Development and application of an enzyme-linked immunosorbent assay for detection of bovine viral diarrhea antibody based on Erns glycoprotein expressed in a baculovirus system.

The BVDV envelope glycoprotein E(rns)/gp48 and the C terminal 79 amino acids of the capsid protein coding region were expressed in a baculovirus system and antigenically characterized. Western blot assay was used to detect recombinant E(rns) (r-E(rns)) in infected insect cells using specific monoclonal antibodies. The r-E(rns) was then used in an indirect ELISA to detect BVDV specific antibodies in a panel of 540 well-characterized sera. Results of the r-E(rns) ELISA were compared to those obtained with a commercially available competitive ELISA targeting anti-NS2/3 antibodies. A good correlation was observed between the 2 ELISA (kappa = 0.916, 95% C.I.: 0.876, 0.956). Using the commercial NS2/3 ELISA as the reference test, the relative sensitivity of r-E(rns) ELISA was 97.5% (95% C.I.: 94.3%, 99.1%) and the relative specificity was 93.9% (95% C.I.: 89.4%, 96.9%), while relative specificity was 100% (95% C.I.: 97%, 100%) using true negative sera (derived from a negative herd). All but 1 antigen positive animals (n = 36) tested negative in the r-E(rns) ELISA; among them all 22 confirmed PI animals were negative by r-E(rns) ELISA. The ability of r-E(rns) ELISA to identify cattle immunized with inactivated vaccine was also demonstrated in a small group of cattle, compared to an NS2/3 antibody ELISA. Results suggest that r-E(rns) ELISA represents an alternative test for antibody generated by natural infection or BVDV vaccination.     

The pond's shape matters: differential growth,physiological condition and survival of epibenthic Farfantepenaeus aztecus postlarvae

As a function of the water quality provided by square, circular and oval experimental ponds, the growth, survival and oxygen requirements in epibenthic postlarvae of Farfantepenaeus aztecus were analysed in relation to their routine metabolism and apparent heat increment. Temperature, oxygen concentration, pH and salinity were measured daily in two experimental ponds of each shape. The postlarvae oxygen consumption during two 24‐h cycles, their growth, physiological condition and survival and the productivity in the ponds were estimated. Low values of pH, oxygen concentration and phytobenthos productivity, and reduced postlarvae relative growth and survival were observed in the square ponds. We suggest that the latter results from a deficient water circulation related to the effect of the pond's shape on dissolved oxygen levels and, consequently, on growth and survival. The postlarvae routine metabolism, including feeding, varied between 1.91 and 2.25 mg O₂ h^?1 g^?1 wet weight, whereas the minimum oxygen concentration needed in the ponds is approximately 4.25 mg O₂ L^?1. These conditions were achieved in the oval ponds concurrent with higher survival and growth values, in which individuals distributed randomly, for which we suggest that oval‐shaped ponds could be the most adequate for the culture of this and other penaeid species.     

HPLC separation and NMR structural elucidation of sn-1,2-, 2,3-, and 1,3-diacylglycerols from olive oil as naphthylethylurethane derivatives

In this study, sn-1,2-, sn-2,3-, and sn-1,3-diacylglycerols were isolated from olive oil, and their urethane derivatives (urethanes) were prepared. Normal-phase high-performance liquid chromatography (NP-HPLC) separation of the urethane isomers was performed and the separate classes were studied by nuclear magnetic resonance (NMR). The use of 1H NMR and homo- and heteronuclear 2D techniques provided a great amount of information in a very short time, particularly when a high-field NMR instrument (700 MHz) was used. Particularly diagnostic for this kind of compound was the glyceridic moiety that presents typical chemical shifts both for carbon and hydrogen. These studies show the usefulness of NMR spectroscopy to recognize clearly the sn-1,3- and, moreover, sn-1,2- with respect to sn-2,3-diacylglycerols, although very minor differences occur between them.     

The use of medetomidine-based sedation protocols to perform urohydropropulsion and cystoscopy in the dog

Voiding urohydropropulsion and cystoscopy are routine procedures performed in the dog for diagnostic and therapeutic purposes. These procedures are typically performed under general anesthesia. The purpose of this study was to describe the use of medetomidine-based sedation protocols to perform voiding urohydropropulsion and cystoscopy in cardiovascularly healthy, non-diabetic dogs without evidence of urinary obstruction, renal disease, or hepatic disease. Results of this study revealed significantly shorter procedure times and decreased cost in sedated dogs, with diagnostic and therapeutic outcomes equivalent to those of patients that underwent general anesthesia. Based on the results of this retrospective study, the authors recommend medetomidine-based sedation protocols for voiding urohydropropulsion and cystoscopy in appropriately selected patients.     

Effect of triploidy on quality traits of shi drum (Umbrina cirrosa L.) until the second rearing year

Carcass and flesh morphometric, reological and chemical traits of triploid (3n) and diploid (2n) shi drum (Umbrina cirrosa L.) were evaluated through 7 months. Three age groups, 17‐, 21‐ and 24‐month‐old fish, were investigated. The effects of ploidy were statistically evaluated and the weight of fish was included in the model as a covariate because triploids grew less than diploids. As expected, fish weight was found to be significantly correlated with all the investigated morphometric traits, but showed a negative correlation with some chemical (pH) and colour traits (lightness) of raw fillet. In comparison with controls, triploid shi drums were characterized by different morphological traits that involved a slender body shape. In triploids, a reduction in condition factor, backbone weight, dressing index and an increase in the agility index were also recorded. When the commercial size (i.e. over 300 g) was achieved, triploids exhibited larger coelomatic and fillet (dorsal white muscle) fat deposition than diploids. Among reological traits, colour and texture were affected by ploidy; raw fillet lightness and cooked flesh tenderness were higher among triploids in all the investigated age groups. As fish were fed with a restricted feeding regimen, fillet fat deposition was supposed to be limited. Thus, the effects of ploidy on reological traits may be only partially explained by lipid fillet amount and are supposed to be more related to different fibre muscle architecture.     

Effect of accelerated autolysis of yeast on the composition and foaming properties of sparkling wines elaborated by a champenoise method

Five mutants (obtained by UV mutagenesis) and the parent strain were selected to produce sparkling wines following the traditional or champenoise method. The wines were aged with the yeast for 9 months, with samples being taken each month for analytical and sensory determinations. The wines elaborated with mutant strain IFI473I demonstrated an accelerated release of protein, amino acids, and polysaccharides. An analysis of the secreted polysaccharides revealed that mannose was the major sugar present. The effects of the products released by yeasts on the foaming properties of the wines were determined by both sensory and instrumental analysis. In all cases, the wines elaborated with mutant strain IFI473I showed improved foaming properties as compared to wines fermented without this strain. Similar results were obtained at a decreased aging time of 6 months, thereby confirming the capacity of IFI473I strain to carry out an accelerated autolysis. These results demonstrate that mutant strain IFI473I can significantly reduce production times of high-quality sparkling wines.     

Activating mutations of NOTCH1 in human T cell acute lymphoblastic leukemia

Very rare cases of human T cell acute lymphoblastic leukemia (T-ALL) harbor chromosomal translocations that involve NOTCH1, a gene encoding a transmembrane receptor that regulates normal T cell development. Here, we report that more than 50% of human T-ALLs, including tumors from all major molecular oncogenic subtypes, have activating mutations that involve the extracellular heterodimerization domain and/or the C-terminal PEST domain of NOTCH1. These findings greatly expand the role of activated NOTCH1 in the molecular pathogenesis of human T-ALL and provide a strong rationale for targeted therapies that interfere with NOTCH signaling.     

Genetic and antigenic characterization of the matrix protein of two genetically distinct ovine lentiviruses

Small Ruminant Lentiviruses (SRLV) are a group of non-oncogenic retroviruses including Maedi-Visna virus (MVV) and Caprine Arthritis-Encephalitis virus (CAEV), which cause a chronic, multisystemic disease in sheep and goats, respectively. Phylogenetic analyses of SRLV are based in most cases on partial pol sequences. Several reports indicate that the species specificity of these viruses is not as strict as previously thought; MVV-like viruses have been found in goat populations and vice versa. Recently, the sequencing of some Italian ovine isolates has shown the presence of a new cluster more similar to classical caprine isolates (CAEV-like). Few data are available on the variability of structural proteins involved in the antibody response of infected animals. In this study, the gag gene of two genetically distinct ovine isolates, namely the MVV-like It-561 and the CAEV-like It-Pi1, was sequenced and the epitopes of matrix protein (MA) were mapped. Recombinant MAs and their subunits from both ovine aforementioned strains were tested against a panel of sheep and goat sera. Reactive epitopes were found in all three subunits of MA, although the central subunit displayed a more consistent reactivity. Epitope mapping of this subunit demonstrated that the amino acid sequence of at least one immunodominant epitope was quite different in the two strains. This antigenic variability may affect the sensitivity of a single strain-based immunoassay and suggests that both SRLV genotypes should be used in the development of future diagnostic tests, to avoid viral strain selection during the eradication programmes.     

Genetic and antigenic characterization of the surface lipoprotein P48 of Mycoplasma bovis

The presence of a membrane lipoprotein homologous to the P48 of Mycoplasma agalactiae was investigated in different Mycoplasma bovis isolates selected by geographical locations and biological properties. Its potential as a diagnostic tool was also discussed. The presence of a specific signal observed in all M. bovis field isolates probed with a rabbit antiserum raised against the M. agalactiae recombinant P48 demonstrated that this protein is structurally and antigenically conserved within the M. bovis cluster. No signal was detected when testing six different mycoplasma species found in cattle. The p48 gene was identified by PCR approach and partially sequenced. Full length gene sequence was obtained by direct bacterial chromosome sequencing. Five UGAs were selectively mutated into UGG and the full length mutated gene, lacking the signal peptide, was cloned and expressed in Escherichia coli. The purified recombinant antigen (r-P48) was evaluated as a potential marker of infection using a panel of 86 well-characterized sera from experimentally and naturally infected cattle. Specific IgM antibodies were detected within 6-9 days after experimental infection followed by an IgG response lasting from the third/fourth week after contact. Although antibody titers were well below those observed in sheep or goats infected with M. agalactiae, results suggest that M. bovis r-P48 can be used as a specific marker of infection.