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA) and Galanthus nivalis agglutinin (GNA) to disrupt the feeding of the first stage larvae (L(1)) of the sheep gastro-intestinal nematodes (GIN) Teladorsagia circumcincta, Haemonchus contortus and Trichostrongylus colubriformis was investigated using a larval feeding inhibition test (LFIT). Only PHA-E3L, WGA and Con A had a potent effect on disrupting larval feeding of all of the three species of GIN investigated. The lectin concentration required to inhibit feeding in 50% of L(1) (IC50) was 7.3±1.2, 8.3±1.4 and 4.3±1.7 μg/ml for PHA-E3L; 59.1±32.4, 58.7±11.9 and 8.1±7.0 μg/ml for Con A and 78.9±11.2, 69.4±8.1 and 28.0±14.1 μg/ml for WGA for T. circumcincta, H. contortus and T. colubriformis larvae, respectively (P=0.006). The addition of the lectin inhibitors fetuin, glucose/mannose or N-acetylglucosamine for PHA-E3L, Con A and WGA, respectively, caused an increase in the proportion of larvae that had fed at all concentrations for PHA-E3L only. In experiment 2, the effect of extracts from the tropical plants Azadiractha indica, Trichanthera gigantea, Morus alba, Gliricidia sepium and Leucaena leucocephala on the feeding behaviour of H. contortus L(1,) was examined. A. indica, T. gigantea and M. alba failed to inhibit 50% of larvae from feeding at concentrations up to 10mg plant extract per ml. In contrast, both G. sepium and L. leucocephala demonstrated a dose-dependent effect on larval feeding with respective IC50 estimates (mean±s.e.) of 0.015 mg/ml ±0.001 and 3.465 mg/ml ±0.144, effects which were partly reversed by the inclusion of either the tannin inhibitor polyethylene glycol or the lectin inhibitor Fetuin. These studies demonstrate that plant lectins can have an inhibitory effect on the feeding behaviour of first stage larvae of ovine GIN in vitro. Moreover they also provide novel evidence that lectins may contribute to the anthelmintic properties of some tropical forage plant extracts, such as G. sepium and L. leucocephala.     

Position of the head is not associated with changes in horse vision

It has become accepted that the horse cannot see directly in front when the nose is lowered and must therefore rely on the rider. We tested the hypothesis that this conclusion would be correct only if the horse did not adjust the eyeball horizontal axis to changes of the head position. The results of the present study suggest that it is unlikely that horses have limited vision in relation to their head position when driven by the rider, and that the horse maintains the optimal horizontal eyeball position regardless of head position relative to the ground.     

Feather content of porphyrins in Eurasian eagle owl (Bubo bubo) fledglings depends on body condition and breeding site quality

Porphyrins are pigments produced in most animal cells during the synthesis of heme, but their importance for external coloration is unclear. Owls (Order Strigiformes) are among the few animals that accumulate porphyrins in the integument, where it could serve as a means of signaling. Here we hypothesized that the porphyrin content of feathers may depend on body condition and breeding site quality in Eurasian eagle owl (Bubo bubo) fledglings and, thus, constitute amplifiers of the quality of the area where they are born. Using high‐performance liquid chromatography, we found 2 porphyrins (protoporphyrin IX and coproporphyrin III) in the body feathers of 19 eagle owl fledglings from 7 breeding territories. Coproporphyrin III, but not protoporphyrin IX feather concentration, was positively associated with the body mass of fledglings and with the quality of the breeding sites where they were reared with respect to food quality and availability. As coproporphyrin III is produced under oxidative stress, we suggest that good breeding sites may lead to fledglings in good condition. This, in turn, may make fledglings induce a certain level of free radical and coproporphyrin III production to signal to conspecifics their site‐mediated capacity to cope with oxidative stress. This is the first time that porphyrin content in the integument has been found to be related to individual quality, opening a new scenario for studying evolution of animal coloration.     

Pathogenic profile and cytotoxic activity of Aeromonas spp. isolated from Pectinatella magnifica and surrounding water in the South Bohemian aquaculture region

Pectinatella magnifica is an invasive freshwater bryozoan that has expanded in many localities worldwide, including fishing areas. It contains microbial communities, predominantly consisting of Aeromonas bacteria that are frequently associated with fish infections. The objective of this study was to investigate the potential pathogenicity of Aeromonas spp. associated with P. magnifica and evaluate the health risks for fish. Aeromonas strains were isolated from P. magnifica (101 strains) and from surrounding water (29 strains) in the South Bohemian region and investigated for the presence of 14 virulence-associated genes using PCR. We demonstrated high prevalence of phospholipase GCAT, polar flagellin, enolase, DNAse, aerolysin/cytotoxic enterotoxin, serine protease and heat-stable cytotonic enterotoxin-coding genes. Further, all twelve isolates that were analysed for cytotoxicity against intestinal epithelial cells were found to be cytotoxic. Six of the isolates were also tested as co-cultures composed of pairs. Enhanced cytotoxicity was observed when the pair was composed of strains from different species. In conclusion, P. magnifica is colonized by Aeromonas strains that have a relatively high prevalence of virulence-associated genes and the ability to provoke disease. Results also suggest a possibly increased risk arising from mixed infections.     

Effect of Streptomyces sp. RL8 on the survival of Artemia franciscana nauplii and resistance to Vibrio parahaemolyticus

Fisheries Science - Artemia franciscana nauplii were enriched with Streptomyces sp. RL8 in order to assess its effect on the survival rate of Artemia and its protective effects against Vibrio...     

Activity,molecular mass and hydrolysis on baker's yeast protein of extracellular proteases from the putative probiotic bacteria Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N

The bacteria Microbacterium sp. 8L and Exiguobacterium mexicanum 8N are known to improve the culture of Artemia franciscana using baker's yeast as food. Using spectrophotometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE), substrate‐SDS‐PAGE and pH‐stat in vitro‐digestibility assays, the activity, molecular mass and hydrolysis on baker's yeast protein of proteases from extracellular polymeric substances (EPS) of the strains 8L and 8N along with the pathogenic strains Microbacterium sp. 8R and Vibrio parahaemolyticus 588 CECT (Vp) were studied. The EPSs of 8L and 8R showed one activity band, on which the serine inhibitor phenylmethylsulphonyl fluoride (PMSF) had no effect. The EPSs of 8N showed four bands; two were unaffected by PMSF, whereas one was affected, and the other was partially affected. The EPSs of Vp showed two bands, one partially inhibited by PMSF. No inhibitory effects from 1‐chloro‐3‐tosylamido‐7‐amino‐2‐heptanone (trypsin inhibitor) were observed in the protease bands of the studied bacteria. The EPSs of 8L and 8N showed a similar degree of hydrolysis (pH‐stat). The EPSs of 8L had the lowest Dice index of similarity of yeast protein profiles at 1 h of reaction. We conclude that the strain 8L could benefit A. franciscana by providing bacterial proteases for digestion of baker's yeast.     

Commercial‐scale Validation of Temperature‐step Rearing on Growth Physiology in Turbot,Scophthalmus maximus

The aim of this study was to investigate the possible benefit of “temperature‐steps” (T‐steps) rearing for juvenile turbot (initial weight 15.1 g) under realistic production scale and to determine whether initial growth advantage is maintained throughout the rearing period to market size. One group (called T‐step 22‐19‐16) of juvenile turbot was reared at three different temperatures, that is, 22 C (from 17 to 60 g) followed by 19 C (from 60 to 100 g) and 16 C (>100 g); another group (called T‐step 19‐16) at two temperatures, that is, 19 C (from 17 to 100 g) and lowered to 16 C (>100 g); and the third group (called C16) at one constant temperature, that is, 16 C. Relative growth was significantly higher in the two T‐step groups, with the T‐step 19‐16 showing the highest overall growth. Feed conversion efficiency was highest in the 19‐16 group. Only minor effects of the experimental rearing on blood physiology were found, with one notable exception of inverse relationship between plasma glucose and growth. Overall, these findings indicate that a short interval of rearing fish at high temperature during the early juvenile phase may have a long‐term effect on biomass increment in turbot. This is an important finding for the turbot industry.     

The Use of Thermography Images in the Description of the Humidification Bulb in Soilless Culture

Thermography is a tool used in many scientific disciplines, including agriculture. Determining the behavior of a nutritive solution in soilless culture containers using substrates is very important in order to establish the position of drippers, drainage holes, the use of humidification agents, and fertigation management in culture units. This paper describes the application of thermography to determine the humidification bulb in horticulture crops to rapidly diagnose the best way to advance fertigation in different horticultural substrates in soilless culture. Thermography was seen to be an excellent tool for evaluating the movement and distribution of the nutritive solution within the soilless culture container with different substrates.     

Effect of the Extrusion on Functional Properties and Mineral Dialyzability from <Emphasis Type="Italic">Phaseolus Vulgaris</Emphasis> Bean Flour

The effects of extrusion conditions on cooking degree, flour dispersion viscosity and mineral potential availability of extruded bean flour were studied. Phaseolus vulgaris beans of the agronomic cultivar “Flor de mayo” were ground and dehulled to obtain grits and then extruded at different temperatures (140, 160 and 180 °C) and moisture contents (17, 20 and 23%), according to a bifactorial experimental design. Degree of cooking was estimated by water solubility (WS) and specific mechanical energy (SME). The effect of variables on WS and SME were analysed by surface response methodology. Flour dispersion viscosity and mineral availability (estimated by in vitro dialyzability), were also evaluated on selected samples. Results showed that, within the ranges of the variables used for this study, only the effect of temperature was significant on the degree of cooking. No direct correlation was observed between water solubility and SME, although a maximum value of WS corresponded to a range of SME values of 400–500 J/g was observed. Dispersion viscosity decreases as WS increases, so if high calorie density is desired, for instance in order to produce a cream soup formula, bean grits should be extruded at high temperature and as low moisture as possible, in our case 180 °C and 17% moisture. On the other hand, the effects of extrusion variables on iron and zinc dialyzability were not much affected.