Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism

The efficacy of four different commercial live vaccines (vaccines A, B, C, and D) against the infectious laryngotracheitis virus (ILTV) was assessed in specific-pathogen-free (SPF) chickens. SPF chickens were vaccinated intraocularly at 6 wk old with ILTV live vaccines and were challenged intratracheally with the N91B01 strain of virulent Korean ILTV 2 wk after vaccination. The immunity against ILTV live vaccines was assessed by the incidence of latent infection by the challenge virus in the chickens' tracheas and trigeminal ganglia, the reisolation rate of the challenge virus, and the clinical signs in the chickens challenged with the N91B01 strain of ILTV. The latent infection in chickens was assessed by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our data showed that the clinical signs and challenge virus isolation were negative in all chickens receiving four difference commercial ILTV live vaccines. The viral DNA of the vaccine strain, but not that of the challenge virus, was detected in chickens vaccinated with vaccine A by nested PCR-RFLP. The viral DNAs of both the vaccine and challenge strains were detected from chickens vaccinated with vaccines B, C, and D. This study showed that only vaccine A can protect chickens from latent infection with the field virulent ILTV. We speculate that the efficacy of infectious laryngotracheitis live vaccines to protect chickens from latent infection with virulent ILTVs can be assessed by nested PCR-RFLP analysis.     

Intestinal immunomodulation by vitamin A deficiency and lactobacillus-based probiotic in Eimeria acervulina-infected broiler chickens

In a 2 X 2 factorial study, a broiler starter ration was amended for vitamin A (control, C; deficient, A) and probiotic status (-, P) to investigate their modulatory effects onthe host immune system. Birds were inoculated orally with Eimeria acervulina (EA) oocysts, and disease susceptibility was evaluated by assessment of fecal oocyst shedding. Humoral and local cellular mediated immunity were assessed by evaluation of antibody and cytokine (interferon-gamma [IFN-gamma] and interleukin-2 [IL-2]) levels in sera and intestinal secretions on a 3-day interval after inoculation. Fecal oocyst shedding was highest (P < 0.05) in A- birds, followed by AP, C-, and CP birds. Feeding the probiotic reduced shed oocysts by 20% in A fed birds and by 26% in C fed birds. Intestinal IFN-gamma was relatively constant in all treatment groups except for A-, where it declined steadily and was lower (P < 0.05) from day 6 on. Serum IFN-gamma levels fluctuated within each treatment and over time were not revealing. Intestinal IL-2 was highest in CP birds at 3 and 9 days postinfection (DPI) and lowest in A- birds at 3, 9, and 12 DPI (P < 0.05); no difference between treatments was found at 6 DPI (P > 0.05). Eimeria-specific intestinal antibody (Ab) level was constant (P > 0.05) in C- birds but increased with time (P < 0.05) in A-, AP, and CP birds. Serum Ab levels were also constant in A- and CP birds but increased (P < 0.05) in C- and AP birds after 6 DPI. The data demonstrate for the first time a probiotic-enhanced immunity in vitamin A deficient birds. This study is also the first to demonstrate the probiotic effect on local cell-mediated immunity of chickens, best manifested by apparent lower intestinal invasion and development by EA, on the basis of higher IL-2 secretion and lower EA oocyst production.     

Anti-protozoal efficacy of medicinal herb extracts against Toxoplasma gondii and Neospora caninum

The purpose of this study was to determine whether alcohol extracts of herbs (Sophora flavescens Aiton, Sinomenium acutum (Thunb.) Rehder and E.H. Wilson, Pulsatilla koreana (Yabe ex Nakai) Nakai ex T. Mori, Ulmus macrocarpa Hance and Torilis japonica (Houtt.) DC.) from South Korea, possess in vitro anti-protozoal activity against cultures of Toxoplasma gondii and Neospora caninum. These herbs have been used as human anti-parasitics in Asian countries for many years. Alcohol extracts of these herbs were serially diluted to final concentrations ranging from 625 to 19.5 ng/ml in media and added to wells containing either T. gondii or N. caninum tachyzoites in equine dermal (ED) cells. Parasite growth inhibition was measured using 3H-uracil incorporation as compared to untreated controls. T. japonica inhibited T. gondii proliferation by 99.3, 95.5, 73.0 and 54.0% in the range from 156 to 19.5 ng/ml, and S. flavescens inhibited T. gondii proliferation by 98.7, 83.0 and 27.2% in the range from 156 to 39 ng/ml. T. japonica inhibited N. caninum proliferation by 97.8, 97.9, 85.3 and 46.4% in the range from 156 to 19.5 ng/ml. S. flavescens inhibited N. caninum proliferation by 98.6, 97.0, 69.5 and 14.0% in the range from 156 to 19.5 ng/ml. Toxicity to host cells was noted when concentrations of T. japonica and S. flavescens exceeded 625 ng/ml. The herb extracts from S. acutum, Pulsatilla koreana, and U. macrocarpa also showed toxicity at higher levels but did not achieve the same inhibition effects at the lower concentrations against T. gondii and N. caninum as T. japonica and S. flavescens.     

Pax-7 immunoreactivity in the post-natal chicken central nervous system

In this immunocytochemical study on the constitutive expression of Pax-7 protein in the postnatal chicken brain, Pax-7 showed region and cell type specific expression. In the optic tectum, only cells in grey matter showed positive immunoreactivities (IRs), whereas those in the white matters did not show any IRs. In thalamic nuclei and several pontine nuclei, we also localized Pax-7 positive IRs. On the contrary, in the cerebellum, Pax-7 was mainly localized within the Bergmann glia, whereas Purkinje cells did not show any IRs. In double immunolabelling studies, most of the Pax-7 IRs did not originate from neuroglial cells such as oligodendrocytes, microglia or astrocytes, but from neurons, with the exception of Bergmann glia in the cerebellum. The presence of Pax-7 IRs in the adult chicken brain could suggest that Pax-7 might play a role in maintaining normal physiological function in some postnatal chicken brain cells.     

Characterization of an avianpox virus isolated from an Andean condor (Vultur gryphus)

A novel pox virus, condorpox virus (CPV) isolated from the spleen of an Andean condor (Vultur gryphus) by inoculation of chorioallantoic membranes (CAM) of specific pathogen free (SPF) chicken embryos was compared biologically, antigenically and genetically with fowlpox virus (FPV), the type species of the genus Avipoxvirus. Susceptible chickens inoculated with CPV developed only mild localized lesions but were not protected against subsequent challenge with FPV. Based on Western blotting, in addition to the presence of cross-reacting antigens, distinct differences in antigenic profiles of CPV and FPV were observed. Sequence analysis of a 4.5 kb HindIII fragment of CPV genomic DNA revealed the presence of eight co-linear genes corresponding to FPV open reading frame (ORF)193-198, 201 and 203. Interestingly, reticuloendotheliosis virus (REV) sequences present in the genome of all FPV were absent in CPV. Although, the results of a phylogenic analysis suggested that CPV is a member of the genus Avipoxvirus, its unique antigenic, biologic and genetic characteristics distinguish it from FPV to be considered as a new member of this genus.     

Genetic and antigenic characterization of a poxvirus isolate from ostriches

Avian poxvirus was isolated from nodules on the heads and conjunctiva of two 3-to-4-wk-old ostrich chicks. The ostriches from which poxvirus was isolated had been placed on premises where turkeys that had shown evidence of poxvirus infection had been raised earlier. Microscopically, the nodules from the ostriches were composed of proliferating and hypertrophic epithelial cells that formed large fronds. Most of the hypertrophic epithelial cells contained large eosinophilic intracytoplasmic inclusion bodies characteristic of poxvirus. Characterization of the avian poxvirus isolated from the cutaneous lesions in ostriches was based on western blotting of virus antigen, restriction fragment length polymorphism of genomic DNA, pathogenesis, and cross-protection studies in chickens. Antigenic and genetic studies did not reveal any significant difference between the poxvirus isolated from ostriches (PVO) and fowl poxvirus (FPV). Further, susceptible chickens immunized with the PVO were protected when challenged with a virulent strain of FPV. Thus, the poxvirus isolated from ostriches had similar antigenic, genetic, and biological properties to FPV.     

Effect of group size and feeder type on growth performance and feeding patterns in finishing pigs

The effects of four group sizes (2, 4, 8, and 12 pigs per pen) and two single-space feeder types (conventional and electronic feed intake recording equipment [FIRE]) on feed intake, growth performance, and feeding patterns were determined in 208 crossbred finishing pigs (equal numbers of barrows and gilts) between 84.4 (SD = 0.81) to 112.8 (SD = 1.08) kg BW over a 4-wk period. Pigs were given ad libitum access to a corn-soybean meal-based diet (15.9% CP; 0.79% lysine; 3,328 kcal ME/kg). The floor space allowance was 0.9 m2/pig for all treatments. Growth rates were not different for the two feeder types; however, feed intake was lower and gain:feed ratio higher for pigs on the FIRE feeders (P < 0.01). Feed intake, growth rate, and gain:feed ratio were not different (P > 0.05) among the group sizes. Number of feeder visits per day decreased and feed intake per visit, feeder occupation time per visit, feed consumption rate, and percentage of time the feeder was occupied increased with group size (P < 0.05). Feed intake per visit had the strongest correlation with daily feed intake (r = 0.54; P < 0.01) and was negatively correlated with gain:feed ratio (r = -0.38; P < 0.01). However, the correlations between growth performance and other feeding pattern traits were relatively weak (r < or = 0.30). As group size increased, diurnal variation in number of feeder visits and feed consumed per hour decreased. There was no difference in time spent sitting and standing between the two feeder types. The proportion of time spent eating was generally lower for the larger groups on both feeders. The proportion of time spent lying was similar across group sizes for pigs on the conventional feeders but was greater for pigs in the larger groups on the FIRE feeders. This study suggests that finishing pigs can maintain feed intake and growth rate by changing feeding behavior as group size increases from 2 to 12.     

The nutritional value of degermed,dehulled corn for pigs and its impact on the gastrointestinal tract and nutrient excretion

Three experiments were designed to assess the feeding value and potential environmental benefits of feeding degermed, dehulled corn, a low fiber by-product originating from the corn dry milling process, to pigs. Twelve 27-kg (SE = 0.8) barrows were used in Exp. 1 to measure the apparent fecal digestibility of DM, GE and N of degermed, dehulled corn compared with corn grain. Two diets were formulated to contain either 96.4% of degermed, dehulled corn or corn grain plus supplemental vitamins and minerals. Digestibilities of DM, GE, and N were greater in degermed, dehulled corn (96.2, 96.0, and 93.6%, respectively) compared with corn grain (89.0, 89.0, and 78.4%, respectively) (P < 0.01). Overall, a 67 and 29% reduction in DM and N excretion, respectively, was observed. In Exp. 2, eight 70-kg (SE =1.8) barrows were surgically fitted with ileal cannulae and fed the same diets as in Exp. 1, to measure the ileal digestibility of nutrients in degermed, dehulled corn. Ileal digestibility of DM, energy, and N was 13, 15, and 7% greater in degermed, dehulled corn (P < 0.05). Apparent ileal digestibility coefficients of leucine, methionine, and phenylalanine were greater in degermed, dehulled corn compared with corn grain (P < 0.05) while a trend for a lower tryptophan digestibility in degermed, dehulled corn was observed (P = 0.067). In Experiment 3, 96 nursery pigs with an initial average BW of 8.8 kg (SE = 0.08), fed a starter diet formulated with degermed, dehulled corn or corn grain as the major grain source, were used in a 28-d growth performance study. At the end of the study, 24 pigs (1 pig per pen) were sacrificed and gastrointestinal tract measurements were taken. Daily growth rates of pigs were the same between diets (0.64 kg/d). A trend for reduced feed intake (P = 0.073) in pigs fed degermed, dehulled corn led to a 4% improvement in gain to feed (P < 0.05). Feeding degermed, dehulled corn had no effect on gut fill, gastrointestinal tract weight, or liver weight (P > 0.05). Ileal villus lengths and crypt depths were not affected by feeding degermed, dehulled corn although ileal villus widths were greater in pigs fed corn grain. Results from these trials suggest that corn processed to remove poorly digestible fiber fractions provides more digestible nutrients than corn grain. As a result, degermed, dehulled corn reduces fecal and N excretion, thus providing a means to reduce nutrient excretion.     

Low expression of cyclin D2 in G2/M-arrested and transformed proliferating Balb/3T3 cells

At present, the effect of cyclin D2 implicated in cell cycle regulation, differentiation, and oncogenic transformation is not fully confirmed. To better elucidate the role of cyclin D2 in controling the cell proliferation, cyclin D2 expression level was determined at the early initiation and promotion stages during the in vitro two-stage transformation process of Balb/3T3 A31 cells. N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced G2/M-arrested cells expressed low level of cyclin D2 mRNA, while the contact-inhibited nonproliferating cells expressed high level of cyclin D2 mRNA. In the transformed proliferating cells at the promotion stage, cyclin D2 mRNA was not expressed. These data suggest that cyclin D2 expression may be associated with the type of growth arrest and nonproliferating state, but not with the cell proliferation and transformation.