In vitro maturation and fertilization of prepubertal and pubertal black Bengal goat oocytes

Oocytes retrieval, in vitro maturation (IVM) and fertilization (IVF) efficiency are inevitable steps towards in vitro production of embryos. In the present study, these parameters were investigated in the ovaries of prepubertal (n = 31) and pubertal (n = 61) black Bengal goats obtained from a slaughterhouse. Nuclear maturation was evaluated upon aspiration and following IVM in TCM-199 (Earle''s salt with L-glutamine and sodium bicarbonate) for 27 h at 39℃ under 5% CO₂ in humidified air. The oocytes retrieval and efficiency (mean ± SD) per prepubertal and pubertal goats were 5.2 ± 0.6 and 6.8 ± 0.6, and 77.3 ± 0.1% and 80.5 ± 0.6%, respectively. Anaphase I - telophase I stages differed significantly (7.3 ± 0.8 vs. 2.6 ± 0.2, p < 0.05) between the two groups of goats. After IVM, the percentages of metaphase II were significantly higher (66.3 vs. 60.3, p < 0.05) in pubertal goats than in their prepubertal counterparts. The percentages of normal in vitro fertilization (IVF) in Fert-Tyrode''s albumin lactate pyruvate of pubertal goat oocytes did not differ between Percoll and swim-up sperm separation methods (36.7 ± 0.9% vs. 32.7 ± 1.3%, p > 0.05). Furthermore, sperm capacitation by heparin alone or in combination with ionomycin did not lead to a significant increase in the normal fertilization rate (34.8 ± 1.7 vs. 32.2 ± 1.5%, respectively) in the oocytes of pubertal goats. In conclusion, the ovaries of pubertal black Bengal goats obtained from the slaughterhouse could be used for in vitro embryo production. However, further optimization of the IVM and IVF techniques are necessary for satisfactory in vitro embryo production.     

Development of on-line measurement system of bulk density based on on-line measured draught, depth and soil moisture content

On-line measurement of soil compaction is needed for site specific tillage management. The soil bulk density (ρ) indicating soil compaction was measured on-line by means of a developed compaction sensor system that comprised several sensors for on-line measurement of the draught (D) of a soil cutting tool (subsoiler), the soil cutting depth (d) and the soil moisture content (w). The subsoiler D was measured with a single shear beam load cell, whereas d was measured with a wheel gauge that consisted of a swinging arm metal wheel and a linear variable differential transducer (LVDT). The soil w was measured with a near infrared fibre-type spectrophotometer sensor. These on-line three measured parameters were used to calculate ρ, by utilising a hybrid numerical–statistical mathematical model developed in a previous study. Punctual kriging was performed using the variogram estimation and spatial prediction with error (VESPER) 1.6 software to develop the field maps of ρ, soil w, subsoiler d and D, based on 10 m × 10 m grid. To verify the on-line measured ρ map, this map was compared with the map measured by the conventional core sampling method.

The spherical semivariogram models, providing the best fit for all properties was used for kriging of different maps. Maps developed showed that no clear correlation could be detected between different parameters measured and subsoiler D. However, the D value was smaller at shallow penetration d, whereas large D coincided with large ρ values at few positions in the field. Maps of ρ measured with the core sampling and on-line methods were similar, with correlation coefficient (r) and the standard error values of 0.75 and 0.054 Mg m⁻³, respectively. On-line measured ρ exhibited larger errors at very dry zones. The normal distribution of the ρ error between the two different measurement methods showed that about 72% of the errors were less than 0.05 Mg m⁻³ in absolute values. However, the overall mean error of on-line measured ρ was of a small value of 2.3%, which ensures the method accuracy for on-line measurement of ρ. Measurement under very dry conditions should be minimised, because it can lead to a relatively large error, and hence, compacted zones at dry zones cannot be detected correctly.     

Extended glucuronidation is another major path of cyanidin 3-O-beta-D-glucopyranoside metabolism in rats

We previously determined that five rather hydrophobic metabolites appeared in blood plasma after oral administration of cyanidin 3-O-beta-D-glucopyranoside, but a group of hydrophilic metabolites still remained unidentified. In the present study, 12 hydrophilic metabolites found were collected from urine and plasma samples by high-performance liquid chromatography (HPLC) and then analyzed by tandem MS spectrometry. From the MS spectra, four metabolites out of 12 were assigned as glucuronides of cyanidin 3-O-beta-D-glucopyranoside and six out of 12 were glucuronides of the primary metabolites of cyanidin 3-O-beta-D-glucopyranoside (O-methyl cyanidin 3-O-beta-D-glucopyranoside). Extended glucuronides of cyanidin 3-O-beta-D-glucopyranoside and O-methyl cyanidin 3-O-beta-D-glucopyranoside showed their maximum plasma concentrations at 15 and 60 min (or 30 min) after oral administration, respectively. Their maximum plasma concentrations ranged from 15 to 70 nM. From the profile of urinary-excreted anthocyanins after intravenous administration, it was deduced that extended glucuronides of cyanidin 3-O-beta-D-glucopyranoside and O-methyl cyanidin 3-O-beta-D-glucopyranoside were mainly produced in the liver rather than by intestinal flora. The area under the plasma concentration curve was 0.25 micromol min/L for extended glucuronides of cyanidin 3-O-beta-D-glucopyranoside and 0.14 micromol min/L for O-methyl cyanidin 3-O-beta-D-glucopyranoside, respectively, when evaluated as cyanidin 3-O-beta-D-glucopyranoside equivalent, indicating that extended glucuronidation is a critical pathway in cyanidin 3-O-beta-D-glucopyranoside metabolism in rats.     

Earthworm casts form stable aggregates in physically degraded soils

Topsoils affected by surface mining suffer severe physical degradation and lose most of their earthworm populations. After mining, replaced soils are planted to grassland and managed to improve soil structure. Earthworm inoculation into selected restored areas produced populations similar to those of undisturbed soils within 3 years. Soil properties in inoculated areas were compared with those of controls to evaluate the contribution of casts to bulk soil aggregation, and soil organic matter and root content responses to earthworm activity. Crumb porosity and coarse particle content were measured in water-stable macro-aggregates and earthworm casts to establish whether aggregates were formed by earthworms. Over a 5- to 6-year period, inoculation increased stable aggregation (>2 μm, >60 μm and >3 mm), even at 0- to 5-cm depth where it reduced soil organic matter content. Productivity and root content were also increased by inoculation; roots and organic matter were re-distributed to greater depth. Crumb porosity decreased with casts > aggregates (inoculated plots) > aggregates (control plots). Coarse particle content increased with casts < aggregates (inoculated plots) < aggregates (control plots). Coarse particle and porosity data were consistent with much of the newly aggregated soil being processed and formed by earthworms as casts. Whilst levels of soil organic matter were often closely associated with percentage stable aggregation, root content showed weaker associations. Aggregation percentage was most closely associated with abundance of Aporrectodea longa, although at particular depths significant correlations were also obtained for Aporrectodea caliginosa and Lumbricus terrestris. Results suggest that earthworms, rather than plant roots, initiate aggregation in severely degraded grassland soils.     

Development and Characterization of Novel Polymorphic Microsatellite Markers for Tapinoma indicum (Hymenoptera: Formicidae)

Tapinoma indicum (Forel) (Hymenoptera: Formicidae) is a nuisance pest in Asia countries. However, studies on T. indicum are limited, especially in the field of molecular biology, to investigate the species characteristic at the molecular level. This paper aims to provide valuable genetic markers as tools with which to study the T. indicum population. In this study, a total of 143,998 microsatellite markers were developed based on the 2.61 × 10⁶ microsatellites isolated from T. indicum genomic DNA sequences. Fifty selected microsatellite markers were amplified with varying numbers of alleles ranging from 0 to 19. Seven out of fifty microsatellite markers were characterized for polymorphism with the Hardy–Weinberg equilibrium (HWE) and linkage disequilibrium (LD) analysis. All seven microsatellite markers demonstrated a high polymorphic information content (PIC) value ranging from 0.87 to 0.93, with a mean value of 0.90. There is no evidence of scoring errors caused by stutter peaks, no large allele dropout, and no linkage disequilibrium among the seven loci; although loci Ti-Tr04, Ti-Tr09, Ti-Te04, Ti-Te13, and Ti-Pe5 showed signs of null alleles and deviation from the HWE due to excessive homozygosity. In conclusion, a significant amount of microsatellite markers was developed from the data set of next-generation sequencing, and seven of microsatellite markers were validated as informative genetic markers that can be utilized to study the T. indicum population.     

A greenhouse method for screening lettuce for resistance to Sclerotinia sclerotiorum

Agar blocks colonized by Sclerotinia sclerotiorum, causal agent of lettuce drop, were used as inoculum in screening for resistance to this pathogen in the greenhouse.Since the degree of resistance to the pathogen among cultivars varied with the time observed after inoculation, resistance was assessed by the number of days for 50% of the plants of each cultivar to die, as well as the number of plants that died. The cultivars ‘PI (Plant Introduction) 250427’, ‘Taiwan’, ‘PI 251790’, ‘PI 255568’, ‘MSU 73-44’ and ‘Bibb’ were found to have a higher level of resistance to S. sclerotiorum than the cultivar ‘Grand Rapids’ at 12 or 15 days following inoculation.     

Seasonal variations in the numbers of trichostrongylid nematode eggs and their larvae in the faeces of farmed goats in Malaysia.

Faecal worm egg counts of goats from two farms in Penang Island, West Malaysia, were monitored over a period of 14 months. The faecal egg count pattern followed that of total rainfall. The humid tropical environment was favourable for the development of various species of trichostrongylid nematodes, namely Haemonchus contortus, Trichostrongylus spp., Oesophagostomum spp. and Cooperia spp. Generally, H. contortus was observed to be the predominant species, more so in the monsoon months of the year.