Initial cooling time before freezing affects post‐thaw quality and reproductive performance of rabbit semen

The aim of this study was to investigate the effect of initial cooling time at 5°C during semen cryopreservation on post‐thaw quality and reproductive performance of rabbit semen. Pooled semen samples (n = 6) were divided into two subsamples and cooled at 5°C for 45 or 90 min. After cooling, the semen samples were diluted to a ratio of 1:1 (v:v) with a freezing extender composed of Tris‐citrate‐glucose (TCG) containing 16% of dimethylsulfoxide and 0.1 mol/L sucrose. The semen was subsequently loaded in 0.25 ml straws, equilibrated at 5°C and frozen in liquid nitrogen vapor. After thawing, sperm motility, viability, osmotic resistance, acrosome and DNA integrity were assessed. Our results indicate that the longer cooling time, that is, 90 min before cryopreservation significantly improves sperm post‐thaw viability, motility and fertility. In fact, reproductive performances obtained with semen frozen after a 90 min cooling time were similar to those produced by fresh semen insemination. Hence, the present research provides an effective freezing protocol for rabbit semen that will allow for the creation of a sperm cryobank for the conservation of Italian rabbit genetic resources, as well as the use of frozen semen doses in commercial farms.     

Is crop N demand more closely related to dry matter accumulation or leaf area expansion during vegetative growth?

The critical crop nitrogen uptake is defined as the minimum nitrogen uptake necessary to achieve maximum biomass accumulation (W). Across a range of crops, the critical N uptake is related to W by a power function with a coefficient less than unity that suggests crop N uptake is co-regulated by both soil N supply and biomass accumulation. However, crop N demand is also often linearly related to the expansion of the leaf area index (LAI) during the vegetative growth period. This suggests that crop N demand could be also linked with LAI extension. In this paper, we develop theory to combine these two concepts within a common framework. The aim of this paper is to determine whether generic relationships between N uptake, biomass accumulation, and LAI expansion could be identified that would be robust across both species and environment types. To that end, we used the framework to analyze data on a range of species, including C₃ and C₄ ones and mono- and di-cotyledonous crops. All crops were grown in either temperate or tropical and subtropical environments without limitations on N supply. The relationship between N uptake and biomass was more robust, across environment types, than the relationship of LAI with biomass. In general, C₃ species had a higher N uptake per unit biomass than C₄ species, whereas dicotyledonous species tended to have higher LAI per unit biomass than monocotyledonous ones. Species differences in N uptake per unit biomass were partly associated with differences in LAI and N-partitioning. Consequently the critical leaf-N uptake per unit LAI (specific leaf nitrogen, SLN) was relatively constant across species at 1.8–2.0 g m⁻², a value that was close to published data on the critical SLN of new leaves at the top of the canopy. Our results indicate that critical N uptake curves as a function of biomass accumulation may provide a robust platform for simulating N uptake of a species. However, if crop simulation models are to capture the genotypic and environmental control of crop N dynamics in a physiologically functional manner, plant growth has to be considered as the sum of a metabolic (e.g. leaves) and a structural (e.g. stems) compartment, each with its own demand for metabolic and structural N.     

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.     

Antinociceptive effect and acute toxicity of the Hyptis suaveolens leaves aqueous extract on mice

The aqueous extract of Hyptis suaveolens leaves was studied for their antinociceptive property in chemical and thermal models of nociception in mice. Oral administration of the aqueous extract (100, 200, and 400 mg/kg) dose-dependently reduced the number of writhings induced by acetic acid, decreased the licking activity of the early phase in formalin test and increased the reaction time in hot-plate test. The antinociceptive effect was significantly antagonized by naloxone (3 mg/kg; i.p.). Preliminary acute toxicity study showed that no animal death with doses up to 5 g/kg (p.o.).     

Genotoxic activity of dichlorvos,trichlorfon and dichloroacetaldehyde

Two structurally related organophosphorous insecticides, dichlorvos and trichlorfon, and their main metabolite dichloroacetaldehyde, were assayed for their ability to induce DNA repair, detected as unscheduled DNA synthesis in human epithelial-like cell (EUE) cultures in vitro. A dose-response relationship was found for treatment with the two pesticides, but negative results were obtained with dichloroacetaldehyde. Tests for induction of gene mutation, as ouabain resistance in Chinese hamster cells (V79), failed to show any mutagenic activity by these compounds.     

Synthesis and juvenile hormone activities of some new ether derivatives of hydroquinone

A new class of compounds, derived from hydroquinones, has been synthesised and bioassayed for juvenile hormone activity on yellow mealworm (Tenebrio molitor), confused flour beetle (Tribolium confusum), and mediterranean flour moth (Anagasta kuehniella). Structure-biological activity relationships of the most significant products are tabulated and discussed. Two of the new compounds, 1, 4-bis(5-chloropent-4-ynyloxy) benzene (I) and 1-(5-chloropent-4-ynyloxy)-4-phenoxybenzene (II), showed outstanding activity in morphogenetic assay with T. molitor, and good field persistence. Product II is of particular interest for the control of various Homopteran species, mosquitoes, stored product pests and fire ants.     

New chlorinated juvenile hormone analogues and their biological activity

In the search for new juvenile hormone analogues (JHAs), a general approach was chosen based on the substitution of the isoprene unit, that carries the epoxy function in the natural hormone JHIII, by 3,3,3-trichloropropyl, 2,2-dichlorocyclopropyl- methyl, 3,3-dichloroallyl, or 3-chloroprop-2-ynyl groups. Accordingly, several terpenoid aromatic JHAs were prepared with, or without, oxygen atoms in the aliphatic chain. Structure-biological activity relationships of the most active compounds toward the yellow mealworm (Tenebrio molitor) and the large cabbage white butterfly (Pieris brassicae) are given. With a view to preparing synthetically accessible analogues, some alkyl phenyl ethers were also made, which were not branched in the aliphatic chain and were related terminally to the most active compounds. They were only moderately active against insects when tested for morphogenetic activity, but exhibited a good field persistence. The occurrence of a moderate acaricidal activity among these analogues prompted the examination of several 1,4-disubstituted-phenyl derivatives. The ovicidal effectiveness of these products against a susceptible strain of the two-spotted spider mite, Tetranychus urticae, was ten or 100 times higher than that of the well known and highly potent JHAs.     

Molecular characterization of Fusarium oxysporum f.sp. cichorii pathogenic on chicory (Cichorium intybus)

Fusarium oxysporum is a ubiquitous soilborne ascomycete responsible for vascular wilt in many plant species worldwide. This species comprises more than 120 putative host-specific formae speciales capable of causing marked economic losses. In summer 2009, wilt symptoms, including chlorosis and poor development of the root system, were observed on cultivars of chicory (Cichorium intybus) in northern Italy. The causal agent isolated from symptomatic tissues in this case was identified as F. oxysporum on the basis of both morphological features and molecular analyses. In this work, we attempted to characterize the isolates of F. oxysporum from C. intybus by both biological and molecular approaches. Pathogenicity trials performed on five species of the Asteraceae family with isolates of F. oxysporum from C. intybus indicated that the pathogen has a unique host range, infecting chicory only. Neither lettuce nor endive, lawn daisy or Paris daisy developed the disease. Five cultivars within C. intybus species were tested, and the cv. ??Clio?? was the most susceptible. Phylogenetic analyses relative to the ribosomal intergenic spacer (IGS) and translation elongation factor 1-alpha (EF1-??) assigned isolates pathogenic to chicory to a single cluster, distinct from other pathogenic F. oxysporum. In light of these findings, we propose to designate this organism as Fusarium oxysporum f.sp. cichorii.     

Conventional and real-time PCR for the identification of Fusarium fujikuroi and Fusarium proliferatum from diseased rice tissues and seeds

Fusarium fujikuroi is a species of the Gibberella fujikuroi species complex (GFSC) and the causal agent of bakanae disease on rice. Even if F. fujikuroi is the most abundant Fusarium species found on rice, other species can also be isolated from rice, such as F. proliferatum. Multiple alignment of translation elongation factor (TEF) gene sequences of different Fusarium spp., showed a deletion of six nucleotides in F. fujikuroi sequence and a two nucleotide polymorphism in the same region of F. proliferatum sequence. These elements of variability were used to develop a conventional and Real-Time PCR assay for diagnosis. The species specific primer pairs (Fuji1F/TEF1R and Proli1F/TEF1R) gave a product of 179 and 188?bp for F. fujikuroi and F. proliferatum respectively. Primer specificity was confirmed by analyzing the DNA of the most representative species of the GFSC and 298 strains of Fusarium spp. isolated from rice plants and seeds in Italy. The specific primers were also successfully used to detect fungal presence directly from infected rice tissues and seeds, providing a rapid tool for the early detection of pathogen contamination.