Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice

We previously induced protective immune response by oral immunization with yeast expressing the ApxIIA antigen. The ApxI antigen is also an important factor in the protection against Actinobacillus pleuropneumoniae serotype 5 infection; therefore, the protective immunity in mice following oral immunization with Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) was compared with that in two control groups (group A and B). The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p < 0.01). The highest systemic (IgG) and local (IgA) humoral immune responses to ApxIA and ApxIIA were detected in group E after the third immunization (p < 0.05). The levels of IL-1β and IL-6 after challenge with an A. pleuropneumoniae field isolate did not change significantly in the vaccinated groups. The level of TNF-α increased in a time-dependent manner in group E but was not significantly different after the challenge. After the challenge, the mice in group E had a significantly lower infectious burden and a higher level of protection than the mice in the other groups (p < 0.05). The survival rate in each group was closely correlated to the immune response and histopathological observations in the lung following the challenge. These results suggested that immunity to the ApxIA antigen is required for optimal protection.     

Bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep

Objective To study the clinical signs following bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep.
Design A clinical and pathological study.
Procedure Twenty Poll Dorset sheep were inoculated with bluetongue virus serotypes 1 or 3, each inoculum having a different passage history. The sheep were examined daily and their clinical appearance and rectal temperatures recorded. Heparinised and non-heparinised blood samples were taken at intervals for virological and serological study. Gross pathological findings were recorded for several sheep at necropsy and tissue samples were collected from three sheep for virological studies.
Results All inoculated sheep developed clinical disease. The clinical signs and gross pathological changes varied considerably but were consistent with damage to the vascular endothelial system. There was a decline in the titres of infectious bluetongue virus and of antigen in tissues collected between 7 and 12 days after infection.
Conclusions The severity of disease was related to the speed of onset and duration of pyrexia and not the development or titre of viraemia. Generally, those animals with sensitive mouths, depression, coronitis, recumbency and reluctance to move were the most debilitated. Whole blood was the most reliable source of infectious virus from acutely and chronically infected and convalescent animals. However, tissue samples particularly spleen, collected from dead or killed animals suffering from either peracute or acute forms of disease were most appropriate for the rapid confirmation of a clinical diagnosis.     

Unguided bronchoalveolar lavage techniques and residual effects in dogs

Objectives To investigate the use of unguided bronchoalveolar lavage techniques in dogs without fibreoptic bronchoscopy, using an adapted single vascular catheter and a double-lumen catheter made from two single vascular catheters.
Animals Sixty-nine dogs were examined with the single-catheter technique and 110 dogs with the double-catheter technique.
Design A prospective study.
Procedure Sixty-nine and 220 samples, collected with the single catheter and the double catheter respectively, were examined cytologically Lungs of 69 dogs were examined grossly and histologically. Radiographic examination was performed on 11 dogs.
Results The double-catheter technique produced samples with significantly higher cellularity (P < 0.01) and fewer red blood cells (P < 0.01) than the single-catheter technique. Repeat samples collected with a double catheter were not significantly different (P > 0.01) in any value. A reference range for nucleated cell counts of 62 to 1210 – 10⁶/L was calculated from 57 clinically and histologically normal dogs. The major residual effects of the technique were localised pulmonary oedema, and alveolar distension with collapse and congestion of distant parenchyma. Thoracic radiographs revealed increased lung opacity for at least up to 7 h after the procedure.
Conclusions The cellularity of the bronchoalveolar lavage fluid obtained was adequate and sufficient fluid was retrieved when the single catheter was located in a proper position. However, the double catheter obtained better samples more quickly and easily, with less damage to the respiratory tract.     

Subject-based problem-based learning in the veterinary science course at the University of Queensland

Objective
To assess students' perceptions of subject-based problem-based learning in the fourth year of the veterinary science course at the University of Queensland.
Design
A questionnaire-based study.
Procedure
Subject-based problem-based learning was introduced into parts of two fourth year subjects in a 5-year veterinary science course. The problem-based learning exercise used modified clinical cases and was computer-assisted. Students worked in groups of two to four, and small group discussion sessions were tutorless. Lectures were replaced by large group discussion and feedback sessions, led by the teacher, with approximately 85 students.
Results
There was a significant increase in the percentage of students who strongly agreed that they had better understanding of the subject, and had learned to apply principles from this class in new situations. The only consistent criticism by students was directed at the extra time required compared to traditional lecture-based subjects.
Conclusion
Students' perceptions of the learning outcome were very favourable for problem-based learning when compared to the lecture-based subject.     

Long- and short-term flooding effects on survival and sink-source relationships of swamp-adapted tree species

About 95% of swamp tupelo (Nyssa sylvatica var. biflora (Walt.) Sarg.) and sweetgum (Liquidambar styraciflua L.) seedlings survived continuous root flooding for more than two years, whereas none of the swamp chestnut oak (Quercus michauxii Nutt.) and cherrybark oak (Q. falcata var. pagodifolia Ell.) seedlings survived one year of flooding. Death of oak seedlings occurred in phases associated with periods of major vegetative growth, e.g., after bud burst in spring, after summer stem elongation, and during the winter deciduous stage, suggesting that stored reserves and sources were inadequate to maintain the seedlings when vegetative sinks were forming. Additional evidence that flooding induced a source deficiency in oak was that leaves of flooded oak were 65 to 75% smaller than leaves of nonflooded oak. Flooded swamp tupelo seedlings had a normal leaf size and patchy stomatal opening compared with nonflooded seedlings. Flooding caused increases in alcohol dehydrogenase (ADH) specific activity in taproot cambial tissues and increases in starch concentrations of swamp tupelo seedlings that were reversed when seedlings were removed from flooding. Flooding had little effect on soluble sugar concentrations in swamp tupelo or sweetgum. In the long-term flood-dry-flood treatment, in which all species had survivors, upper canopy leaf photosynthetic rates were higher in all species during the dry period than in nonflooded controls, whereas their starch and soluble sugars concentrations were similar to those of nonflooded controls. Based on seedling survival and the sink-source relationships, the order of flood tolerance was: swamp tupelo > sweetgum > swamp chestnut oak > cherrybark oak.     

Characterization of proanthocyanidin in hot water extract isolated from <Emphasis Type="Italic">Pinus radiata</Emphasis> bark

Proanthocyanidins (PAs) in the hot water extract prepared from Pinus radiata bark were isolated by a Sephadex LH-20 chromatography. The PAs were characterized using UV/Visible, FT-IR and ¹³C nuclear magnetic resonance (NMR) spectroscopic techniques, matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS), and gel permeation chromatography (GPC) analysis. Bate-Smith acidic alcoholysis and FT-IR spectrum of the PAs indicated the predominant presence of the procyanidin (PC) structure. The ¹³C NMR spectrum revealed that the PAs comprise PC (94%) together with a small amount of prodelphinidin (PD, 6%), and also that they contain the higher trans configuration rather than the cis configuration of C2–C3. The MALDI-TOF MS results obviously proved the presence of PD units, but the maximum degree of polymerization (DP) was limited to octamer. The average molar mass (Mw, Mn), polydispersity (Mw/Mn), and DP of the pine bark PAs were 3,800 (Mw), 1,200 (Mn), 3.2, and 13, respectively.     

Determination of reference intervals for metabolic profile of Hanwoo cows at early,middle and late gestation periods

Background

Metabolic profile was initially designed as a presymptomatic diagnostic aid based on statistical analyses of blood metabolites to provide an early warning of certain types of metabolic disorder. However, there is little metabolic profile data available about Korean Hanwoo cows. Therefore, this study aimed to determine the reference intervals of metabolic profile for Korean Hanwoo cows.

Methods

Healthy animals (2,205) were selected and divided into early (day 1 to 95), middle (day 96 to 190) and late (day 191 to 285) period according to their gestating period. Metabolic profile including total protein (TP), albumin (Alb), urea (UREA), glucose (Glu), total cholesterol (T-Cho), long-chain fatty acid (LCFA), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), creatinine (Crea), calcium (Ca), inorganic phosphorous (iP) and magnesium (Mg) were analyzed using a TBA-40FR automatic biochemical analyzer. The data of Korean Hanwoo cows were then compared to those of the Japanese Wagyu cows.

Results

Most of the data of the Korean Hanwoo cows were relatively higher than those of Japanese Wagyu cows, with the exception of Glu and GGT. This may indicate that the nutritional level of feed for the Korean Hanwoo cows was higher than that of the Japanese Wagyu cows because of the different feeding system. In particular, relatively higher levels of UREA and LCFA were observed in the Korean Hanwoo cows, and this may also contribute to the low reproduction efficiency.

Conclusions

These findings may provide some theoretical basis for understanding the reproductive and feeding situation of Korean Hanwoo cows.     

Antioxidant and antimelanogenic activities of polyamine conjugates from corn bran and related hydroxycinnamic acids

The antioxidant activity of three major polyamine conjugates, N,N'-dicoumaroyl-putrescine (DCP), N-p-coumaroyl-N'-feruloylputrescine (CFP), and N,N'-diferuloyl-putrescine (DFP) isolated from corn bran, and their related hydroxycinnamic acids, p-coumaric acid and ferulic acid, were evaluated by three antioxidant in vitro assay systems, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide and hydroxyl radicals generated by enzymatic and nonenzymatic reactions. Additionally, five phenolic compounds were evaluated for melanogenesis inhibitory activity using mushroom tyrosinase and B16 melanoma cells. Most of the phenolic compounds significantly scavenged DPPH, superoxide, and hydroxyl radicals in a dose-dependent manner. Particularly, DFP showed potent DPPH (IC50 = 38.46 microM) and superoxide (IC50 = 291.62 microM) radical scavenging activities, while DCP exhibited the strongest hydroxyl radical scavenging activity (IC50 = 120.55 microM). CFP also exerted moderate DPPH, superoxide, and hydroxyl radical scavenging activities. Meanwhile, DCP (IC50 = 181.73 microM) showed potent tyrosinase inhibitory activity toward l-tyrosine as the substrate, whereas DFP (IC50 = 733.64 microM) significantly inhibited melanin synthesis in B16 melanoma cells. These current results indicate that these three polyamine conjugates from corn bran may be useful potential sources of natural antioxidants and skin-whitening agents.     

Protective effect of heat-processed American ginseng against diabetic renal damage in rats

We investigated the effects of American ginseng (AG) and heat-processed American ginseng (H-AG) on diabetic renal damage using streptozotocin (STZ)-induced diabetic rats in this study. The diabetic rats showed a loss of body weight gain, and increases in kidney weight, food intake, water intake, and urine volume, whereas the oral administration of H-AG at a dose of 100 mg/kg of body weight per day for 20 days attenuated these diabetes-induced physiological abnormalities. Among the renal function parameters, the elevated urinary protein levels in diabetic control rats were significantly decreased by the AG or H-AG administrations, and the decreased creatinine clearance level was significantly increased in H-AG-administered rats. In addition, the markedly high serum levels of glucose and glycosylated protein in diabetic control rats were significantly decreased by the administration of H-AG, implying that H-AG might prevent the pathogenesis of diabetic complications caused by impaired glucose metabolism and glycosylation of serum proteins. Although no significant ameliorations were shown in overexpressed protein expressions related to diabetic oxidative stress by the AG or H-AG administrations, the accumulation of N (epsilon)-(carboxymethyl)lysine and receptors for advanced glycation endproduct (AGE) expressions were significantly reduced by the administration of H-AG. On the basis of these results, we found that AG and H-AG inhibit AGE accumulation in diabetic rat kidney by their hypoglycemic and renal function ameliorating effects, and this effect was stronger in the H-AG-administered group than in the AG-administered group. These findings indicate that H-AG may have beneficial effect on pathological conditions associated with diabetic nephropathy.     

Characterization of aerosols containing microcystin

Toxic blooms of cyanobacteria are ubiquitous in both freshwater and brackish water sources throughout the world. One class of cyanobacterial toxins, called microcystins, is cyclic peptides. In addition to ingestion and dermal, inhalation is a likely route of human exposure. A significant increase in reporting of minor symptoms, particularly respiratory symptoms was associated with exposure to higher levels of cyanobacteria during recreational activities. Algae cells, bacteria, and waterborne toxins can be aerosolized by a bubble-bursting process with a wind-driven white-capped wave mechanism. The purposes of this study were to: evaluate sampling and analysis techniques for microcystin aerosol, produce aerosol droplets containing microcystin in the laboratory, and deploy the sampling instruments in field studies. A high-volume impactor and an IOM filter sampler were tried first in the laboratory to collect droplets containing microcystins. Samples were extracted and analyzed for microcystin using an ELISA method. The laboratory study showed that cyanotoxins in water could be transferred to air via a bubble-bursting process. The droplets containing microcystins showed a bimodal size distribution with the mass median aerodynamic diameter (MMAD) of 1.4 and 27.8 mum. The sampling and analysis methods were successfully used in a pilot field study to measure microcystin aerosol in situ.