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.     

First results of embryonic development,spawning and larval rearing of the Mediterranean spider crab Maja squinado (Herbst ) under laboratory conditions,a candidate species for a restocking program

The Mediterranean spider crab, Maja squinado, is depleted due to overfishing. The crab has virtually disappeared from areas where it was abundant, such as the Balearic Islands and the Catalan coast. Maja squinado, is economically and ecologically very valuable, and it is essential to obtain information on its biology and rearing conditions to attempt to repopulate the damaged stocks of the species in the Mediterranean basin. Herein, we describe the first successful rearing of M. squinado under laboratory conditions. Our results show that M. squinado is an excellent candidate for restocking using cultured juveniles. Two consecutive broods with a 1–4 day interbrood period were observed in the laboratory in wild‐caught females, the maximum observed duration of embryonic development of the egg mass being 32 days at 18.4 ± 0.9°C, and went through four different stages. The complete larval and first juvenile development was studied in laboratory cultures fed enriched Artemia metanauplius. At 19.6 ± 0.6°C, development from hatching to first crab moult took 17 days, and it comprised two zoeae stages and one megalopa stage. The survival rate at the different stages was monitored, and 7.13 ± 2.3% was achieved at the first crab instar.     

Searching the most appropriate sample pretreatment for the elemental analysis of wines by inductively coupled plasma-based techniques

Different sample preparation methods were evaluated for the simultaneous multielement analysis of wine samples by inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS). Microwave-assisted digestion in closed vessel, thermal digestion in open reactor, and direct sample dilution were considered for the determination of Li, Be, Na, Mg, Al, K, Ca, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Sr, Y, Mo, Cd, Ba, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, Tl, Pb, and Bi in 12 samples of red wine from Valencia and Utiel-Requena protected designation of origin. ICP-MS allows the determination of 17 elements in most of the samples, and using ICP-OES, a maximum of 15 elements were determined. On comparing the sample pretreatment methodology, it can be concluded that the three assayed procedures provide comparable results for the concentration of Li, Na, Mg, Al, K, Ca, Mn, Fe, Zn, and Sr by ICP-OES. Furthermore, ICP-MS data found for Cu, Pb, and Ba were comparable. Digestion treatment provides comparable values using both total decomposition in open system and microwave-assisted treatment for Cu by ICP-OES and for Cr, Ni, and Zn by ICP-MS. Open vessel total digestion provides excess values for Cr, Mn, Fe, and Zn by ICP-OES and defect values for Se. However, direct measurement of diluted wine samples provided uncomparable results with the digestion treatment for Mn, Cu, Pb, Zn, Ba, and Bi by ICP-OES and for Mg, Cr, Fe, Ni, and Zn by ICP-MS. Therefore, it can be concluded that microwave-assisted digestion is the pretreatment procedure of choice for elemental analysis of wine by ICP-based techniques.     

Night-time Melatonin Secretion and Seasonally Delayed Puberty in Gilts

This study was conducted to determine whether the seasonal delay in puberty in autumn is driven by individual differences in night-time melatonin secretion in domestic gilts at the attainment of puberty. A group of spring-born gilts (n = 30) were expected to reach puberty in autumn by the age of 7 months. Eighteen of these gilts were selected in pairs on the basis of matched days of birth. By the expected time, half of the animals showed oestrous symptoms (group CYCLING, n = 9) with the rest remaining silent (group SILENT, n = 9). Afterwards, all gilts were fitted with indwelling jugular catheters for frequent blood sampling. Blood samples were collected from all animals three times during the day followed by three times in the night at 2-h intervals for 48 h. The samples were analysed by a commercial radioimmunoassay (RIA). The results show a consistent 25-fold rise (on average) in night-time melatonin concentration in every animal sampled with group averages ranging from 0.28 +/- 0.04 to 0.37 +/- 0.06 pg/ml at day and from 10.20 +/- 2.16 to 10.67 +/- 0.05 pg/ml at night. Night-time group mean values between CYCLING and SILENT gilts did not differ significantly (10.26 +/- 0.67 and 10.38 +/- 0.94 for the CYCLING; 10.67 +/- 0.05 and 10.20 +/- 2.16 for the SILENT). When 10 pg/ml was used as a threshold value, six individuals did not reach it during the night (low responders). Two of these gilts were CYCLING and four were SILENT. In conclusion, the results presented imply no involvement of the level of night-time melatonin concentration in the seasonal delay of puberty in gilts.     

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.     

Factors Affecting Fertility in Loosely Housed Sows and Gilts: Vulvar Discharge Syndrome, Environment and Acute-phase Proteins

The effect of vulvar discharge syndrome (VDS) on sow and gilt fertility was studied on 26 farms. Of 824 animals inspected in 21 randomly selected and five VDS problem farms, 19 (2.3%) were afflicted with VDS. Altogether 542/799 of the examined animals (67.8%) farrowed thereafter. Nine of the 19 VDS animals (47.4%) and 533/780 non-VDS animals (68.3%) farrowed at the first chance after the examination (p = 0.05). None of the unmated gilts in this study had VDS. Environmental and individual factors likely to be associated with fertility and VDS were tested. In multivariate analysis, factors associated with farrowing were VDS, reproductive status, availability of roughage and confinement to individual stalls. None of the variables tested was associated with VDS. However, all of the VDS problem farms were overcrowded and had concrete, partly slatted floors with little or no bedding. The median value of haptoglobin (Hp) was 2.5 (range 1.3-3.1) g/l in VDS animals and 2.3 (0.5-4.3) g/l in controls (p = 0.6). The median C-reactive protein (CRP) in VDS animals was 30.3 (3.3-171.3) mg/l and in controls 25.9 (3.3-361.1) mg/l (p = 0.7). In conclusion, VDS decreased fertility of gilts and sows in the absence of a systemic acute-phase response, as indicated by stable concentrations of Hp and CRP.     

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.