The effect of different isolation procedures on canine leucocyte populations and on lectin-induced lymphocyte proliferation

Proliferation assays performed on peripheral blood mononuclear cells (PBMC) are commonly used in experimental and clinical immunology. A prerequisite for an in vitro assay is the ability to obtain relatively pure populations of mononuclear cells from whole blood, as contaminating polymorphonuclear cells may affect the proliferation of lymphocytes. Purification of canine leucocytes from whole blood is associated with difficulties in obtaining pure lymphocytes in high yields. The aim of this study was to optimize the lymphocyte purification from canine whole blood in terms of total cell recovery and purity, while not influencing the proliferation capacity of the isolated cells. To acquire optimal isolation of canine lymphocytes several density gradient media of different densities and osmolalities were examined. For optimal phagocyte removal, pre-treatment of whole blood with carbonyl iron/arabic gum and/or adherence to fibrinogen pre-coated polystyrene tissue flasks were examined. Lectin-induced proliferation was used as measurement of cell activity of the obtained cell fractions after the different separation procedures. Canine blood pre-treated with carbonyl iron/arabic gum followed by density gradient centrifugation with medium 'G' (density: 1.079 g/cm(3), osmolality: 256 mOsm) and adherence to pre-coated polystyrene tissue flask obtained the best PBMC cultures with a median lymphocyte purity of 88% and a median yield of recovered lymphocytes of 54%. This culture also resulted in the highest proliferation and subsequently the highest stimulation index upon lectin stimulation.     

Research note: Evaluation of the reliability of local farmer knowledge in Namaqualand,South Africa

Abstract

Local farmer knowledge of species distribution patterns and plant palatability in Namaqualand, South Africa, was assessed using Participatory Rural Appraisal techniques. These findings were compared with the results obtained using phytosociological techniques (species distribution), and with palatability indices published in the literature. It was found that farmers were knowledgeable of species distribution close to the settlement, but less so further away from the settlement. Farmer plant palatability rankings did not correlate well with the palatability indices found in the published literature, which suggests that palatability in semi‐arid regions are locality specific.     

Retrieval of a patent ductus arteriosus coil following embolisation to the right subclavian artery

A two-month-old female Tibetan terrier was re-presented with an incomplete closure of the patent ductus arteriosus. Following a second attempt to close the shunt by coil embolisation, arterial embolisation occurred. The successful removal of the embolised coil is described.     

Requirement of the hen for sulphur‐containing amino acids

1. The sulphur amino acid requirements of White Leghorn‐type laying hens were determined in two 52‐week experiments using diets based on maize and soyabean meal providing from 5.0 to 6.5 g sulphur amino acids/kg, 12.35 MJ ME/kg and 8.6 to 8.8 g lysine/kg. There were six replicates of 15 individually‐caged hens each per treatment.

2. A requirement of approximately 775 to 800 mg sulphur amino acids /hen d, of which about 390 to 440 mg was methionine, was found for a maximum of 80 to 83 eggs/100 hen d. There were indications that the requirement for maximum egg production was less than that for maximum efficiency of food utilisation.

3. Diets containing 138 g protein/kg supplemented with methionine and lysine supported production and food utilisation as effectively as a diet containing 167 g protein/kg.     

The influence of the injection site on the bioavailability of ampicillin and amoxycillin in beagles

Influence of the injection site on bioavailability in dogs was investigated for injections with ampicillin anhydrate or amoxycillin trihydrate suspensions. Firstly, pharmacokinetic parameters were calculated after IV administration of the sodium salts. Then the dogs were injected in the neck (SC), in the lateral thorax region (SC), in the back (IM) and in the thigh (IM), respectively. The most obvious depot effect was seen after subcutaneous injection of ampicillin in the thorax region, though bioavailability seemed to be low. No differences were seen between the injection sites with amoxycillin. For ampicillin SC injection in the neck seems most favourable; for amoxycillin SC injection may be preferred because it is less burdening. Serum concentrations with amoxycillin were higher and persisted longer than with ampicillin. Further investigation is necessary to determine whether this also counts for tissues or focus of infection.     

The influence of nitrogen and defoliation on production and water‐use efficiency of Lolium multiflorum cv. Midmar

Abstract

The objective of this study was to investigate the influence of different nitrogen fertiliser application levels in combination with different defoliation intervals on the dry matter (DM) production and water‐use efficiency of ryegrass (Lolium multiflorum,) cv. Midmar. In a field trial, four nitrogen levels (0, 150, 300 and 450 kg N ha^?1) were combined with five defoliation intervals (every 2, 4, 6 and 8 weeks and twice a season). Soil moisture levels were measured with a neutron hydroprobe and were used to schedule irrigation. The dry matter production, nitrogen‐use efficiency (NUE), plant DM content and water‐use efficiency were influenced (P<0.01) by nitrogen level, defoliation interval, and the interaction between these factors. Total DM production varied from 64 to 20 922 kg DM ha^?1. Dry matter production per unit of applied N decreased with an increase in N level within a defoliation interval. The plant DM content varied from 14.0% to 35.6%. The plant DM content decreased (P<0.05) with increasing N applications, while a lengthening of the defoliation interval resulted in an increase (P<0.05) in the DM content. No difference (P>0.05) was found in total water used when plants received N. Water‐use efficiency varied from 1.2 to 25.8 kg DM ha^?1 mm^?1 water received.     

Matrix transformation alters species-area relationships in fragmented coastal forests

Context

Ecological theory suggests that large habitat fragments should harbour more species than small fragments. However, this may depend on the surrounding matrix. Matrices in fragmented landscapes may either amplify or reduce area effects, which could influence predicted extinctions based on species-area relationships (SARs).

Objective

To determine the influence of matrix type on SARs.

Methods

We surveyed birds within 59 coastal forest fragments in two matrix types, anthropogenic (South Africa) and natural (Mozambique). We classified species as forest specialists or habitat generalists and fitted species-area models to compare how SAR slopes differed among matrix types. We also calculated nestedness and evenness to determine if these varied among matrix type and used logistic regressions to identify species-specific responses to matrix type.

Results

For habitat generalists, SARs were weak within both matrices, while for forest specialists it was strong in the anthropogenic but weak in the natural matrix. In the former, the SAR was similar to those recorded for real islands within archipelagos. Forest specialist assemblages were nested by area within anthropogenic, but not natural matrices. Matrix type did not influence evenness. Area only affected the occurrence of one species when the matrix was natural, compared to 11 species when it was anthropogenic.

Conclusions

Forest specialist bird species conformed to island biogeographic predictions of species loss in forest fragments embedded in anthropogenic, but not natural matrices. Extinctions from small forest fragments might be prevented by conserving natural- or restoring anthropogenic matrices, as well as by increasing forest area.

     

Variance decomposition in the estimation of genetic variance with selected data.

Estimation of genetic variance in populations under selection involves assumptions on base animals. Base animals are often considered unselected and it also has been proposed to treat selected base animals as fixed. The consequences of assumptions on base animals in the estimation of genetic variance in selected populations are not fully understood. Variance decompositions are introduced for simple designs to quantify the differences between models that treat base animals in different ways. Independent contrasts were constructed and REML estimates of variance components were compared for different designs and selection rules. The method shows how selection is accounted for in a complete model and why estimation of variance components can become biased when base animals are treated as fixed.     

Ultrasonographic tissue characterization of equine superficial digital flexor tendons by means of gray level statistics

OBJECTIVE: To correlate quantitative analysis of ultrasonographic images of normal (injury-free) equine superficial digital flexor (SDF) tendons and equine SFD tendons that have pathologic changes with corresponding histologic sections. SAMPLE POPULATION: 4 SDF tendons, 2 of which had various stages of tissue integrity. The 2 ipsilateral tendons were used as points of reference. PROCEDURE: Tendons were mounted in a custom-made device that permitted sequential scanning, transversely and perpendicular to the tendon long axis. At precise steps of 0.5 mm, transverse ultrasonographic images were collected. Subsequently, tendons were fixed and prepared for histologic examination. The following 8 tissue types were discerned: normal young, normal old, necrotic, early granulation, late granulation, early fibrotic, late fibrotic, and scar tissues. In areas of interest, the corresponding ultrasonographic images were selected for gray level statistical analysis. RESULTS: Compared with other tissue types, early-stage granulation tissue was characterized by substantially lower mean gray level and a clearly different histogram. Necrotic tissue had a higher mean gray level, with a virtually normal histogram. In late granulation and early fibrotic tissues, the mean gray level and the histogram could not be discerned from those of normal tendon tissue. The same applied to late fibrotic and scar tissues; mean gray levels were fractionally lower than those of normal tendon tissue with a completely normal histogram. CONCLUSIONS: Although quantification of the transverse ultrasonographic image by use of first-order gray level statistics may be helpful, the method is not sufficiently sensitive to accurately and unequivocally determine the type of tendon tissue. Quantitative analysis should incorporate transverse and longitudinal information.