Multiresidue assay for benzimidazole anthelmintics by liquid chromatography and confirmation by gas chromatography/selected-ion monitoring electron impact mass spectrometry

A multiresidue method utilizing all-disposable labware has been developed for 8 benzimidazole anthelmintics from ovine, bovine, and swine muscle and liver tissues. After an initial extraction with ethyl acetate and subsequent evaporation, a 3-component extraction using hexane, ethanol, and 0.2N HCl was used for final cleanup. Clean extracts were produced for separation and determination by reverse-phase liquid chromatography at 298 nm, using methanol and aqueous buffer as mobile phase. A synthesized internal standard, 2-(n-butylmercapto)benzimidazole, was used for quantitation of all drugs. Results are included along with statistical information verifying the performance of the method. Spiked control tissues and incurred drug tissues were used for an intralaboratory study with a concentration range of 50-1470 ppb. A series of standard curves at 0, 50, 100, and 200 ppb were analyzed. Overall recovery at the 100 ppb level averaged 92% (CV 8%) in liver tissues, across all 3 species and 88% (CV 5%) in muscle tissues across all 3 species. Results were confirmed by gas chromatography/mass spectrometry with acid hydrolysis of the remaining extract in 2N HCl followed by re-extraction of the amine and derivatization to the tert-butyldimethylsilyl derivative. The anthelmintics were identified by gas chromatography/selected ion monitoring electron-impact mass spectrometry. Ion ratio measurements were taken and compared to standard material. CVs averaged 10% or less for all drugs tested.     

猪圆环病毒病疫苗免疫母猪的生产现场试验

本文介绍了一系列的试验结果,证实一种新的猪Ⅱ型圆环病毒(PCV-2)疫苗可免疫母猪以减少母猪和新生仔猪潜在的圆环病毒病的排毒,因而可减少仔猪出生后早期的病毒暴露量,并通过母源抗体保护仔猪,以防止PCV-2的早期感染。     

Effect of storage duration on egg quality,embryo mortality and hatchability in FUNAAB-ɑ chickens

Effect of extended storage on egg quality, embryo mortality and hatchability in FUNAAB-ɑ chickens was determined. Hatchable eggs (n = 288; weighing 53.2 ± 4.67 g) collected from a flock of FUNAAB-ɑ layer breeder hens aged 32 weeks were stored in egg tray with broad end up under 16 ± 1.5°C for either 0, 4, 8, 12, 16 or 20 d. Before incubation, eight eggs from each group were evaluated for internal and external quality traits. Remaining eggs were set in an incubator and transferred into hatcher on embryonic day 18. Data collected were subjected to one-way analysis of variance. Egg weight loss (EWL; p < .001), surface area (p < .001), yolk diameter (p < .001), inner and outer blastoderm diameters (p < .05) and dead in germ (DIG; p < .001) increased with storage duration while yolk height (p < .001), yolk index (p < .001), albumen weight (p < .05), albumen height (p < .05), albumen index (p < .01), Haugh's unit (HU; p < .05), fertility (p < .001), hatchability of set (HATCHS; p < .001) and fertile eggs (p < .05) decreased. Weight losses of 0, 1.2, 2.2, 3.4, 4.6 and 6.1% were recorded in egg stored for 0, 4, 8, 12, 16 and 20 days respectively. Eggs stored beyond 8 days exhibited higher DIG and lower HATCHS. Shell percentage in 4 days storage (11.4%) was lower (p < .05) than in 16 days storage (13.4%). Shell thickness was similar in eggs stored for 0 to 12 days, but 8 days storage (0.60 mm) had thinner (p < .01) shell than day 16 (0.71 mm) and day 20 (0.73 mm) storage. Internal quality unit (IQU) was higher (p < .05) in fresh eggs (180.4) than in 12 days (167.8) and 20 days (167.8) stored eggs. Extended storage of FUNAAB-ɑ eggs caused EWL, surface area shrinkage, lowered HU and IQU, loss of yolk and albumen quality, increased blastoderm diameters and DIG, and decreased egg fertility and HATCHS from day 8 forward. Storing FUNAAB-ɑ eggs beyond 8 days reduced quality parameters; therefore, other mitigating factors are recommended when storing beyond 8 days.     

Selective silencing of foreign DNA with low GC content by the H-NS protein in Salmonella

Horizontal gene transfer plays a major role in microbial evolution. However, newly acquired sequences can decrease fitness unless integrated into preexisting regulatory networks. We found that the histone-like nucleoid structuring protein (H-NS) selectively silences horizontally acquired genes by targeting sequences with GC content lower than the resident genome. Mutations in hns are lethal in Salmonella unless accompanied by compensatory mutations in other regulatory loci. Thus, H-NS provides a previously unrecognized mechanism of bacterial defense against foreign DNA, enabling the acquisition of DNA from exogenous sources while avoiding detrimental consequences from unregulated expression of newly acquired genes. Characteristic GC/AT ratios of bacterial genomes may facilitate discrimination between a cell's own DNA and foreign DNA.     

Effects of additive iron on growth,tissue distribution,haematology and immunology of gilthead sea bream, <Emphasis Type="Italic">Sparus aurata</Emphasis>

The comparative effects of iron-supplemented levels on growth, tissue distribution, haematology and immunology of gilthead sea bream, Sparus aurata (2 g) were investigated, using four organic (50, 100, 200, 300 mg ORG/kg diet) and one inorganic iron source (200 INOR mg/kg diet). Fish were treated for 12 weeks with the experimental diets and maintained at a water temperature of 19–22°C. Growth (final weight and specific growth rate), tissue distribution (spleen, liver and muscle), haematological parameters (red blood cells, haematocrit, haemoglobin and mean corpuscular haemoglobin concentration) and non-specific immune indexes (respiratory burst activity and antibacterial activity of serum) were analysed. No significant differences were found in growth and iron tissue distribution among the tested groups. Red blood cell counting was statistically higher in fish given 50 ORG, 100 ORG, 200 ORG and 200 INOR feeds. However, haematocrit, haemoglobin and mean corpuscular haemoglobin concentration were not significantly affected by increasing dietary iron. Fish receiving the 100 ORG diet had the best performance with respect to the respiratory burst activity and significantly higher values for antibacterial activity of serum were obtained in fish fed with the 300 ORG diet. The present findings provided no clear evidence of the optimum iron concentration. However, there was adequate indication that iron supplementation enhanced the performance of gilthead sea bream, mainly from a haematological and immunological point of view.     

Determination of oil,emulsifier and polymer or salt content of spray adjuvants

A method for the separation of the oil, emulsifier and polymer or salt fractions of adjuvants is described. The adjuvant is dissolved in 90% aqueous ethanol and filtered to remove polymers or salt. Light petroleum (distillation range 30–40°C) is added to the solution in a separating funnel. The light petroleum fraction is subsequently extracted with successive aliquots of 80, 70, 60 and 50% aqueous ethanol. The whole of the ethanol and light petroleum fractions are evaporated to dryness to yield the emulsifier and oil fractions respectively. Emulsifier, oil and polymer or salt are determined gravimetrically. The method successfully detected the presence of polymer or salt in adjuvants, but could not distinguish between them when both were present in the same product. The adjuvants could be classified into four broad classes; those containing > 70% emulsifier (wetter-spreaders), those containing > 75% oil (anti-evaporants), those containing a high proportion of polymer (film-forming substances), and those containing comparable amounts of all three components (multi-purpose adjuvants).