Molecular characterization of reticuloendotheliosis virus insertions in the genome of field and vaccine strains of fowl poxvirus

Evidence of the widespread occurrence of reticuloendotheliosis virus (REV) sequence insertions in fowl poxvirus (FPV) genome of field isolates and vaccine strains has increased in recent years. However, only those strains carrying a near intact REV provirus are more likely to cause problems in the field. Detection of the intact provirus or REV protein expression from FPV stocks has proven to be technically difficult. The objective of the present study was to evaluate current and newly developed REV and FPV polymerase chain reaction (PCR) assays to detect the presence of REV provirus in FPV samples. The second objective was to characterize REV insertions among recent "variant" FPV field isolates and vaccine strains. With REV, FPV, and heterologous REV-FPV primers, five FPV field isolates and four commercial vaccines were analyzed by PCR and nucleotide sequence analysis. Intact and truncated REV 5' long terminal repeat (LTR) sequences were detected in all FPV field isolates and vaccine strains, indicating heterogeneous REV genome populations. However only truncated 3' LTR and envelope sequences were detected among field isolates and in one vaccine strain. Amplifications of the REV envelope and 3' LTR provided strong evidence to indicate that these isolates carry a near intact REV genome. Three of the four FPV vaccine strains analyzed carried a solo complete or truncated 5' LTR sequence, indicating that intact REV provirus was not present. Comparison of PCR assays indicated that assays amplifying REV envelope and REV 3' LTR sequences provided a more accurate assessment of REV provirus than PCR assays that amplify the REV 5' LTR region. Therefore, to differentiate FPV strains that carry intact REV provirus from those that carry solo 5' LTR sequences, positive PCR results with primers that amplify the 5' LTR should be confirmed with more specific PCR assays, such as the envelope, or the REV 3' LTR PCR.     

Estimates of Breed Average Direct,Maternal and Heterosis Effects for Some Pre-weaning Traits of Zebu Breeds and Their Crosses in Southeastern Mexico

Information on 936 birth weights (BW), adjusted weaning weights (AWW) and average daily gains (ADG) from Brahman (B), Indubrazil (I), Gyr (G) and commercial Zebu (C) cattle and their crosses were obtained from the records of a farm in the State of Yucatan, Mexico. Two statistical models were used to analyse the data: the first included the effects of year and season of birth, sex of the calf, cow's parity number and breed group. The other model included the same environmental effects as the previous model and the genetic components of breed group (breed additive, breed maternal and heterosis effects). Additive and maternal effects were expressed as deviations from those for the B breed. The means of BW, AWW and ADG for the B breed were 33.3 ± 0.64 kg, 204.0 ± 4.9 kg and 760.06 ± 19.24 g/day, respectively. Both models gave similar results in terms of the variance explained. Additive effects were significant (p < 0.05) for AWW and ADG but not for BW. The minor additive effect was for the C breed (−10.75 kg and −40.77 g for WW and ADG, respectively). Maternal and heterosis effects were not significant (p > 0.05) for any of the traits. The correlation between the breed group means and the means estimated by the prediction equation was r = 0.88. Finally, there was no significant genetic effect on pre-weaning calf performance among the genotypes evaluated. The genetic model can be used to predict the genotype means with a high degree of accuracy. This revised version was published online in July 2006 with corrections to the Cover Date.     

Quantitative changes in the normal and apoptotic thymocytes of pigs treated with anabolic doses of the beta2 adrenergic agonist clenbuterol

Although the beta2 adrenergic agonists have been seen to have important effects on the mechanisms regulating the development and death of T-cells in the thymus, the side-effects on the immune system of anabolic treatments of these substances have hardly been considered. In order to evaluate the effects exerted by the beta2 adrenergic agonist clenbuterol on the thymocyte population, the thymus of eight pigs treated with anabolic doses of this substance was studied by morphometric methods, regarding apoptotic (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive) and normal (TUNEL-negative) cells. The thymus of another eight pigs fed without clenbuterol served as a control. The clenbuterol treatment had a clear effect on the thymocyte size, decreasing their mean nuclear area. The T-cell apoptosis index was also affected by the clenbuterol, significantly increasing the apoptosis percentage in the treated group with respect to the control. In the light of our results, the clenbuterol induced thymocyte apoptosis throughout the thymus and caused morphometric changes in the thymocyte population, which was in line with the immunosuppressive role attributed to other beta2 adrenergic agonists.     

The effects of GnRH and adrenergic agents on PRL and beta-endorphin secretion by porcine pituitary cells in vitro

The direct effects of alpha- and beta-adrenergic agents on PRL and beta-endorphin (beta-END) secretion in vitro by porcine pituitary cells have been investigated. Pituitary glands were obtained from mature gilts, which were ovariectomised (OVX) one month before slaughter. Ovariectomised gilts, assigned to four groups, were primed with: (1) vehicle (OVX); (2) and (3) oestradiol benzoate (EB; 2.5 mg/100 kg b.w.) at 30-36 h (OVX+EB I) and 60-66 h (OVX+EB II) before slaughter, respectively; and (4) progesterone (P4; 120 mg/100 kg b.w.) for 5 consecutive days before slaughter (OVX+P4). Isolated anterior pituitary cells were submitted to 3.5 h incubation in the presence of GnRH, alpha- and beta-adrenergic agonists [phenylephrine (PHEN) and isoproterenol (ISOP), respectively], or alpha- and beta-adrenergic blockers [phentolamine (PHENT) and propranolol (PROP), respectively]. The culture media were assayed for PRL (exp. I) and beta-endorphin-like immunoreactivity (beta-END-LI) (experiment II). In experiment I, GnRH did not influence PRL release by pituitary cells in all experimental groups. Some of tested doses of adrenergic agonists, PHEN and ISOP, increased PRL release from pituitary cells of OVX gilts, but not from those of OVX+EB I animals. In the OVX+EB II group, PHEN alone, but ISOP with PROP, potentiated PRL secretion by the cells. In OVX+P4 animals, PHEN alone or in combination with PHENT and also ISOP alone or with PROP enhanced PRL output from the cells. In experiment II, addition of GnRH increased beta-END-LI release from pituitary cells only in the OVX+EB II group. PHEN and PHENT potentiated beta-END-LI secretion by pituitary cells in OVX+EB II and OVX+P4 groups, while ISOP and PROP increased beta-END-LI secretion by the cells of OVX and OVX+EB II animals. In turn, in the OVX+EB I group, effect of PHENT and PROP on PRL secretion by pituitary cells was inhibitory. In conclusion, our results suggest that adrenergic agents can modulate PRL and beta-END secretion by porcine pituitary cells in a manner dependent on the hormonal status of gilts.     

Effect of two thermal regimes on the muscle growth dynamics of sea bass larvae,Dicentrarchus labrax L

Muscle growth was studied in larvae of sea bass, Dicentrarchus labrax L., reared at two temperatures: real ambient temperature ( congruent with 15 degrees C during vitelline phase and increased gradually) and 19 degrees C from fertilization until the end of larval development. Muscle cellularity, body length and body weight were measured. Early temperature influenced larval development and so, pre-larval phase finished earlier at 19 degrees C than at ambient temperature (4 and 6 days, respectively). Temperature also affected muscle growth such that at hatching and at mouth opening hypertrophy of muscle fibres was greater at 19 degrees C (P < 0.05), whereas hyperplasia was similar in both groups. After 25 days, the cross-sectional area of the white muscle was greater at 19 degrees C (P < 0.05), which was mainly associated with a higher proliferation of new white muscle fibres. At this stage the body length was also higher at 19 degrees C. Metamorphosis finished earlier in fish reared at 19 degrees C (52 days) than at natural temperature (82 days). At this developmental stage body length and cross-sectional area of the myotome were similar in both groups. However, muscle cellularity differed between groups. Thus, hypertrophy of muscle fibres was higher in fish reared at ambient temperature (P < 0.05), whereas proliferation of new muscle fibres was higher at 19 degrees C (P > 0.05).     

Antioxidant activity of coffee model systems

Coffee model systems prepared from combinations of chlorogenic acid (CGA), N(alpha)-acetyl-1-arginine (A), sucrose (S), and cellulose (C) were roasted at 240 degrees C for 4 min prior to analysis by UV-visible spectrophotometry, capillary zone electrophoresis (CZE), and the ABTS radical cation decolorization assay. The A/CGA/S/C and A/S/C systems were also fractionated by gel filtration chromatography. Antioxidant activity of the systems showed a positive, nonlinear relationship with the amount of CGA remaining after roasting. Sucrose degradation was a major source of color in the heated systems. There was no relationship between antioxidant activity and color generation.     

Oral administration of yeast, Saccharomyces cerevisiae, enhances the cellular innate immune response of gilthead seabream (Sparus aurata L.)

The effects of including lyophilised whole yeast, Saccharomyces cerevisiae, in the diet on the seabream innate immune response were investigated. Gilthead seabream (Sparus aurata L.) specimens were fed four different diets for 4 weeks: a commercial diet as control and the same diet supplemented with 1, 5 or 10 g/kg yeast. After 1, 2 and 4 weeks, serum complement titres, as a humoral parameter, and phagocytic, respiratory burst, myeloperoxidase and natural cytotoxic activities of head-kidney leucocytes, as cellular parameters, were evaluated. The results showed that yeast supplements enhanced all the latter responses, but not the humoral response. This enhancement was dose-dependent except for the cytotoxic activity that was only stimulated by the lower dose of yeast assayed. As yeast cell walls are able to enhance the seabream cellular innate immune response, these results support the possible use of whole yeast as natural inmunostimulants in common fish diets.     

Dorsal laminectomy for caudal cervical spondylomyelopathy: postoperative recovery and long-term follow-up in 20 dogs

OBJECTIVE: To evaluate the postoperative morbidity and long-term outcome of dogs after dorsal laminectomy for caudal cervical spondylomyelopathy (CCSM). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Twenty dogs with CCSM. METHODS: Medical records of dogs treated by dorsal laminectomy for CCSM at North Carolina State University and Colorado State University between 1989 and 1999 were reviewed. Information on signalment, onset, progression and duration of clinical signs, diagnostic testing, sites of dorsal laminectomy, postoperative complications, length of hospitalization, and the ambulatory status on discharge was recorded. A minimum follow-up of 7 months was required for inclusion in the study. Neurologic status was graded (0 to 5) preoperatively, 2 days after surgery, and at the time of the study (final score). Improvement or worsening of the neurologic status was assessed by comparison of different scores for each dog. Additional follow-up information was obtained by means of a detailed telephone questionnaire directed at both the owner and referring veterinarian. RESULTS: Mean duration of clinical signs before surgery was 4.9 months. At admission, 15 dogs were ambulatory and 5 were nonambulatory. Neurologic status worsened in 70% of dogs 2 days after surgery but improved in all but 1 dog over the long term. Mean time to optimal recovery was 3.6 months. Long-term follow-up ranged from 7 months to 9 years (mean +/- SD, 3.2 +/- 2.4 years). Four dogs had confirmed recurrence; 2 other dogs may have had recurrence. CONCLUSIONS: Dorsal cervical laminectomy is an effective treatment for CCSM in those dogs with dorsal compression or multiple sites of involvement. CLINICAL RELEVANCE: Although most dogs' neurologic status transiently worsened after surgery, long-term outcome and recurrence rates were comparable to those seen with other surgical techniques for CCSM.     

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.