Antibodies to bovine serum albumin in swine sera: implications for false-positive reactions in the serodiagnosis of African swine fever

Antibodies to bovine serum albumin were detected in swine sera by use of an immunoblotting technique. Such sera had false-positive reactions, as determined by results of African swine fever virus serodiagnostic techniques when bovine serum albumin was a contaminant in the soluble cytoplasmic antigen obtained from infected cells cultured in the presence of bovine serum. The soluble cytoplasmic antigen obtained from cell cultures infected with African swine fever virus in the presence of porcine serum did not react with the false-positive sera and, therefore, was used for African swine fever virus serodiagnostic methods, with 0% false-positive results.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Detection of African swine fever virus antibodies by immunoblotting assay.

An immunoblotting assay has been adapted to detect antibodies against African swine fever virus. The electrophoretic transfer of proteins and the immunoreaction conditions were optimized, using 4 mA/cm2 of current intensity and 10 micrograms of soluble cytoplasmic antigen of infected cells per strip. Filters of polyvinylidene difluoride showed the highest capacity for protein absorption, but nitrocellulose filters showed lower backgrounds. The specificity and the pattern of the proteins induced by African swine fever virus that react with the antisera were determined in immunoblotting assay, IP30 being the most reactive protein.     

Hematology and serum chemistry values of the European brown hare

Background: Hematology and serum biochemistry values can be useful for evaluating the health of populations of captive and free-living European brown hares ( Lepus europaeus ). However, current information is fragmented and incomplete.
Objective: The objective of this study was to determine normal hematology and serum chemistry values for a population of captive European brown hares and to investigate differences related to sex and age.
Methods: Routine hematology and serum chemistry values were determined for 48 captive European brown hares in Catalonia (northeastern Spain): 21 young (6 males and 15 females) and 27 adult (14 males and 13 females) hares. Blood was obtained by auricular venipuncture while animals were physically restrained. Results were expressed as the mean ± SD, and reference intervals were calculated as the mean ± 2SD.
Results: RBC count, PCV, concentrations of hemoglobin, total protein, albumin, gamma globulin, urea nitrogen, and creatinine, and alanine aminotransferase activity were significantly higher in adults than in young hares, and alkaline phosphatase activity and total bilirubin concentration were lower. Sex differences were not observed.
Conclusions: Hematology and serum chemistry results obtained in this study are considered representative for healthy captive European brown hares. A single reference interval can be used for values from male and female hares, however, age-specific intervals are needed for some analytes.     

Seed dormancy and germination in the summer annual Galeopsis speciosa

This study examined germination and dormancy in Galeopsis speciosa (Lamiaceae), a common summer annual weed in cold‐temperate areas. Seeds collected in southern Sweden were subjected to several experiments. The seeds were dormant at maturity. Seeds sown outdoors after collection produced a small number of seedlings that emerged early in the spring. After long cold stratification or stratification outdoors over two winters, the maximum germination was 40–50%; germination occurring over a wide range of temperatures. Warm stratification preceding cold stratification had no effect on germination, but repeated warm and cold periods seemed to promote germination. Gibberellic acid (GA) stimulated germination, but full germination was only achieved after more than 2 months of incubation at the most suitable temperature regime tested. Excised embryos grew and developed into normal seedlings. With these results, the species does not fit into the currently used system for seed dormancy classifications. The response to GA and the growth of excised embryos indicate non‐deep or intermediate physiological dormancy, but dormancy alleviation by stratification was not in line with the guiding principles for these classifications. Galeopsis speciosa has a strong dormancy that is sufficiently alleviated during the winter to allow germination of only part of a seed batch each year; hence a stepwise germination pattern occurs over a period of several years.     

Cryopreservation of Ram Semen in Extenders Containing Soybean Lecithin as Cryoprotectant and Hyaluronic Acid as Antioxidant

A soybean lecithin‐based extender supplemented with hyaluronic acid (HA) was assayed for effectiveness to improve the quality of frozen–thawed ram semen. HA has not been tested yet in an extender containing soybean lecithin for freezing ram semen. Thus, the aim of this study was to analyse the effects of soybean lecithin at 1% or 1.5% along with HA at 0, 0.5 and 1 mg ml^‐1 in a Tris‐based extender on the motion characteristics, membrane integrity (HOST), viability, GSH peroxidase (GSH‐PX) activity, lipid peroxidation and acrosomal status after freezing–thawing. Semen was collected from four Mehraban rams during the breeding season and frozen in the six lecithin×HA extenders. The extender containing 1.5% lecithin supplemented with no HA yielded higher total motility (52.5%±1.6), viability (55.8%±1.6) and membrane integrity (44.5%±1.7), but the effects of the lecithin concentration did not reach signification. Linearity‐related parameters, ALH, BCF, lipid peroxidation, GSH‐PX activity, morphology and acrosomal status were not affected by the extender composition. In general, adding HA significantly decreased sperm velocity (1 mg ml^‐1 HA), total motility (only with 1.5% lecithin), viability (1 mg ml^‐1 HA for 1% lecithin; both concentrations for 1.5% lecithin) and membrane integrity. In conclusion, adding HA to the freezing extender supplemented with soybean lecithin failed to improve quality‐related variables in ram semen. Increasing the lecithin content could have a positive effect, but further studies are needed.     

Severe hematometra in a dog with cystic endometrial hyperplasia/pyometra complex.

An 18-month-old, intact female German shepherd dog was presented for evaluation of severe, frank hemorrhage from the vulva. A complete blood count, serum biochemical analysis, and urinalysis supported acute hemorrhage. Abdominal radiographs were unremarkable. Abdominal ultrasound demonstrated that the uterine body and cranial vaginal lumen were fluid-filled and contained hyperechoic, polypoid masses that were suspected to be blood clots. Exploratory laparotomy revealed a mildly enlarged uterus with a diffuse, cystic endometrium; significant amounts of blood and blood clots within the lumen; and several focal accumulations of yellow fluid presumed to be a purulent exudate. Histopathology confirmed severe, cystic, endometrial hyperplasia and pyometra complex.     

Nitrogen availability in some Wisconsin forests: comparisons of resin bags and on-site incubations

Summary Estimates of ammonium and nitrate availability in conifer and hardwood forests using an ion exchange resin (IER) bag method and with on-site incubations of soil cores in buried bags were compared. Correlations between the two methods were generally high. Correlation coefficients (r) between IER nitrate and buried-bag mineralized nitrate ranged from 0.87 to 0.92. Both methods also correlated well with aboveground net primary production, litter fall N content, and fine root biomass. The major differences between the methods related to the relative importances of ammonium and nitrate forms of available N. The IER method indicated that both ammonium and nitrate were important on all sites, with nitrate predominating in most soils. The buried-bag results indicated that available N was primarily in the form of nitrate (all ammonium was oxidized), but that nitrate was insignificant on infertile sites.