Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.     

Manipulated Mouse Embryos as Bioassay System for Water Quality Control

Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K⁺ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system.     

ACE inhibition in goats under fixed‐time artificial insemination protocol increases the pregnancy rate and twin births

To assess the effect of the angiotensin‐converting enzyme (ACE) inhibition on the efficiency of the fixed‐time artificial insemination (TAI), 69 goats were divided randomly into two groups: enalapril (n = 35) and control (n = 34). In the experiment, all animals underwent the protocol of fixed‐time artificial insemination for 12 days. Enalapril group received enalapril maleate dissolved in saline (Enalapril, Lab Teuto Ltda) subcutaneously at the following doses: 0.2 mg/kg/day in D0‐D2; 0.3 mg/kg/day in D3‐D6 and 0.4 mg/kg/day in D7‐D11. The control group received the corresponding volume of 0.9% saline solution. We performed a single insemination 36 hr after sponge removal using frozen semen from two adult male goats with recognized fertility. The ultrasound pregnancy diagnosis was 30 days after the artificial insemination (AI). There was significant increase in pregnancy rates and twinning as well as a decrease in foetal loss in animals receiving enalapril (p < .01). The use of ACE inhibitors during the TAI protocol was shown to be a promising alternative to increase the efficiency of such reproductive biotechnology.     

Biology and life table of Diaeretiella rapae against Brevicoryne brassicae on different cultivars of cauliflower

Phytoparasitica - Diaeretiella rapae (McIntosh) (Hymenoptera: Braconidae) is one of the most promising, effective and a common endoparasitoid of different aphid species including cabbage aphid,...     

To pool or not to pool? Guidelines for pooling samples for use in surveillance testing of infectious diseases in aquatic animals

Samples from multiple animals may be pooled and tested to reduce costs of surveillance for infectious agents in aquatic animal populations. The primary advantage of pooling is increased population‐level coverage when prevalence is low (<10%) and the number of tests is fixed, because of increased likelihood of including target analyte from at least one infected animal in a tested pool. Important questions and a priori design considerations need to be addressed. Unfortunately, pooling recommendations in disease‐specific chapters of the 2018 OIE Aquatic Manual are incomplete and, except for amphibian chytrid fungus, are not supported by peer‐reviewed research. A systematic review identified only 12 peer‐reviewed aquatic diagnostic accuracy and surveillance studies using pooled samples. No clear patterns for pooling methods and characteristics were evident across reviewed studies, although most authors agreed there is a negative effect on detection. Therefore, our purpose was to review pooling procedures used in published aquatic infectious disease research, present evidence‐based guidelines, and provide simulated data examples for white spot syndrome virus in shrimp. A decision tree of pooling guidelines was developed for use by peer‐reviewed journals and research institutions for the design, statistical analysis and reporting of comparative accuracy studies of individual and pooled tests for surveillance purposes.     

Cell wall polysaccharides from chalkumra (Benincasa hispida) fruit. Part I. Isolation and characterization of pectins

Pectic polysaccharides were obtained from chalkumra (Benincasa hispida) fruit by sequential extraction with ammonium oxalate (fraction BOX), dilute acid (fraction BHCl), and cold dilute alkali (fraction BOH). The highest yield of polysaccharides was obtained with oxalate and HCl. BOX was enriched in partly methyl-esterified galacturonic acid, whereas BHCl and BOH contained mostly galactose. All of the extracts showed similar elution patterns in size exclusion chromatography although the intrinsic viscosities (eta) were different (132 +/- 6, 100 +/- 5, and 285 + 10 mL/g for BOX, BHCl, and BOH, respectively). From fractionation by anion exchange chromatography, homogalacturonan (as seen from sugar analysis and Fourier transform infrared spectrum) accounted for more than half of BOX and 11% of BHCl. Methylation analyses and hydrolysis of BHCl with endo-beta-(1-->4)-d-galactanase showed the presence of beta-(1-->4)-d-galactan. The neutral galactan represented more than 76% of BHCl and approximately 40% of BOH. The other polysaccharides were complex galactans in BOH and an acidic arabinan (<1%) in BOX and BHCl.     

Comparative Performance of Rice with System of Rice Intensification (SRI) and Conventional Management using Different Plant Spacings

The System of Rice Intensification (SRI) reportedly enhances the yields of rice (Oryza sativa L.) through synergy among several agronomic management practices. This study was conducted to investigate the effects on rice plant characteristics and yield by comparing the plants grown with different methods of cultivation – SRI vs. recommended management practices (RMP) focusing on the impact of different plant spacings. Performance of individual hills was significantly improved with wider spacing compared with closer‐spaced hills in terms of root growth and xylem exudation rates, leaf number and leaf sizes, canopy angle, tiller and panicle number, panicle length and grain number per panicle, grain filling and 1000‐grain weight and straw weight, irrespective of whether SRI or RMP was employed. Both sets of practices gave their highest grain yield with the spacing of 20 × 20 cm; however, SRI yielded 40 % more than the recommended practice. At this spacing, canopies also had the highest leaf area index (LAI) and light interception during flowering stage. The lowest yield was recorded at 30 × 30 cm spacing under both the practices, as a result of less plant population (11 m^?2), despite improved hill performance. During the ripening stage, hills with wider spacing had larger root dry weight, produced greater xylem exudates, and transported these towards shoot at faster rates. These features contributed to the maintenance of higher chlorophyll levels, enhanced fluorescence and photosynthesis rates of leaves and supported more favourable yield attributes and grain yield in individual hills than in closely‐spaced plants. Moreover, these parameters further improved in SRI, apart from the enhanced percentage of effective tillers and showed substantial and positive impacts on grain yield (17 %) compared with recommended practice. In conclusion, wide spacing beyond optimum plant density, however, does not give higher grain yield on an area basis and for achieving this, a combination of improved hills with optimum plant population must be worked out for SRI.     

Potential of a novel antibiotic, 2-methylheptyl isonicotinate, as a biocontrol agent against fusarial wilt of crucifers

Screening for newer bioactive compounds from microbial metabolites resulted in the isolation of a novel antibiotic from the culture filtrate, Streptomyces sp 201, of a soil. The bioactive compound, with antifungal and antibacterial activity, was identified as 2-methylheptyl isonicotinate. The antifungal activity of live culture, culture broth and the isolated bioactive compound showed marked inhibition against dominant soil-borne phytopathogens such as Fusarium oxysporum Schlect, F moniliforme Sheldon, F semitectum Berkeley & Ravenel, F solani (Martius) Sacc and Rhizoctonia solani Kuehn. The compound had no effect on seed germination and seedling development as displayed by root and stem growth of the test plant species. In pot experiments with seedlings of cruciferous plants such as Raphanus sativus L (radish), Brassica campestris L (yellow mustard), Brassica oleracea var botrytis L (cauliflower), the antibiotic compound showed promising protective activity of 92% when seeds of the test plants were treated at a dose of 50 micrograms ml-1 prior to sowing. Seed treatment with a spore suspension (3 x 10(8) spores ml-1) of the Streptomyces sp 201 displayed protective activity in the range of 56-60%. Seeds coated with 2.5% methyl cellulose-amended spores of the antagonist showed protective activity in the range of 64-72%. Further, seed treatment with the culture filtrate of the antagonist also showed promising protective activity in the range of 64-84%.     

Gonadotropin‐induced Puberty Does Not Impair Reproductive Performance of Gilts over Three Parities

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty‐nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post‐weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning‐to‐oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.     

Molecular markers based genetic relatedness studies in tissue culture propagated japanese plum cultivars Santa Rosa and Frontier

Genetic Resources and Crop Evolution - Santa Rosa and Frontier are the major Japanese plum (Prunus salicina Lindl.) cultivars grown throughout the world. The present investigation was performed to...