The influence of gamete co-incubation length on the in vitro fertility and sex ratio of bovine bulls with different penetration speed

The objectives of this work were to evaluate whether the sperm penetration speed is correlated to the in vitro fertility and whether adapting the gamete co-incubation length to the kinetics of the bull improves in vitro fertility and affects the sex ratio. In vitro matured oocytes were co-incubated with spermatozoa from four different bulls (A-D). At various post-insemination (p.i.) times (4, 8, 12, 16 and 20 h), samples of oocytes were fixed and stained with DAPI for nuclei examination, while the remaining ones were transferred into culture to evaluate embryo development. The blastocysts produced were sexed by PCR. Two bulls (A and B) had faster kinetics than the others (C and D), as shown by the higher penetration rates recorded at 4 h p.i. (43%, 30%, 11% and 6%, respectively for bulls A, B, C and D; p<0.01). The differences in the kinetics among bulls did not reflect their in vitro fertility. The incidence of polyspermy was higher for faster penetrating bulls (36%, 24%, 16% and 4%, respectively for bulls A, B, C and D; p<0.01) and at longer co-incubation times (0%, 16%, 19%, 30% and 34%, respectively at 4, 8, 12, 16 and 20 h p.i.; p<0.01). The fertilizing ability of individual bulls may be improved by adapting the co-incubation length to their penetration speed. A sperm-oocyte co-incubation length of 8 h ensured the greatest blastocyst yields for the two faster penetrating bulls. On the contrary, 16 h co-incubation was required to increase (p<0.01) cleavage rate of the two slower bulls. Bulls with a faster kinetics did not alter the embryo sex ratio towards males. The female/male (F/M) ratios recorded were 2.1, 1.4, 1.2, 1.3 and 1.6, respectively at 4, 8, 12, 16 and 20 h p.i.     

Closed cycle subirrigation with low concentration nutrient solution can be used for soilless tomato production in saline conditions

Closed cycle soilless techniques can be adopted to minimize water and fertilizer losses in greenhouse cultivation. There is a general lack of information regarding the soilless cultivation of vegetables with closed cycle subirrigation techniques, specifically when using saline water. In this study, a trough bench subirrigation system (SUB), with two fertilizer concentrations (“100%”, containing 9.8 mol m⁻³ N-NO₃, 1.6 mol m⁻³ P-H₂PO₄, 8.7 mol m⁻³ K⁺, 2.8 mol m⁻³ Ca⁺, 1.8 mol m⁻³ Mg⁺, 4 mol m⁻³ S-SO₄, and “70%”, containing 70% of the macronutrient concentration) in the nutrient solution (NS), was compared with open cycle drip-irrigation (DRIP with “100%” NS). For all the three treatments, NS was prepared using rain water (0.05 dS m⁻¹) and adding NaCl (1 g L⁻¹), in order to simulate moderate saline irrigation water. The effect of the treatments on tomato (Solanum lycopersicum L.) plant growth, yield, fruit quality, water use efficiency (WUE) and fertilizer consumption was evaluated. Substrate and recirculating NS composition were also studied. Subirrigation, regardless of NS concentration, reduced plant height (by 30 cm), leaf area (by 1411 cm²), total fresh and dry weight (by 429 and 48.5 g plant⁻¹, respectively) but not dry matter percentage of the whole plant, with respect to DRIP. Yield was reduced when plants were subirrigated with the higher concentrated NS, but no differences with open cycle DRIP were recorded when the lower NS concentration was used in SUB. Fruit quality was not affected by irrigation system or NS concentration. The higher WUE was obtained with subirrigation. NaCl accumulated similarly over the crop cycle in recirculating NS of both SUB treatments and in growing substrates of all the three treatments. Higher salt concentration was found in subirrigated substrates, in particular in the upper part of the substrate profile. Fertilizers accumulated in the subirrigated substrates when the higher NS concentration was used, but not when the NS concentration was reduced by 30%. The results of this study indicate that tomato can be grown successfully in a closed cycle subirrigation system, using saline water, by reducing the fertilizer NS concentration normally used with traditional open cycle systems.     

DIAGNOSTIC SENSITIVITY OF ULTRASOUND,RADIOGRAPHY AND COMPUTED TOMOGRAPHY FOR GENDER DETERMINATION IN FOUR SPECIES OF LIZARDS

Gender determination is frequently requested by reptile breeders, especially for species with poor or absent sexual dimorphism. The aims of the current study were to describe techniques and diagnostic sensitivities of ultrasound, radiography, and computed tomography for gender determination (identification of hemipenes) in four species of lizards. Nineteen lizards of known sex, belonging to four different species (Pogona vitticeps, Uromastyx aegyptia, Tiliqua scincoides, Gerrhosaurus major) were prospectively enrolled. With informed owner consent, ultrasound, noncontrast CT, contrast radiography, and contrast CT (with contrast medium administered into the cloaca) were performed in conscious animals. Imaging studies were reviewed by three different operators, each unaware of the gender of the animals and of the results of the other techniques. The lizard was classified as a male when hemipenes were identified. Nineteen lizards were included in the study, 10 females and nine males. The hemipenes were seen on ultrasound in only two male lizards, and appeared as oval hypoechoic structures. Radiographically, hemipenes filled with contrast medium appeared as spindle‐shaped opacities. Noncontrast CT identified hemipenes in only two lizards, and these appeared as spindle‐shaped kinked structures with hyperattenuating content consistent with smegma. Hemipenes were correctly identified in all nine males using contrast CT (accuracy of 100%). Accuracy of contrast radiography was excellent (94.7%). Accuracy of ultrasound and of noncontrast CT was poor (64.3% and 63.1%, respectively). Findings from the current study supported the use of contrast CT or contrast radiography for gender determination in lizards.     

Persistence of benfluralin in soil and its uptake by carrots

Analytical methods are described for the determination of residues of benfluralin in soil and carrots. All plots treated with 1.3 or 1.9 kg benfluralin/ha produced average carrot yields significantly higher than untreated plots. Degradation of benfluralin in soil was very slow during the winter. As the temperature rose, the rate of benfluralin decline was accelerated; 324 days after incorporation, benfluralin residues were reduced to about 11 and 14% of the applied dose. The average total residues of benfluralin found in whole carrots were 210 ± 18 and 240 ± 47 ng/g following doses of 1.3 and 1.9 kg a.i./ha respectively. The peel of carrots contained 92% of the total residue, and there seemed to be little translocation of the compound through the plant tissues.     

Gas-liquid chromatographic method for perfluidone residues in rice grains and husks

A gas-liquid chromatographic method using electron-capture detection is described for the determination of perfluidone residues in rice grains and husks. The procedure consists of extraction, purification on an anion-exchange resin, methylation and further clean-up (if necessary) by adsorption on “Florisil”. The recoveries from fortified rice grains and husks were respectively 73.5–98% and 77–104% in the range 0.01–0.50 mg/kg. Sensitivity permits detection of perfluidone at 0.005 mg/kg level.     

Luteoforol,a flavan 4-ol,is induced in pome fruits by prohexadione-calciumand shows phytoalexin-like properties against <Emphasis Type="Italic">Erwinia amylovora</Emphasis>and other plant pathogens

Treatments with prohexadione-calcium led to lowered incidence of fire blight, scab and other diseases in pome fruit trees and other crop plants. In addition to acting as a growth regulator, prohexadione-calcium interferes with flavonoid metabolism and induces the accumulation of the 3-deoxycatechin luteoliflavan in shoots of pome fruit trees. Luteoliflavan does not possess any remarkable antimicrobial activity. Therefore luteoforol, its unstable and highly reactive precursor, has been tested in vitro for its bactericidal and fungicidal activities. Luteoforol was found to be highly active against different strains of Erwinia amylovora, the causal agent of fire blight, and all other bacterial and fungal organisms tested. Phytotoxic effects were also observed in pear plantlets. The results obtained indicate that prohexadione-calcium induces luteoforol as an active principle with non-specific biocidal properties. It is proposed that luteoforol is released upon pathogen attack from its cellular compartment and inhibits further disease development by destroying pathogen cells as well as by inducing a hypersensitive-like reaction in the host plant tissue. This mechanism would be closely analogous to the one known for structurally related phytoalexins in sorghum.     

Influence of some 1,3,5-triazine herbicides on peroxidase activity in corn seedlings

The effect of some 1,3,5-triazine herbicides, on the rate of oxidative decarboxylation of tryptophan by peroxidase from corn seedlings, varied with the concentration. Atrazine was stimulatory in vivo at 10-⁶M but not at 10⁸ M or lower, whereas terbutryne was inhibitory at 10-⁶M but not at 10-⁸M or below. Prometryne inhibited enzyme activity at 10-⁶M and stimulated it at 10-¹⁰M and at 10¹²M, while simazine caused stimulation at 10-⁶M and inhibition at 10^?10M and 10^?12M. Atraton reduced enzyme activity at 10^?6M and 10^?8M but had no effect at 10^?10M and 10^?12M. When peroxidase activity was assayed in acetone-dried powders from seeds, harvested from fields that had received typical atrazine applications, the quantities of tryptophan consumed were significantly less than the quantity determined in the control powder. From the results obtained, by incorporating dealkylated and 2-hydroxy metabolites of atrazine at 10^?6M into the culture solutions, it seems possible that the inhibition observed may have been due to the presence of 4-amino-6-ethylamino-1, 3, 5-triazin-2-ol and/or 4, 6-diamino-1, 3, 5-triazin-2-ol residues in seeds from treated plots, even if, in previously reported experiments, analyses had not detected their presence.