Construction of a microarray specific to the chicken immune system: profiling gene expression in B cells after lipopolysaccharide stimulation

The objective of this study was to profile gene expression in cells of the chicken immune system. A low-density immune-specific microarray was constructed that contained genes with known functions in the chicken immune system, in addition to chicken-expressed sequence tags (ESTs) homologous with mammalian immune system genes, which were systematically characterized by bioinformatic analyses. Genes and ESTs that met the annotation criteria were amplified and placed on a microarray. The microarray contained 84 immune system gene elements. As a means of calibration, the microarray was then used to examine gene expression in chicken B cells after lipopolysaccharide stimulation. Differential gene expression was observed at 6, 12, and 24 h but not at 48 h after stimulation. The results were validated by semiquantitative polymerase chain reaction. The microarray showed a high degree of reproducibility, as demonstrated by intra- and interassay correlation coefficients of 0.97 and 0.95, respectively. Thus, the low-density microarray developed in this study may be used as a tool for monitoring gene expression in the chicken immune system.     

The effects of cold stress on neonatal calves. II. Absorption of colostral immunoglobulins.

Newborn calves were subjected to cold stress and made hypothermic by immersion in water at 15 to 17 degrees C. Cold stress delayed the onset and significantly decreased (P less than 0.05) the rate of absorption of immunoglobulins (IgM, IgG1 and IgG2) up to 15 hours after first feeding of pooled colostrum. However, the net absorption of colostral immunoglobulins was not affected. The possible deleterious effect of cold stress on absorption of colostral immunoglobulins by newborn calves under range conditions is discussed.     

Genetic typing of classical swine fever virus

Three regions of the classical swine fever virus (CSFV) genome that have been widely sequenced were compared with respect to their ability to discriminate between isolates and to segregate viruses into genetic groups. Sequence data-sets were assembled for 55 CSFVs comprising 150 nucleotides of the 5' non-translated region, 190 nucleotides of the E2 envelope glycoprotein gene and 409 nucleotides of the NS5B polymerase gene. Phylogenetic analysis of each data-set revealed similar groups and subgroups. For closely related viruses, the more variable or larger data-sets gave better discrimination, and the most reliable classification was obtained with sequence data from the NS5B region. No evidence was found for intertypic recombination between CSFVs. A larger data-set was also analysed comprising 190 nucleotides of E2 sequence from 100 CSFVs from different parts of the world, in order to assess the extent and global distribution of CSFV diversity. Additional groups of CSFV are evident from Asia and the nomenclature of Lowings et al. (1996) [Lowings, P., Ibata, G., Needham, J., Paton, D., 1996. J. Gen. Virol. 77, 1311-1321] needs to be updated to accommodate these. A tentative assignment, adapting rather than overturning the previous nomenclature divides CSF viruses into three groups with three or four subgroups: 1.1, 1.2, 1.3; 2.1, 2.2, 2.3; 3.1, 3.2, 3.3, 3.4. The expanding data-base of CSFV sequences should improve the prospects of disease tracing in the future, and provide a basis for a standardised approach to ensure that results from different laboratories are comparable.     

Field efficacy of a combined use of Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae vaccines in growing pigs

The effectiveness of simultaneous administration of commercial Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae vaccines was tested in an indoor commercial piggery which had experienced continuing respiratory-disease problems confirmed as due to both of these pathogens. Piglets were randomly assigned in equal numbers to vaccination and control groups, and each vaccine was administered at a separate site to assigned piglets at two and four weeks of age. Live weight of vaccinates immediately prior to slaughter was 2.49 kg higher (p = 0.04) than for controls at equal mean slaughter age of 132 days. Average daily gain (ADG) from 16 weeks to slaughter of vaccinates was also significantly higher (33 g/day) than in controls (p = 0.05). Daily gain was not significantly different in younger age groups. Active enzootic pneumonia lesions were more likely in control than in vaccinated pigs. There were no significant differences between vaccination groups with regard to severity of pleurisy or presence of pleuropneumonia lesions at slaughter. Log-linear modelling was used to test the statistical association between vaccination, enzootic pneumonia lesions, pleurisy lesions and pleuropneumonia lesions. It showed a reduction in the severity of enzootic pneumonia lesions for vaccinated pigs, and the presence of pleuropneumonia lesions increased the likelihood of pleurisy lesions. No other association was significant, and no evidence of synergy between the vaccines in influencing lesion severity for pleuropneumonia was detected (within the limitations set by the trial design).     

Effects of supplemental protein type on productivity of primiparous beef cows

Effects of supplemental degradable (DIP) and undegradable (UIP) intake protein on forage intake, BW change, body condition score (BCS), postpartum interval to first estrus, conception rate, milk production and composition, serum metabolites and metabolic hormones, and calf gain were determined using 36 primiparous Gelbvieh x Angus rotationally crossed beef cows. On d 3 postpartum, cows (average initial BW = 495 +/- 10 kg and BCS = 5.5 +/- 0.1) were randomly assigned to one of three dietary supplements (12 cows/treatment). Date of parturition was evenly distributed across treatment (average span of calving date among treatments = 2.4 +/- 2.5 d). Individually fed (d 3 through 120 postpartum) dietary supplements were 0.82 kg of corn and 0.23 kg of soybean meal per day (DIP), the DIP + 0.12 kg of blood meal and 0.13 kg of corn gluten meal per day (DIP + UIP), and 0.82 kg of corn, 0.07 kg of blood meal, and 0.08 kg of corn gluten meal per day in an isonitrogenous replacement of soybean meal (UIP IsoN). Cows had ad libitum access to native grass hay (8.5% CP) and trace-mineralized salt. Total OM intake was greater (P = 0.06) for DIP + UIP than UIP IsoN cows. At 30 d postpartum, DIP + UIP cows produced more milk than UIP IsoN, with DIP being intermediate; however, at 60 d postpartum, DIP + UIP and DIP cows were not different, but both had greater milk production than UIP IsoN (treatment x day interaction; P = 0.08). A treatment x day interaction (P = 0.06) for BCS resulted from DIP + UIP cows having the greatest BCS at 60, 90, and 120 d d postpartum and DIP having greater BCS than UIP IsoN cows only on d 60 postpartum. Serum insulin concentrations were highest (treatment x day interaction; P = 0.09) for DIP + UIP cows at 30 d postpartum but did not differ among treatment thereafter. Serum insulin-like growth factor-binding protein (IGFBP)-2 (34 kDa) and -3 (40 and 44 kDa) were greatest (P < 0.0003) for DIP cows. Serum urea-N concentrations were greater (P < 0.01) in DIP + UIP cows than in either DIP or UIP IsoN cows. However, postpartum interval to first estrus, conception rate, and calf weaning weights were unaffected (P = 0.35, 0.42, and 0.64, respectively) by treatment. Although UIP in addition to or in replacement of DIP affected milk production and blood metabolites, the productivity of these primiparous beef cows was not altered. Thus, the type of supplemental protein does not seem to influence productivity of primiparous beef cows in production systems with conditions similar to our experimental conditions.     

Intergeneric hybrids and an amphiploid between Elymus canadensis and Psathyrostachys juncea

Summary Fifty four hybrid plants between Elymus canadensis and Psathyrostachys juncea were obtained by handpollination and embryo culture. The average cross compatibility between both species was 31.2 percent. One amphiploid plant was induced by colchicine treatment. The hybrid and amphiploid plants resembled P. juncea in appearance but showed a higher plant height and dry matter yield than the parents. The hybrids showed extremely low pollen stainability and were completely sterile. With the exception of one plant (2n=3x+1=22), all hybrid plants were allotriploids (SHN, 2n=3x=21). The amphiploid plant (SSHHNN, 2n=6x=42) showed 58.9% pollen stainability and 11.6% seed fertility.Mean chromosome associations of the hybrids and amphiploid at metaphase I were 0.02IV+0.06III+2.03II+16.91I and 0.07III+18.00II+5.85I, respectively. Lagging chromosomes, chromosome bridges, abnormal cytokinesis, and micronuclei were occasionally observed at the anaphase, telophase, or tetrad stage.     

In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

Extramedullary plasmacytoma and immunoglobulin-associated amyloidosis in a cat

Extramedullary plasmacytoma with immunoglobulin-associated amyloidosis was diagnosed in a 10-year-old cat. The primary tumor was a large, circumferential mass of the right tarsal region. Metastasis developed in the regional lymph nodes, spleen, and liver. Permanganate-resistant amyloid deposits were associated with plasma cells in the primary and metastatic tumors. Treatment with prednisone and melphalan had little effect on the progression of the disease, and the cat died four weeks later.