Quantitation of adenine nucleotides in equine colonic mucosal tissue using high performance liquid chromatography.

The objectives were to use high performance liquid chromatography (HPLC) to validate an established method for adenine nucleotide separation in equine colonic mucosal tissue, to determine the inherent variability in the tissue and extraction method, and to determine the stability of ATP, ADP, and AMP in the tissue with time. Equine colonic mucosal tissue obtained from a single horse was immediately submersed in liquid nitrogen, and stored at -70 degrees C. Samples were lyophilized, extracted, and separated by HPLC. The limit of quantitation was 0.05 microg/mL. The coefficient of variation for the instrument was less than 10% for all nucleotides measured. When the tissue was not homogenized prior to sampling, there were significant differences in adenine nucleotide content between samples. However, when the tissue was homogenized prior to analysis, these differences were no longer significant. There was no significant decrease in ATP, ADP, or AMP content over a 54-day analysis period.     

几种因素对培养小鼠精原干细胞的影响

探讨了小鼠胚胎成纤维细胞饲养层及干细胞因子(SCF)、白血病抑制因子(LIF)、碱性成纤维细胞生长因子(bFGF)等对体外培养小鼠精原干细胞的作用.结果显示,在饲养层上,精原干细胞贴壁时间缩至8～12 h,饲养层具有促进其分裂增殖的作用;培养液中加入不同浓度的SCF、LIF及bFGF,可延长精原干细胞在体外的存活时间,其中加入30μg/LSCF后,其体外存活时间与对照组相比差异显著(P<0.05).     

Transmission of porcine reproductive and respiratory syndrome virus from persistently infected sows to contact controls.

The objective of this study was to determine if porcine reproductive and respiratory syndrome virus (PRRSV) could persist in non-pregnant sows and if persistently infected sows could transmit virus to naive contact controls. Twelve PRRSV-naive, non-pregnant sows (index sows) were infected with a field isolate of PRRSV and housed in individual isolation rooms for 42 to 56 days postinfection. Following this period, 1 naive contact sow was placed in each room divided by a gate allowing nose-to-nose contact with a single index sow. Index sows were not viremic at the time of contact sow entry. Virus nucleic acid was detected by polymerase chain reaction, and infectious virus was detected by virus isolation in sera from 3 of the 12 contact sows at 49, 56, and 86 days postinfection. All 3 infected contacts developed PRRSV antibodies. Virus nucleic acid was detected in tissues of all of the 12 index sows at 72 or 86 days postinfection. Nucleic acid sequencing indicated that representative samples from index and infected contacts were homologous (> 99%) to the PRRSV used to infect index sows at the onset of the study. This study demonstrates that PRRSV can persist in sows and that persistently infected sows can transmit virus to naive contact animals.     

Teratogenicity testing of BI 58 EC (38% dimethoate) in chicken embryos with special respect to degradation of the active ingredient.

The insecticide formulation BI 58 EC was tested for teratogenicity in chicken embryos, with particular reference to degradation of the active ingredient (dimethoate) after the treatment of embryonated eggs. The pesticide was diluted in water to a concentration level of 0.8%, and the emulsion was injected into the air space in a volume of 0.1 ml/egg, or hen's eggs were treated by the immersion technique. Residues of dimethoate were measured in the samples on days, 13, 15 and 19 of the incubation of chicken embryos, and morphological examinations were performed simultaneously. Analytical chemistry data indicated a slower degradation of dimethoate in embryos after the immersion of eggs, and cyllosis was remarkable in this group among the sporadic developmental anomalies. The liver tissues of both treated groups exhibited severe fatty infiltration.     

Research note: Postnatal development of electrolyte transport in calf rumen as affected by weaning time.

In a previous study we found a positive correlation between early weaning in calves and morphological parameters which were indicative of ruminal development, i.e. the length and width of the papillae. The objective of the present study was to determine to what extent this observation could be reflected by modulations of absorptive and secretory functions of the rumen mucosa. For this purpose the short-circuit currents (Isc) as a measure of electrogenic net ion fluxes and the transepithelial conductances (G(T)) as a measure of the overall tissue permeability were measured in vitro applying the Ussing-chamber technique. Simultaneously, the unidirectional flux rates of sodium and chloride across rumen wall epithelia were determined in the absence of electrochemical gradients. Under these conditions, significant positive net flux rates (Jnet) clearly indicate active mechanisms for electrolyte absorption. For the experiments 12 male Holstein calves 7 d of age were assigned to three groups of 4 animals each: milk group (I, slaughtered after 6 weeks of age), late weaning group (II, slaughtered after 9 weeks of age) and early weaning group (II, weaned after 6 weeks of age and slaughtered after 9 weeks of age). Whereas G(T) values remained unaffected by different age and feeding, Isc values were significantly affected by early weaning but were not influenced by age. Irrespective of weaning time active absorption of Na+ tended to be higher by about 60% in 9 weeks old animals. Active absorption of chloride was significantly increased in milk fed 9 weeke old calves and this effect was further stimulated by early weaning. In conclusion, the data show an increasing active Na+ absorption with age in calf rumen that could not be influenced by early weaning. Similarly, active Cl- absorption was initially increased during postnatal development and this effect could be stimulated further by early weaning.     

Rabies virus infection in Aedes pseudoscutellaris cells: a study on receptorial structures

Rabies virus is able to infect in vitro a wide range of homeothermic and poikilothermic cells but little is known about its multiplication in arthropod cells. In this research the infection of rabies virus in Aedes pseudoscutellaris cells, a mosquito cell line susceptible to mosquito-borne viruses, was studied. After 60 days of incubation at 26 degrees C up to 70% of infected cells showed the synthesis of both viral nucleocapsid and envelope antigens, although viral yield and cell damage could not be detected. Research performed in order to investigate the role of membrane carbohydrate moieties in rabies virus-mosquito cell interaction suggested the participation of galactose and N-acetylglucosamine whereas sialic acid, known to be a rabies binding site in many homeothermic cell lines, was not involved.     

Mitigation of N2O emissions from grassland by nitrification inhibitor and Actilith F2 applied with fertilizer and cattle slurry

Abstract. Nitrous oxide (N₂O) is involved in both ozone destruction and global warming. In agricultural soils it is produced by nitrification and denitrification mainly after fertilization. Nitrification inhibitors have been proposed as one of the management tools for the reduction of the potential hazards of fertilizer-derived N₂O. Addition of nitrification inhibitors to fertilizers maintains soil N in ammonium form, thereby gaseous N losses by nitrification and denitrification are less likely to occur and there is increased N utilization by the sward. We present a study aimed to evaluate the effectiveness of the nitrification inhibitor dicyandiamide (DCD) and of the slurry additive Actilith F2 on N₂O emissions following application of calcium ammonium nitrate or cattle slurry to a mixed clover/ryegrass sward in the Basque Country. The results indicate that large differences in N₂O emission occur depending on fertilizer type and the presence or absence of a nitrification inhibitor. There is considerable scope for immediate reduction of emissions by applying DCD with calcium ammonium nitrate or cattle slurry. DCD, applied at 25 kg ha^–1, reduced the amount of N lost as N₂O by 60% and 42% when applied with cattle slurry and calcium ammonium nitrate, respectively. Actilith F2 did not reduce N₂O emissions and it produced a long lasting mineralization of previously immobilized added N.     

The Effect of Diet on the Biochemical Composition of Juvenile Artemia: Potential Formulations for Rock Lobster Aquaculture

The lipid class and fatty acid (FA) composition of juvenile Artemia fed continuously on four diets—the microalga Tetraselmis suecica , a mix of oat bran-wheat germ-lecithin (OWL), OwL-eicosapentaenoic acid (EPA), and OWL-EPA-arachidonic acid (AA)—were examined over a 9-d experiment in an attempt to approximate the FA profile of phyllosoma larvae of wild southern rock lobster Jasus edwardrii . The main difference in lipid class composition of Artemia fed the four diets was the relative level of polar lipid (PL) and triacylglycerol (TAG). By day 9, the algal-fed Artemia were highest in PL (95% of total lipid) and lowest in TAG (2%), whereas the remaining diets resulted in Artemia with 16–30% PL and 41–82% TAG. After 2 d, the relative FA composition of all Artemia treatments closely reflected those of the diets, with no marked change after further feeding (to day 9). In terms of the content of essential polyunsaturated fatty acids (PUFA), by day 5 Artemia fed: 1) with the algal diet contained 7 mg/g FA dry mass (0.3% DHA, 6.3% EPA, 3.4% AA of total FA); 2) with the OWL diet contained 3 mg/g (0.3% DHA, 0.9% EPA, 0.7% AA); 3) with the OWL-EPA diet contained 55 mg/g (6.2% DHA, 11.6% EPA, 1.1% AA); and 4) with the OWL-EPA-AA contained 83 mg/g (3.8% DHA, 7.5% EPA, 17.4% AA). The PUFA profiles of Artemia using the OWL-oil diets were similar to wild rock lobster phyllmmata, although levels of doco-sahexaenoic acid (DHA) were lower (10% DHA) than in J. edwardsii larvae. On the basis of PUFA composition data alone, the results suggest the suitability of the OWL-oil mixed diets for consideration for feeding to Artemia used in the culture of southern rock lobster larvae, particularly if the level of DHA can be further enhanced.