E12 technique: Conventional epoxy resin sheet plastination

Epoxy plastination techniques were developed to obtain thin transparent body slices with high anatomical detail. This is facilitated because the plastinated tissue is transparent and the topography of the anatomical structures well preserved. For this reason, thin epoxy slices are currently used for research purposes in both macroscopic and microscopic studies. The protocol for the conventional epoxy technique (E12) follows the main steps of plastination—specimen preparation, dehydration, impregnation and curing/casting. Preparation begins with selection of the specimen, followed by freezing and slicing. Either fresh or fixed (embalmed) tissue is suitable for epoxy plastination, while slice thickness is kept between 1.5 and 3 mm. Impregnation mixture is made of epoxy E12 resin plus E1 hardener (100 ppw; 28 ppw). This mixture is reactive and temperature sensitive, and for this reason, total impregnation time under vacuum at room laboratory temperature should not last for more than 20–24 hr. Casting of impregnated slices is done in either flat chambers or by the so‐called sandwich method in either fresh mixture or the one used for impregnation. Curing is completed at 40°C to allow a complete polymerization of the epoxy‐mixture. After curing, slices can be photographed, scanned or used for anatomical study under screen negatoscope, magnification glass or fluorescent microscope. Based on epoxy sheet plastination, many anatomical papers have recent observations of and/or clarification of anatomical concepts in different areas of medical expertice.     

DNA hybridization assay for detection of nucleopolyhedrovirus in whitemarked tussock moth (Orgyia leucostigma) larvae

DNA dot‐blot hybridization assays utilizing a horseradish peroxidase‐labelled whole genomic DNA probe and enhanced chemiluminescence were conducted to quantify detection thresholds of nucleopolyhedrovirus (NPV) in whitemarked tussock moth (Orgyia leucostigma) larvae. The minimum detection thresholds for an aqueous suspension of occlusion bodies (OBs), OBs added to macerates of non‐infected larvae and OBs in macerates of diseased larvae were 7.8 × 10³, 7.8 × 10³, and 1.5 × 10³ OBs, respectively. Purified viral DNA was detected at a concentration of 1.6 × 10⁻¹ ng in a 20 µl volume. The presence of pre‐occluded viral nucleocapsids and DNA, inherent to infected larvae, improved the detection threshold five‐fold compared with OBs alone. Larval tissues did not block the detection system utilized, nor did they bind non‐specifically to the probe. Detection thresholds, upon sequential hybridization of the same membrane, on average deteriorated two‐fold between the first and second hybridization and an additional six‐fold between the second and third hybridization. NPV infection was detected two days post‐inoculation (pi) in about one‐third of the larvae examined and in almost all larvae three days pi. Microscopic analysis of stained larval smears missed NPV infection in almost all larvae two days pi and about two‐thirds of the larvae three days pi. Results from the two methods of analysis were not comparable until four days pi. The detection system utilized is a reliable, efficient and simple method for the early detection of NPV infection in large numbers of larvae and may be used for further studies quantifying the role of this baculovirus in the ecology of whitemarked tussock moth populations. © 2001 Society of Chemical Industry     

Genetics of nitrate content in lettuce

Summary Adverse effects on human health makes the high nitrate content frequently found in lettuce (Lactuca sativa L.) grown under low light conditions an undesirable trait. Efforts have been made to breed cultivars with a reduced capacity for nitrate accumulation. In this study components of variance for nitrate content were estimated in F₂ and F₃ generations of ten lettuce crosses. Additive genotypic variances (A) were estimated from F₃ variance components and from the covariance between F₂ plants and corresponding F₃ lines. Estimates of wide sense heritability of the F₂ from crosses between a high nitrate genotype and four low nitrate genotypes ranged from 0.44 to 0.74 and the estimates for A ranged from 0.25 to 0.40 g·l^-1. Estimated wide sense heritabilities of F₂'s from six crosses involving two low nitrate parents ranged from 0.15 to 0.52. The parents of four of the low nitrate crosses showed relatively large effects of genotype x environment (GE) interaction in successive experiments: the nitrate content of the parents reacted differently to environmental changes between experiments. Estimates of A for crosses between low nitrate genotypes without large effects of GE interaction ranged from 0 to 0.19 g·l^-1. The estimated probability of selecting transgressive low nitrate lines in the progeny of a cross between a high and a low nitrate genotype was low (P=0.002–0.039), indicating that large populations should be evaluated to combine the positive traits of modern high nitrate cultivars with low nitrate content from genotypes not adapted to modern cropping practices. In the progenies from crosses between two low nitrate genotypes without important GE effects, only low estimates of the probability of obtaining transgressive low nitrate lines were obtained (P=0.04–0.06). With the growth conditions used in this study, the probability of selecting lines with a nitrate content compatible under all winter conditions with the proposed future maximum permissible level of 2.5 g nitrate per kg fresh matter is low. Therefore the solution of this problem should be found in a combination of low nitrate cultivars and cultural measures that reduce the nitrate content of the crop.     

Grassland conversion along a climate gradient in northwest China: Implications for soil carbon and nutrients

Soil organic carbon (SOC) stocks and nutrient availability are key indicators of soil quality, and both can be influenced by land-use change. However, it is still unclear whether the impact of land-use change on SOC and nutrient stocks differs between ecoregions. Grasslands near the northeast border of the Qinghai-Tibetan Plateau (QTP) occur across several ecoregions that have recently been subjected to substantial land-use change. Based on long-term land-use history, we conducted a field investigation comparing soil C and nutrient stocks between natural grassland (NGL) and three types of converted grassland (agricultural grassland, AGL; farmland, FL; and abandoned farmland, AFL) in three ecoregions along a climate gradient: alpine meadow, temperate steppe and temperate desert. Compared with NGL, soil C stocks in converted grasslands were 22%–30% lower in the alpine meadow, but 60–82% higher in the temperate steppe and 6%–76% higher in the temperate desert. Converted grasslands also contained higher stocks of available nitrogen and phosphorus than NGL in the temperate steppe and desert. Soils (0–40 cm) in NGL contained 14.8 ± 0.1 kg C m⁻² in alpine meadow, 6.7 ± 0.6 kg C m⁻² in temperate steppe and 1.7 ± 0.3 kg C m⁻² in temperate desert. Together, our results indicate that the responses of soil C and nutrients to grassland conversion differed between ecoregions. Thus, to optimize soil C sequestration rates and overall soil quality, we suggest that land-use policies in this area should take into account local environmental conditions.     

Isolation of bacteria from fermented food and grass carp intestine and their efficiencies in improving nutrient value of soybean meal in solid state fermentation

Background: Soybean meal is an excellent and cost-effective protein source; however, its usage is limited in the piglet due to the presence of anti-nutritional factors and the antigens glycinin and β-conglycinin. The objective of the current study was to screen and select for bacteria that can be efficiently adopted to ferment soybean meal in order to solve this problem.Results: Bacteria were isolated from fermented soy foods and the grass carp intestine, and strains selected for high protease, cellulase and amylase activities. The isolated bacteria were characterized as Bacillus cereus, Bacillus subtilis and Bacilus amyloliquefacien, respectively. Fermentation with food-derived Isolate-2 and fish-derived F-9 increased crude protein content by 5.32% and 8.27%, respectively; improved the amino acid profile by increasing certain essential amino acids, broke down larger soy protein to 35 k Da and under, eliminated antigenicity against glycinin and β-conglycinin, and removed raffinose and stachyose in the soybean meal following a 24-h fermentation.Conclusions: Our results suggest these two B. amyloliquefaciens bacteria can efficiently solid state ferment soybean meal and ultimately produce a more utilizable food source for growing healthy piglets.     

External quality assurance system for antibiotic resistance in bacteria of animal origin in Europe (ARBAO-II), 2003

Initiated in 2003 by the European Union, ARBAO-II aims to establish a monitoring of antimicrobial susceptibility among the veterinary laboratories in all European countries based on validated methodologies. This includes an external quality control system for the most important bacterial pathogens. In 2003 two trials were performed. The first on Salmonella and Escherichia coli involved 31 laboratories in 18 countries and the second on Staphylococcus and Streptococcus included 22 laboratories in 15 countries. For the E. coli strains, 92.8% of the results complied with the reference MICs, for Salmonella 93.7%, for Streptococcus 80.4% and for Staphylococcus 93.1%. Most problems were observed when testing florfenicol (79.2% correct), gentamicin (84.2%) and amoxicillin + cl (84.9%) in E. coli; streptomycin in Salmonella (62.5%); gentamicin (56.7%), lincomycin (71.4%), clindamycin (75.4%), TMP + sulfonamides (75.7%) and chloramphenicol (78.5%) in Streptococcus; erythromycin (81.5%) and oxacillin (78.2.5%) in Staphylococcus. A few laboratories caused most deviations. However, there was no correlation between good performance for one bacterial group and good performance for other groups. This study showed that most laboratories are capable of performing correct susceptibility testing for E. coli and Salmonella, even though performance of some laboratories can be improved, and that some problems exist for Staphylococcus and Streptococcus. This clearly shows the need for continuous harmonisation of methodologies within the EU.     

Prevalence and characteristics of quinolone resistance in Escherichia coli in veal calves

Quinolone resistance is studied and reported increasingly in isolates from humans, food-producing animals and companion animals. Resistance can be caused by chromosomal mutations in topoisomerase genes, plasmid-mediated resistance genes, and active transport through efflux pumps. Cross sectional data on quinolone resistance mechanisms in non-pathogenic bacteria from healthy veal calves is limited. The purpose of this study was to determine the prevalence and characteristics of quinolone resistance mechanisms in Escherichia coli isolates from veal calves, after more than 20 years of quinolone usage in veal calves. MIC values were determined for all isolates collected as part of a national surveillance program on antimicrobial resistance in commensal bacteria in food-producing animals in The Netherlands. From the strains collected from veal calves in 2007 (n=175) all isolates with ciprofloxacin MIC ≥ 0.125 mg/L (n=25) were selected for this study, and screened for the presence of known quinolone resistance determinants. In this selection only chromosomal mutations in the topoisomerase type II and IV genes were detected. The number of mutations found per isolate correlated with an increasing ciprofloxacin MIC. No plasmid-mediated quinolone resistance genes were found. The contribution of efflux pumps varied from no contribution to a 16-fold increase in susceptibility. No correlation was found with the presence of resistance genes of other antimicrobial classes, even though all quinolone non-wild type isolates were resistant to 3 or more classes of antibiotics other than quinolones. Over twenty years of quinolone usage in veal calves in The Netherlands did not result in a widespread occurrence of plasmid-mediated quinolone resistance, limiting the transmission of quinolone resistance to clonal distribution.     

COMPUTED TOMOGRAPHY OF CHOLESTERINIC GRANLOMAS IN THE CHOROID PLEXUS OF HORSES

Cholesterinic granulomas are tumor-like masses in the choroid plexuses of horses. This report describes the computed tomographic findings in four horses with cholesterinic granulomas, including the clinical and the pathologic features. All four horses had bilateral cholesterinic granulomas in the lateral ventricles. Computed tomographic images clearly delineated the masses, the opacity of the granulomas, and the enhancement pattern after intravenous bolus injection of contrast medium; these factors varied considerably. Additional CT findings were symmetric or asymmetric widening of the lateral ventricles.     

Genotype × Environment Interaction for Nitrate Concentration in Lettuce

High nitrate concentration is a problem when lettuce (Lactuca sativa L.) is grown under low light intensities. Genotypes with low nitrate concentrations have been identified and are being used in breeding programmes. Previous results indicated the occurrence of genotype × environment (GE) interactions. Because of the important influence of light intensity on nitrate accumulation, two types of interactions may be expected: interactions related to daily variation, and those related to annual variation. In the present investigation both types were investigated using eight butterhead genotypes of lettuce which were repeatedly harvested. No daily variation in nitrate concentration and no corresponding GE-interactions were found, irrespective of the level of global radiation. In contrast, a large annual variation and important corresponding GE-interactions were found. Joint regression analysis on environmental means and on physical factors related to light intensity showed a differential response of genotypes to changing environmental conditions. Multiple joint regression on daylength and change in daylength accounted for two-thirds of the interaction variance. However, deviations from regression were still significant indicating non-linearity of the relationship, or, the existence of other environmental factors contributing to GE-interaction.