Molecular characterisation of a field strain of bubaline herpesvirus isolated from buffaloes (Bubalus bubalis) after pharmacological reactivation

Two healthy buffaloes (Bubalus bubalis) in a herd which had not been vaccinated against infectious bovine rhinotracheitis (IBR), were selected for their seropositivity for anti-bovine herpesvirus type 1 (BoHV-1) glycoprotein E antibodies, and injected intramuscularly daily with dexamethasone for five consecutive days (day 1 to day 5) to reactivate any latent herpesvirus. Blood samples and nasal and vaginal swabs were collected daily from day 5 to day 15 from each buffalo for virological examination. All the vaginal swabs and blood samples were negative, but 13 of the 22 nasal swabs were positive; a cytopathic effect was observed in primary cultures of bovine fetal lung cells, and the viral isolates were identified as a herpesvirus by PCR. The viral strains were characterised by the sequence analysis of the genes coding for glycoproteins D and B, and the gene sequences were then used for phylogenetic analysis. The isolates from both buffaloes appeared identical at the level of the two genes, and were more closely related to bovine herpesvirus type 5 than to BoHV-1.     

TSL-1 antigens of Trichinella: an overview of their potential role in parasite invasion, survival and serodiagnosis of trichinellosis

The majority of studies on the immunobiology of Trichinella species have centred on the larval muscular phase (L1) with a view to identifying immunodominant antigens located on the surface of the cuticle and in the larval secretions; the nucleus of the parasite-host interaction. These antigens have been classified as eight groups (TSL-1-TSL-8), of which those belonging to the group TSL-1 have been most intensely studied. The principal constituents are glycoproteins, glycan carriers that contain a unusual sugar, the tyvelose (3,6-dideoxy-d-arabinohexose). Studies aimed at improving serodiagnostic techniques to detect trichinellosis indicate that these antigens are ideal candidates. They are capable of inducing a strong humoral response involving the generation of specific antibodies against beta-tyvelose, a sugar that seems to be exclusive to the Trichuroidea. Furthermore, these glycoproteins appear to fulfil an important function in the development and maintenance of the parasite in the muscular niche, and they appear to be fundamental for the invasion of the intestinal epithelium. It has also been demonstrated that specific monoclonal antibodies against tyvelose can mediate a degree of immunoprotection in the rat through the phenomenon known as rapid expulsion.     

Phenylphenalenone type compounds from the leaf fibers of abaca (Musa textilis)

A series of phenylphenalenone type compounds, known to play a role as phytoalexins in plants of the Musaceae family, have been identified for the first time in the leaf fibers of abaca (Musa textilis). Among the phenylphenalenone type compounds identified, the structure of a novel compound, (1R)-2,3-dihydro-4,9-dihydroxy-8-methoxy-1-phenylphenalene, has also been described in abaca fibers. Its structure was elucidated by analysis of one- and two-dimensional NMR (correlation spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond correlation) spectroscopic data.     

Comparative spatial analysis of the sooty blotch/flyspeck disease complex,bull’s eye and bitter rots of apples

The incidence of sooty blotch/flyspeck (SBFS) and bitter and bull’s eye rots were assessed in a Fuji apple orchard during two seasons. Using a regular sampling design, 252 trees were selected and 20 fruits per tree were sampled at harvest and scored for disease incidence. For bitter and bull’s eye rots, additional assessments were made on all symptomless fruit after a 30‐day period of storage. Randomness in the spatial pattern was assessed using beta‐binomial analysis of incidence data for three sampling scales (one, three or six adjacent trees as sampling units) and using Spatial Analysis by Distance Indices (sadie ) for disease counts for the 3‐tree sampling scale. sadie was also used for testing spatial associations between a pair of diseases, between years for the same disease or between rotted and latently infected fruit. Using a toroidal‐shifts procedure, 360 maps of disease counts were created based on the observed data, which were further analysed using sadie . Most datasets showed an aggregated spatial pattern, which was more consistent for the two fruit rots than SBFS, which showed distinct patterns depending on the year or method of analysis. The two fruit rots were spatially associated in most situations but SBFS and bull’s eye rot were dissociated in one season. Results from virtual orchards showed that the patterns observed in the original maps may accurately represent those in similar apple‐growing areas. Hypotheses regarding aspects of ecology and epidemiology of pathogens studied and potential efficacy of control measures in the region are discussed.     

Interleukin-6 and insulin incrase and nitric oxide and adiponectin decrease in blind dogs with pituitary-dependent hyperadrenocorticism

In this study, two populations of dogs with pituitary dependent hypercortisolism (PDH) were compared over a 2-year period. One group had normal vision (Group A, n=27) and one group was blind (Group B, n=20). Group B was characterised by the rapid appearance of the clinical signs of PDH that precede blindness. We found increases in pre-adrenocorticotropic hormone cortisol (P=0.002), IL-6 (P=0.0001), insulin, and insulin sensitivity (detected with the Homeostatic Model Assessment, P<0.0001) in Group B but not in Group A. The nitric oxide (NO) and the total adiponectin concentrations decreased (P=0.0001 and P=0.02, respectively) in Group B versus Group A. The IL-6 and insulin concentrations and the HOMA-A index were positively correlated with the cortisol concentration and were negatively correlated with the NO concentration. With the exception of adiponectin, the other variables were associated with blindness. We concluded that blindness in PDH is a haemodynamic event associated with metabolic changes, with the increase in the IL-6 concentration and the decrease in the NO concentration affecting the retinal vasculature and producing a high risk of vision loss.     

Development and characterization of mouse monoclonal antibodies reactive with chicken CD83

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.     

Influence of diet on calcium metabolism, tissue calcification and urinary sludge in rabbits (Oryctolagus cuniculus)

Rabbits absorb more calcium (Ca) from their diet than they require, and excrete surplus via urine, which therefore contains a typical 'sludge'. This makes rabbits susceptible to Ca-containing uroliths. But given the Ca content of diets of free-ranging specimens, and the limited reports of urinary sludge and Ca contents in free-ranging lagomorphs, we can suspect that rabbits are naturally adapted to high urinary Ca loads. We fed four groups of New Zealand hybrid rabbits [n = 28, age at start 5-6 weeks) pelleted diets consisting of lucerne hay only (L, Ca 2.32% dry matter (DM)], lucerne:oats 1:1 (LG, Ca 1.36%), grass hay only (G, Ca 1.04%), or grass:oats 1:1 (GG, 0.83%) for 25 weeks, with water available ad libitum. Diets were not supplemented with Ca, phosphorus, or vitamin D. Rabbits on diets LG and GG had lower food and water intakes, lower faeces and urine output, grew faster and had higher body mass at slaughter (mainly attributable to adipose tissue). Apparent Ca digestibility decreased in the order L-LG-G/GG. Rabbits on L had larger and heavier kidneys, more urinary sediment at sonography, and a higher urinary Ca content than the other groups. No animal showed signs of urolithiasis/calcinosis at X-ray, sonography, or gross pathology. Kidney/aorta histology only sporadically indicated Ca deposits, with no systematic difference between groups. Under the conditions of the experiment, dietary Ca loads in legume hay do not appear problematic for rabbits, and other factors, such as water supply and level of activity may be important contributors to urolithiasis development in veterinary patients. However, due to the lower Ca content of grass hay, the significantly lower degree of urinary sludge formation, and the significantly higher water intake related with grass hay feeding, grass hay-dominated diets are to be recommended for rabbits in which urolithiasis prevention is an issue.     

Field evaluation of a new commercially available ELISA based on a recombinant antigen for diagnosing Chlamydophila abortus (Chlamydia psittaci serotype 1) infection

A new commercially available ELISA (ELISAr-Chlamydia) for detecting antibodies against Chlamydophila abortus has been evaluated using sheep field serum samples. The ELISA is based on a recombinant antigen which expresses part of a protein from the 80-90kDa family that is specific to C. abortus. Sera (105) from six flocks with confirmed ovine chlamydial abortion (OEA) outbreaks were used in this study, as well as sera (258) from 18 flocks which had suffered no OEA in the last lambing. The ELISAr-Chlamydia was compared with the complement fixation test (CFT) and with an ELISA using purified C. abortus elementary bodies (ELISA-EB), employing as reference technique a comparative microimmunofluorescence test that differentiates C. abortus infection from Chlamydophila pecorum infection. The results showed that the sensitivity of ELISAr-Chlamydia was 90.9% with a specificity of 85.9%, the sensitivity of CFT was 71.0% with a specificity of 83.6%, while the sensitivity of ELISA-EB was 95.2% and the specificity was 54.2%. Furthermore, ELISAr-Chlamydia was the test with fewer false positives resulting from positive reactivity to C. pecorum, although 15% of the sera positive for C. pecorum but negative for C. abortus antibodies reacted positively. This study demonstrated with field material that ELISAr-Chlamydia provides the most balanced results between sensitivity and specificity, especially in flocks with no clinical OEA but reactivity to C. abortus.     

A genetic map of an interspecific diploid pseudo testcross population of coffee

Coffee is globally one of the most important export crops and is a prominent part of the economy in more than 50 countries in Latin America, Africa and Asia. In Colombia, it has been the leading export commodity for more than a century. However, genetic research on coffee has been rather sparse and mainly focused on the two major cultivated species, Coffea arabica L. and C. canephora P., leaving unexplored the genetic potential in other species. In this study, an interspecific mapping population consisting of 101 F₁ hybrid plants from a cross between the diploid species C. liberica and C. eugenioides was evaluated for genetic segregation at 618 molecular marker loci. Of these, 168 SSRs and two ESTs exhibited polymorphic patterns that allowed segregation analysis and genetic linkage estimations. A genetic map consisting of 146 co-dominant loci and 11 predicted linkage groups was constructed using the mapping software JoinMap 3.0. The conjoined maternal/paternal map length is 798.68 cM, has an average saturation density of 6.01 cM/interval, and covers an estimated 66–86 % of the diploid coffee genome. Approximately 24 % of loci had null alleles, and 23.5 % exhibited segregation distortion. Knowledge derived from this study has important applications for quantitative trait locus analysis and marker-assisted selection in Colombian coffee breeding programs.