Immune responses to a Staphylococcus aureus GapC/B chimera and its potential use as a component of a vaccine for S. aureus mastitis

Bovine mastitis caused by strains of S. aureus is the most economically important disease affecting the dairy industry worldwide. Commercially available vaccines show various degrees of success and work in research laboratories with experimental vaccines suggests that in part, the failure of these vaccines lies in the limited antigenic repertoire contained in the vaccine formulations. Since it seems impractical to produce a vaccine containing antigens from all major S. aureus mastitis isolates, we took the approach of using two surface antigens GapB and GapC that appear to be conserved and constructed a GapC/B chimera as the basis for a vaccine. The humoral and cellular immune responses to GapC/B were compared to the responses to the individual proteins, alone or in combination. The GapC/B protein elicited strong humoral and cellular responses in mice as judged by the levels of total IgG, IgG1, IgG2a, and number of IL-4- and IFN-gamma-secreting cells. These results suggest that this chimeric protein could be an attractive target for further vaccine efficacy studies.     

Effect of changes in ionized calcium concentration in arterial blood and metabolic acidosis on the arterial partial pressure of oxygen in dogs

OBJECTIVE: To evaluate the effects of metabolic acidosis and changes in ionized calcium (Ca2+) concentration on PaO2 in dogs. ANIMALS: 33 anesthetized dogs receiving assisted ventilation. PROCEDURE: Normal acid-base status was maintained in 8 dogs (group I), and metabolic acidosis was induced in 25 dogs. For 60 minutes, normocalcemia was maintained in group I and 10 other dogs (group II), and 10 dogs were allowed to become hypercalcemic (group III); hypocalcemia was then induced in groups I and II. Groups II and IV (5 dogs) were treated identically except that, at 90 minutes, the latter underwent parathyroidectomy. At intervals, variables including PaO2, Ca2+ concentration, arterial blood pH (pHa), and systolic blood pressure were assessed. RESULTS: In group II, PaO2 increased from baseline value (96 +/- 2 mm Hg) within 10 minutes (pHa, 7.33 +/- 0.001); at 60 minutes (pHa, 7.21 +/- 0.02), PaO2 was 108 +/- 2 mm Hg. For the same pHa decrease, the PaO2 increase was less in group III. In group I, hypocalcemia caused PaO2 to progressively increase (from 95 +/- 2 mm Hg to 104 +/- 3 mm Hg), which correlated (r = -0.66) significantly with a decrease in systolic blood pressure (from 156 +/- 9 mm Hg to 118 +/- 10 mm Hg). Parathyroidectomy did not alter PaO2 values. CONCLUSIONS AND CLINICAL RELEVANCE: Induction of hypocalcemia and metabolic acidosis each increased PaO2 in anesthetized dogs, whereas acidosis-induced hypercalcemia attenuated that increase. In anesthetized dogs, development of metabolic acidosis or hypocalcemia is likely to affect ventilatory control.     

Correlation of magnetic resonance images with anatomic features of the equine tarsus

OBJECTIVE: To correlate anatomic features of the equine tarsus identified in plastinated sections with images obtained via magnetic resonance imaging (MRI). ANIMALS: 4 horses. PROCEDURE: MRI (1.5-Tesla magnet) of the tarsus was performed on the pelvic limbs of 4 clinically normal horses following euthanasia. After imaging, tarsocrural joint spaces and vasculature were injected with colored latex. Sagittal and transverse sections of the tarsi were plastinated to facilitate interpretation of MR images. RESULTS: Relevant anatomic structures were identified and labeled on the plastinated tissue slices and corresponding MR images. Results indicated high correlations between MRI findings and those of plastinated sections. CONCLUSIONS AND CLINICAL RELEVANCE: The data obtained provided certain reference standards for normal anatomic structure sizes and positions in the equine tarsus. This information may aid future physiologic or clinical studies of this joint.     

The fracture toughness and properties of thermally modified beech and ash at different moisture contents

The fracture toughness of thermally modified beech (Fagus sylvatica L) and ash (Fraxinus excelsior L) wood under Mode I loading was quantified using Compact Tension (CT) specimens, loaded under steady-state crack propagation conditions. The influence of three heat-treatment levels and three moisture contents, as well as two crack propagation systems (RL and TL) was studied. Complete load–displacement records were analysed, and the initial slope, k _init, critical stress intensity factor, K _Ic, and specific fracture energy, G _f, evaluated. In the case of both species, thermal modification was found to be significantly affect the material behaviour; the more severe the thermal treatment, the lower the values of K _Ic and G _f, with less difference being observed between the most severe treatments. Moisture content was also found to influence fracture toughness, but had a much less significant effect than the heat treatment.     

Homegarden agroforestry systems: an intermediary for biodiversity conservation in Bangladesh

Biodiversity conservation is one of the important ecosystem services that has been negatively impacted by anthropogenic activities. Natural forests (NF) harbor some of the highest species diversity around the world. However, deforestation and degradation have resulted in reduced forest land cover and loss of diversity. Homegarden agroforestry (AF) systems have been proven to be an intermediary for biodiversity conservation. In this study, we evaluate the effectiveness of home garden AF practices to conserve tree species diversity in Bangladesh and compare them with tree species diversity in NF. A total of nine locations were selected for this synthesis from published literature which comprised of five AF sites and four NFs. Shannon?CWeiner Diversity Index (H) was similar for home-garden AF (3.50) and NF (2.99), with no statistical difference between them. Based on non-metric multi-dimensional scaling (NMDS) ordination analysis, the AF and NF plots showed distinct separation. However, Bray?CCurtis dissimilarity index ranged from 0.95 to 0.70 indicating nearly no overlap in species composition to significant overlap between AF and NF. Based on our results, we conclude that AF can serve as an important ecological tool in conserving tree species diversity, particularly on landscapes where NF fragments represent only a small fraction of the total land area. Creating and maintaining AF habitats in such human dominated landscapes should be part of the biodiversity conservation strategy.     

Deep planting with shelters improves performance of different stocktype sizes under arid Mediterranean conditions

New Forests - Increasing plant size has been considered a way to improve survival and growth of planted containerized stock under dry conditions. Additionally, deep planting provides advantages to...     

Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass (Dicentrarchus labrax) during the course of an experimental challenge

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain (cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.     

Cysteine proteinases Fas1 and Fas2 are diagnostic markers for Fasciola hepatica infection in alpacas (Lama pacos)

Circulating antibody against Fasciola hepatica antigens was determined by enzyme-linked immunosorbent assay (ELISA) and immunoelectrophoresis in alpacas naturally exposed to F. hepatica. Serological assay parameters were established by using sera from eight infected animals and seven controls with no record of this parasitic infection. Excretory--secretory (ES-) products, Fas1- and Fas2-ELISA were used to survey 307 alpacas from a F. hepatica endemic area in the Peruvian Andes. Seroprevalence of F. hepatica infection varied from 56.7, 64.8 and 66.8% measured by Fas1-, Fas2- and ES-ELISA, respectively. The sensitivity for ES-ELISA was 95%, corresponding Fas1- and Fas2-ELISA sensitivity values were 90 and 95%. In this population, 7% of animals were positive for F. hepatica eggs in faeces, other parasites detected were Trichuris sp. (40%), Nematodirus sp. (34.6%), Lamanema sp. (12.8%) and Eimeria sp. (11.8%). The results show that F. hepatica infected animals elicit circulating antibodies against ES, Fas1 and Fas2. Fas2-ELISA may be proposed as a sensitive assay for the immunodiagnosis of fasciolosis in alpacas.     

Characterization of the functional domain of major surface protein 1a involved in adhesion of the rickettsia Anaplasma marginale to host cells

The major surface protein (MSP) 1a of the genus type species Anaplasma marginale (Rickettsiales: Anaplasmataceae) has been shown to mediate adhesion, infection and transmission of the organism, as well as to contribute to protective immunity in cattle. MSP1a contains a variable number of tandemly repeated peptides in the amino-terminal region, while the remainder of the protein is highly conserved among isolates. The number of repeats varies among geographic isolates of A. marginale but is constant within an isolate and has been used as a stable genetic marker of isolate identity. Because the sequence of the tandem repeats is the most variable part of the protein among isolates, this region of the protein is most likely to be involved in adhesion to host cells, a prerequisite to infection. The purpose of this study was to characterize the organization and function of the MSP1a tandem repeats of A. marginale in adhesion to host cells. We demonstrated by use of recombinant mutant proteins that the tandemly repeated region of MSP1a was necessary and sufficient to mediate adhesion of MSP1a to tick cells and bovine erythrocytes. Synthetic peptides representing the predominant sequences of individual repeats were tested for their adhesive capacity for tick cell extract (TCE). Peptides containing acidic amino acids D or E at position 20 bound to TCE, while peptides with a G as the 20th amino acid were not adhesive to TCE. Antibodies produced in rabbits against a synthetic repeat peptide neutralized A. marginale infection of cultured tick cells, and the neutralization observed was similar to that effected by antibodies produced against the whole MSP1a recombinant protein. Analysis of tandemly repeated MSP1a peptides of several geographic isolates of A. marginale revealed a complex relationship between the msp1alpha genotype and the tick-transmissible phenotype of the isolate and suggested that both the sequence and conformation of the repeated peptides influenced the adhesive properties of MSP1a. These studies demonstrated that the tandemly repeated region of the protein mediates the adhesive function of MSP1a.