Cytological, Immunocytochemical and Ultrastructural Study of the Adenohypophyseal Pars Distalis of the Kid (Capra hircus): The TSH cell

The structure and ultrastructure of the adenohypophyseal pars distalis in kids of the Murciano-granadiana breed (18 males and 12 females), aged 2-3 months and with an average carcass weight of 8 kg has been studied. Techniques of staining (Tetrachrome Herlant's, and Cleveland-Wolfe) and histochemistry (PAS, PAS-OG and BA-PAS-OG) contrasted with immunolabelling (PAP) have been used. In addition an ultrastructural study has been made and nucleus and cytoplasm, secretory granules, mitochondria and lysosomes have been measured with a semiautomatic image analyzer (IBAS-1). TSH cells are found in sagittal section in the anterior area and in an antero-caudodorsal band, and transversally in the ventral and medial region; they are 6% and their average size is 14.15 microns. Ultrastructurally they are a single cellular type with spherical granules whose size is 195 nm in males and 149 in females; these granules are characterized by a clear halo and a variable electronic density. The rough endoplasmic reticulum is found as slightly enlarged vesicles with a homogeneous and moderately electro-dense content or in concentric stratifications.     

Accuracy of asymmetry indices of ground reaction forces for diagnosis of hind limb lameness in dogs

OBJECTIVE: To determine the accuracy of asymmetry indices of ground reaction forces (GRF) for diagnosis of hind limb lameness in dogs. ANIMALS: 36 healthy dogs and 13 dogs with naturally acquired cranial cruciate ligament rupture or hip dysplasia. PROCEDURES: Lameness for affected dogs ranged from not detectable to minor and constant. While dogs trotted on an instrumented treadmill, GRF variables were recorded and analyzed with asymmetry indices. Each index was tested for its ability to discriminate between healthy and affected dogs. Combinations of several indices were also assessed. RESULTS: Vertical force variables had better accuracy than craniocaudal force variables. Peak vertical force was the most accurate variable. Partial asymmetry during trotting was detected in healthy dogs. A multivariate approach that used peak vertical force and maximal rising slope yielded the optimum combination to distinguish between healthy and affected dogs. In addition, sensitivity of 92% or specificity of 95% may be achieved with 2 cutoff values while simultaneously maintaining specificity or sensitivity, respectively, at > 85%. CONCLUSIONS AND CLINICAL RELEVANCE: Asymmetry indices of GRFs were accurate for detection of hind limb lameness in dogs. This is particularly relevant for study designs in which only a single gait evaluation is possible.     

Primary orthostatic tremor in Great Danes

Two Great Danes were presented for an investigation of 6-month histories of incessant tremor of all 4 limbs and trunk occurring only when the dogs were standing at rest. Dog 1 was a 2-year-old intact female harlequin Great Dane living in the UK, and dog 2 was a 2-year-old castrated male fawn Great Dane living in the United States. The owners described both dogs as having constant quivering of the limbs while standing, suggestive of an involuntary dance, since the age of 18 months. The intensity of the tremors was reported in both dogs to have steadily increased since onset. In dog 1, tremors were noticed to be more pronounced in the fore limbs when she was eating and more pronounced in the hind limbs when she was trying to sit. The owner of dog 1 also reported that the dog could no longer assume a position of recumbency without manual assistance because of the severity of the tremors. Intermittently, tremors were reported to involve the facial musculature of dog 2. Vaccinations were current, and the owners of both dogs did not report any previous medical history, exposure to natural or manufactured toxins, or concurrent treatment. No familial relationship was identified between these 2 dogs.     

Porcine CD18 mediates Actinobacillus pleuropneumoniae ApxIII species-specific toxicity

Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, produces Apx toxins that are recognized as major virulence factors. Recently, we showed that ApxIIIA-cytotoxic activity specifically targets Sus scrofa leukocytes. Since both LtxA from Aggregatibacter actinomycetemcomitans (aggressive periodontitis in humans) and LktA from Mannheimia haemolytica (pneumonia in ruminants) share this characteristic, respectively towards human and ruminant leukocytes, and because both use the CD18 subunit to interact with their respective LFA-1, we hypothesized that ApxIIIA was likely to bind porcine CD18 to exercise its deleterious effects on pig leukocytes. A β ₂−integrin-deficient ApxIIIA-resistant human erythroleukemic cell line was transfected either with homologous or heterologous CD11a/CD18 heterodimers using a set of plasmids coding for human (ApxIIIA-resistant), bovine (-resistant) and porcine (-susceptible) CD11a and CD18 subunits. Cell preparations that switched from ApxIIIA-resistance to -susceptibility were then sought to identify the LFA-1 subunit involved. The results showed that the ApxIIIA-resistant recipient cell line was rendered susceptible only if the CD18 partner within the LFA-1 heterodimer was that of the pig. It is concluded that porcine CD18 is necessary to mediate A. pleuropneumoniae ApxIIIA toxin-induced leukolysis.     

Application of a putative fatty-acid binding protein to discriminate serologically the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from other Meloidogyne species

Two major proteins, Mcf-A67 and Mcf-B66, were identified by mini two-dimensional polyacrylamide gel electrophoresis in order to distinguish the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from eight other species. These quarantine proteinic markers have been microsequenced after enzymatic digestion. The internal amino acid sequences exhibit similarities to members of a family of low molecular weight intracellular lipid-binding proteins. Moreover, to explore a simple, rapid, and inexpensive way to identify the two quarantine nematodes, dot blot hybridizations were performed using an antiserum (A67) produced from the longest amino-acid sequence of the protein Mcf-A67. Although several proteins stained on the M. chitwoodi and M. fallax western blot membranes, the two nematodes were easily distinguished from other root-knot nematodes, on dot blot assays with soluble proteins extracted from a single female. Because of its specificity and sensitivity, the use of the A67 antiserum to improve the diagnosis of the two European quarantine root-knot nematodes is discussed.     

Abundant PrP(CWD) in tonsil from mule deer with preclinical chronic wasting disease.

A monoclonal antibody dot-blot assay was used to evaluate detergent lysates of tonsil tissue from mule deer to detect PrP(CWD), the marker for the cervid transmissible spongiform encephalopathy chronic wasting disease (CWD). Samples of formalin-fixed brain and tonsil tissues from mule deer were examined for PrP(CWD) using immunohistochemistry (IHC) with Mab F99/97.6.1, the gold standard for diagnosis of preclinical CWD. The contralateral tonsil from each of the 143 deer was prepared for confirmatory IHC and as a 10% (wt/vol) detergent lysate without purification or enrichment steps for monoclonal antibody dot-blot assay. PrP(CWD) was detected by dot-blot assay in 49 of 50 samples considered positive by IHC. Forty-eight of the positive samples were evaluated with a quantitative dot-blot assay calibrated with recombinant PrP. Tonsillar PrP(CWD) concentrations ranged from 34 to 1,188 ng per 0.5 mg starting wet weight of tissue. The abundant PrP(CWD) in mule deer tonsil will facilitate development and validation of high-throughput screening tests for CWD in large populations of free-ranging deer.     

Rapid seedlings regeneration from seeds and vegetative propagation with sucker and rhizome of Eremospatha macrocarpa (Mann & Wendl.) Wendl and Laccosperma secundiflorum (P. Beauv.) Kuntze

To develop efficient seedling production methods for Laccosperma secundiflorum and Eremospatha macrocarpa, a study was conducted to examine regeneration using offsets combined with several physical and chemical treatments of seeds. Offsets categorized into small, medium and large diameters, were planted in three conditions: shaded and open nursery, and greenhouse. We tested sucker from E. macrocarpa, and sucker and rhizome from L. secundiflorum. For both species, high viability percentage (ranging from 55% to 100%) were observed for small and medium suckers planted in shaded nursery and greenhouse, against less than 49% for sucker planted in open nursery. The mean seedling emergence times were estimated to 84, 77 and 75 days after planting (DAP) for small, medium and large sucker of L. secundiflorum, respectively under open nursery condition, and 76, 75, 95 DAP for small, medium and large suckers of the same species, respectively in shaded condition. Greenhouse has a significant positive effect on E. macrocarpa seedlings emergence time. For this species, the mean seedling emergence times were estimated to 43 DAP for small sucker and 76, 93 DAP for medium and large suckers. No seedling was obtained from rhizome planted in all the growing conditions tested. Concerning seed dormancy breaking, germination percentages and rates were determined for 13 treatments. The best treatments were pre-soaking unscarified seeds for 4 days in 1.01 g l⁻¹ and 0.10 g l⁻¹ KNO₃, with 79% and 68% of germination, respectively and in 3.46 × 10⁻³ g l⁻¹ GA₃ for 68% of germination. These methods are suggested to improve germination of L. secundiflorum seeds. Successful and recommended methods for E. macrocarpa are pre-soaking scarified seeds in 3.46 × 10⁻³ g l⁻¹ and 3.46 × 10⁻⁴ g l⁻¹ GA₃, 96% and 94% of germination, respectively. Dormancy, probably a combination of mechanical and chemical dormancy, is present in the two species.     

Diuron mineralisation in a Mediterranean vineyard soil: impact of moisture content and temperature

BACKGROUND: The diuron‐mineralising ability of the microbiota of a Mediterranean vineyard soil exposed each year to this herbicide was measured. The impact of soil moisture and temperature on this microbial activity was assessed. RESULTS: The soil microbiota was shown to mineralise diuron. This mineralising activity was positively correlated with soil moisture content, being negligible at 5% and more than 30% at 20% soil moisture content. According to a double Gaussian model applied to fit the dataset, the optimum temperature/soil moisture conditions were 27.9 °C/19.3% for maximum mineralisation rate and 21.9 °C/18.3% for maximum percentage mineralisation. The impact of temperature and soil moisture content variations on diuron mineralisation was estimated. A simulated drought period had a suppressive effect on subsequent diuron mineralisation. This drought effect was more marked when higher temperatures were used to dry (40 °C versus 28 °C) or incubate (28 °C versus 20 °C) the soil. The diuron kinetic parameters measured after drought conditions were no longer in accordance with those estimated by the Gaussian model. CONCLUSION: Although soil microbiota can adapt to diuron mineralisation, its activity is strongly dependent on climatic conditions. It suggests that diuron is not rapidly degraded under Mediterranean climate, and that arable Mediterranean soils are likely to accumulate diuron residues. Copyright © 2010 Society of Chemical Industry