Quality and Fertilizing Ability In Vivo of Sex‐Sorted Stallion Spermatozoa

Little information is available on the quality of stallion spermatozoa after sex sorting. The objectives of the present study were to assess the quality of sex‐sorted stallion spermatozoa and determine its fertilizing ability after hysteroscopic low dose insemination. Ejaculates from four stallions were collected and sorted by a MoFlo SX^® flow cytometer/sperm sorter. Before and after sorting, spermatozoa were evaluated for motility by Computer Assisted Sperm Analysis, viability (SYBR 14‐propidium iodide), mitochondrial function (JC‐1) and acrosomal status (fluorescein isothiocyanate Pisum sativum agglutinin conjugated). A fertility trial was carried out on four mares (seven oestrous cycles) by hysteroscopic insemination, depositing 5 × 10⁶ X‐bearing spermatozoa. Sex sorting resulted in a significant decrease (p < 0.001) in all motility characteristics. Sperm viability and percentage of spermatozoa with functional mitochondria were not affected by the sorting process, while the percentage of reacted spermatozoa was higher (p < 0.01) for non‐sorted than sorted spermatozoa. Pregnancy rate was 28.6% (2/7) after low dose hysteroscopic insemination. Only one pregnancy was carried to term with the birth of a healthy filly. In conclusion, despite the reduction in sperm motility, sex sorting did not impair stallion sperm viability and mitochondrial activity immediately post‐thaw; moreover, the sexed spermatozoa retained the ability to fertilize in vivo.     

Arbuscular mycorrhizal inoculation of micropropagated persimmon plantlets

Summary

Growth, development and nutrient status of micropropagated persimmon (Diospyros kaki) ‘Rojo Brillante’ in response to inoculation with two species of arbuscular mycorrhizal (AM) fungi were studied under nursery conditions. The species of AM fungi were Glomus intraradices and G. mosseae. Shoot growth depression and low root colonization percentage were observed to G. mosseae inoculation. Shoot and root growth enhancements were observed for plants colonised by G. intraradices. Inoculated plantlets with G. intraradices and high level of controlled-release fertilizer signi®cantly decreased shoot height in relation to treatment with low fertilization level. Furthermore, G. intraradices had unique effects on the mineral status of the persimmon plantlets. N and Ca were significantly increased and decreased, respectively, within the shoot tissue of plants colonised by G. intraradices. Early inoculation with G. intraradices appears to favour growth of micropropagated persimmon plantlets.     

Epicatechin-3-O-(3″-O-methyl)-gallate content in various tea cultivars (Camellia sinensis L.) and its in vitro inhibitory effect on histamine release

It has been reported that epigallocatechin-3-O-(3″-O-methyl)-gallate (EGCG3″Me) and the EGCG3″Me-rich green tea ( Camellia sinensis L.) cultivar 'Benifuuki' exhibit antiallergic effects. The objective of this study was to investigate the effect of various tea leaf catechins on histamine release from murine bone marrow mast cells (BMMC). At a dose of 50 μg/mL, the rank order of histamine release inhibition was observed to be epicatechin-3-O-(3″-O-methyl)-gallate (ECG3″Me) > gallocatechin-3-O-(3″-O-methyl)-gallate (GCG3″Me) > EGCG3″Me > gallocatechin-gallate (GCG) > catechin-gallate (CG) > EGCG > epicatechin-gallate (ECG) > epigallocatechin (EGC) > gallocatechin (GC). Of the various tea cultivars analyzed by HPLC, the greatest content of ECG3″Me was found in the third crop of 'Benifuuki' (1.05% dry weight). Moreover, ECG3″Me content was positively correlated with EGCG3″Me content in 'Benifuuki' tea leaves. In an assay of mixtures of ECG3″Me and EGCG3″Me, inhibitory activity (50 μg/mL in total) was increased as the content of ECG3″Me increased. This suggests that ECG3″Me might link to the antiallergic effect of 'Benifuuki' tea, as has been reported for EGCG3″Me.     

The quality of cryopreserved sperm collected from feline caudal epididymides stored at room temperature

On the assumption that animals of wild feline species died in the field, caudal epididymal sperm were cryopreserved following storage of the feline epididymides at 20°C for 0-24 hr, and their qualities were observed. Compared to the qualities at 0 hr, no significant differences were noted following 12 hr of storage at 20°C. On comparison of the qualities between caudal sperm cryopreserved after 24 hr storage at 4°C and after 12 hr at 20°C followed by 12 hr storage at 4°C, no significant differences were noted. These findings suggest that the cryopreserved sperm collected from epididymides of dead animals might be useful for artificial insemination if cryopreservation was performed within 12 hr exposure to ambient temperature.     

Do carbon farming practices build bioavailable nitrogen pools?

Agricultural soils contain large amounts of nitrogen (N), but only a small fraction is readily available to plants. Despite several methods developed to estimate the bioavailability of N, there is no consensus on which extraction methods to use, and which N pools are critically important. In this study, we measured six soil N pools from 20 farms, which were part of a multi-year soil carbon sequestration on-farm experiment (Carbon action, 2019–2023). The aim was to quantify the N pools and to evaluate if farming practices that aim to build soil carbon pools, also build bioavailable N pools. We also aimed to test if the smaller and rapidly changing N pools could serve as an indicator for the slower change in soil organic matter. The measured N pools decreased in size, when moving from total N (7700 ± 1500 kg/ha) to slowly cycling (Illinois Soil Nitrogen Test ISNT-N: 1063 ± 220 kg/ha, autoclave citrate-extracted ACE protein N: 633 ± 440 kg/ha), water-soluble organic N (50 ± 17 kg/ha), potentially mineralizable N (33 ± 13 kg/ha) and finally readily plant available inorganic pools (nitrate and ammonium, total: 14 ± 8 kg/ha). In total, the measured pools covered only 18%–44% of total N, indicating a large unidentified N pool, which is either tightly bound to soil mineral fraction and not easily extractable or is bound to undecomposed plant residues and not hydrolysed by the methods. Of the large N pools (ISNT-N, ACE protein and unidentified residual N), clay, carbon (C) and C:Clay ratios explained most of the variability (R² = .90–.93), leaving a minor part of the variation to the management effect. A pairwise comparison of carbon farming and control plots concluded that farming practices had a small (3%–5%) but statistically significant (p < .05) effect on soil total N and ISNT-N pools, and a moderate and significant effect (18%, p < .01) on potentially mineralizable N. The large variation in protein N, water-soluble organic N and inorganic N reduced statistical significance, although individual C sequestration practices had large effects (−30% to +50%). In conclusion, carbon sequestration practices can build both slowly cycling N pools (ISNT) and increase the mineralisation rate of these pools to release plant available forms, resulting in an additional benefit to agriculture through reduced fertilizer application needs.     

Effects of Broomrape Parasitism on Sunflower Plants: Growth,Development, and Mineral Nutrition

Sunflower broomrape (Orobanche cumana Wallr.) is a parasitic plant that infects sunflower (Helianthus annuus L.) plants. In this work, sunflower plants were grown under greenhouse conditions in pots with the substrate infested or non-infested with broomrape seeds. At different numbers of days after sowing, plant height, internode lengths, number of leaves, head diameter, mineral composition of leaves, and potassium (K) concentration in stem were measured. The negative effects of broomrape parasitism were assessed from 57 d after sowing, when broomrape started to emerge. Parasitized plants exhibited lower shoot dry weight, height, and head diameter than control plants. The reduction in internode lengths was associated with a decrease in the gradient of K concentration from basal to apical stem. The mineral composition of leaves was also affected in parasitized plants. The concentrations of calcium (Ca), magnesium (Mg), manganese (Mn), and zinc (Zn) in leaves of parasitized plants were lower than those of the control plants, while there were few differences for K, phosphorus (P), iron (Fe), and copper (Cu). The effects of parasitism are discussed in relation to their competition for resources and to perturbations of the host physiology such as hormonal and water balance.     

Application of a plant bioassay for the evaluation of ecotoxicological risks of heavy metals in sediments affected by mining activities

Purpose

The objective of this work was to evaluate the effectiveness of a plant bioassay (Phytotoxkit®) for screening ecotoxicological risks in sediments affected by mining activities.

Materials and methods

A total of 42 sediment samples affected by mining activities were studied, including 39 sediment samples from the Sierra Minera, Spain, an area affected by old extraction procedures, and three sediments from an area affected by opencast mining. These three samples were then mixed with limestone filler at 10, 20 and 30 %, providing nine stabilised samples. The total and soluble metal(loid) content (As, Cd, Cu, Fe, Pb and Zn) was determined in all samples, and the Phytotoxkit® bioassay was applied to determine the ecotoxicological effect of this procedure.

Results and discussion

The stabilised material had a neutral pH and low soluble metal(loid) concentration, similar to that of samples in which a natural attenuation process had taken place because of mixing with surrounding carbonate-rich materials. An ecotoxicological survey identified the low toxicity levels of the stabilised samples.

Conclusions

The applied bioassay is a good tool for screening metal(loid) contamination in areas affected by mining activities, since it provides information on both natural and simulated attenuation processes. The mixing of sediments with limestone filler could be applied to the remediation of zones affected by mining activities, because the toxicological effect on the tested organisms in the stabilised sediments was reduced significantly and the metal(loid) content was diminished.     

Inheritance and heritability of resistance to citrus leprosis

ABSTRACT The genetic inheritance of resistance to leprosis, the most important viral disease of citrus in Brazil, was characterized through the phenotypic assessment of 143 hybrids resulting from crosses between tangor 'Murcott' (Citrus sinensis x C. reticulata) and sweet orange 'Pêra' (C. sinensis), considered to be resistant and susceptible to the disease, respectively. All plants were grafted onto Rangpur lime (C. limonia) and inoculated with Citrus leprosis virus, cytoplasmic type through the infestation with viruliferous mites, Brevipalpus phoenicis. The experiments were arranged in a completely randomized block design with 10 replicates. Incidence and severity of the disease in leaves and stems as well as plant growth parameters (plant height and stem diameter) were recorded for 3 years after the infestation with the viruliferous mites. The average values of all variables were analyzed using principal component analysis, discriminant factorial analysis, estimation of the clonal repeatability coefficients, and frequency of the distributions of the average values for each measured variable. The principal component analysis resulted in the identification of at least two groups with resistance and susceptibility to leprosis, respectively. About 99% of all hybrids were correctly classified according to the discriminant factorial analysis. The broad-sense heritability coefficients for characteristics associated with incidence and severity of leprosis ranged from 0.88 to 0.96. The data suggest that the inheritance of resistance to leprosis may be controlled by only a few genes.