Minimally invasive patent ductus arteriosus occlusion in 5 dogs

OBJECTIVE: To report a technique for minimally invasive occlusion of patent ductus arteriosus (PDA) and outcome in 5 dogs. STUDY DESIGN: Clinical cases. Animals: Five, 4-6-month-old, dogs with PDA. MATERIALS AND METHODS: Titanium ligating clips were used for PDA closure in all dogs. Three dogs had video-enhanced mini-thoracotomy PDA occlusion. Two other dogs had thoracoscopic PDA occlusion using a custom-designed thoracoscopy clip applicator. RESULTS: Thoracoscopic PDA occlusion was successful in both dogs in which it was attempted. Complete PDA closure was achieved in 4 dogs. Three months after surgery, the largest dog had residual ductal flow that hemodynamically was insignificant. CONCLUSIONS: Although technically demanding, minimally invasive PDA occlusion is a safe and reliable technique in dogs. Preoperative measurement of the diameter of the PDA is crucial to determine if complete closure with metal clips can be achieved. CLINICAL RELEVANCE: Minimally invasive PDA occlusion should be considered as an alternative to occlusion via conventional thoracotomy.     

Assessment of PCR for routine identification of species of the Mycoplasma mycoides cluster in ruminants

DNA amplification techniques offer considerable promise for the identification of Mycoplasma mycoides cluster members. They avoid antigenic cross-reactivity and variability that hamper serological methods. Many sets of primers, specific of these different members and of Mycoplasma putrefaciens, have been proposed. To assess the reliability of some of these PCR tests in routine laboratory diagnostic use, 230 field strains supposed to belong to this group were simultaneously identified by PCR and an antigenic method. The results were well correlated to antigenic identification for M. putrefaciens, but PCR failed to identify respectively 74% and 52% of M. mycoides subsp. mycoides Large Colony type and M. capricolum subsp. capricolum strains. Any identification of M. mycoides subsp. mycoides Small Colony type must be confirmed by two different tests. Difficulties in defining the M. species bovine serogroup 7 were also encountered with both the PCR and immunological methods. The occurrence of putative variable antigen(s) on the mycoplasma surface may explain part of the identification difficulties encountered with the immunological methods.     

Structure of tracheal cytotoxin in complex with a heterodimeric pattern-recognition receptor

Tracheal cytotoxin (TCT), a naturally occurring fragment of Gram-negative peptidoglycan, is a potent elicitor of innate immune responses in Drosophila. It induces the heterodimerization of its recognition receptors, the peptidoglycan recognition proteins (PGRPs) LCa and LCx, which activates the immune deficiency pathway. The crystal structure at 2.1 angstrom resolution of TCT in complex with the ectodomains of PGRP-LCa and PGRP-LCx shows that TCT is bound to and presented by the LCx ectodomain for recognition by the LCa ectodomain; the latter lacks a canonical peptidoglycan-docking groove conserved in other PGRPs. The interface, revealed in atomic detail, between TCT and the receptor complex highlights the importance of the anhydro-containing disaccharide in bridging the two ectodomains together and the critical role of diaminopimelic acid as the specificity determinant for PGRP interaction.     

Sucrose, glucose, and fructose extraction in aqueous carrot root extracts prepared at different temperatures by means of direct NMR measurements

Solutions obtained by heating carrot roots in water (stocks) are widely used in the food industry, but little information is available regarding the metabolites (intermediates and products of metabolism) found in the stock. The effect of treatment temperature and duration on the sugar composition of stocks was investigated directly by quantitative (1)H NMR spectroscopy, to understand the extraction mechanism when processing at 100 degrees C. Stocks prepared at three different temperatures (50, 75, and 100 degrees C) were investigated for up to 36 h. Three sugars (sucrose, glucose, and fructose) were detected and quantified. The concentrations of these three sugars reached a maximum after 9 h when the temperature of treatment was 50 or 75 degrees C. At 100 degrees C, the sucrose concentration reached a maximum after 3 h, whereas the concentration of glucose and fructose was still increasing at that time. Comparison of the kinetic composition of these carrot stocks with that of model sugar solutions leads to the proposal that the changes in stock composition result from sugar diffusion, sucrose hydrolysis, and hydroxymethylfurfural (HMF) formation.     

CD86 molecule is a specific marker for canine monocyte-derived dendritic cells

In this study, canine monocyte-derived dendritic cells (cMo-DC) were produced in presence of canine GM-CSF (cGM-CSF) and canine IL-4 (cIL-4), and they were characterized by their dendritic morphology, MLR functionality and phenotype. We noticed that cMo-DC were labelled with three anti-human CD86 (FUN-1, BU63 and IT2.2 clones), whereas resting and activated lymphocytes or monocytes were not stained. CD86 expression was induced by cIL-4 and was up-regulated during the differentiation of the cMo-DC, with a maximum at day 7. Furthermore, cMo-DC were very potent even in low numbers as stimulator cells in allogeneic MLR, and BU63 mAb was able to completely block the cMo-DC-induced proliferation in MLR. We also observed that cMo-DC highly expressed MHC Class II and CD32, but we failed to determine their maturation state since the lack of commercially available canine markers. Moreover, cMo-DC contained cytoplasmic periodic microstructures, potentially new ultrastructural markers of canine DC recently described. In conclusion, this work demonstrates that the CD86 costimulatory marker is now usable for a better characterization of in vitro canine DC.     

Mapping African Animal Trypanosomosis risk from the sky

In Burkina Faso, African Animal Trypanosomosis (AAT) is still a major hindrance to cattle breeding, especially in the Mouhoun river basin, which was identified as a priority area for tsetse control. The attempt of the present work was to assess the abundance of tsetse flies and AAT risk using remote sensing coupled to field environmental data, along a Mouhoun river section of 234 km long, harbouring an open riverine forest where G. tachinoides Westwood is the predominant tsetse species. The water course was classified into three epidemiological landscapes, corresponding to a "disturbed", "natural" and finally "border" vegetal formation at the interface of the two formers. Using the mean number of infected flies by trap and by day as a risk indicator, the border landscape was found to be 5.4 (1.3-12.0) and 15.8 (4.7-41.6) times more risky than the natural and disturbed ones respectively. These results led to propose that a campaign against tsetse, undertaken by a development project called PAEOB (Projet d'Appui à l'Elevage dans l'Ouest du Burkina Faso), should be focussed on only 34% of the hydrographic network.     

Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system

Background

We previously developed a virus-induced gene silencing (VIGS) vector for cotton from the bipartite geminivirusCotton leaf crumple virus (CLCrV). The original CLCrV VIGS vector was designed for biolistic delivery by a gene gun. This prerequisite limited the use of the system to labs with access to biolistic equipment. Here we describe the adaptation of this system for delivery by Agrobacterium (Agrobacterium tumefaciens). We also describe the construction of two low-cost particle inflow guns.

Results

The biolistic CLCrV vector was transferred into two Agrobacterium binary plasmids. Agroinoculation of the binary plasmids into cotton resulted in silencing and GFP expression comparable to the biolistic vector. Two homemade low-cost gene guns were used to successfully inoculate cotton (G. hirsutum) and N. benthamiana with either the CLCrV VIGS vector or the Tomato golden mosaic virus (TGMV) VIGS vector respectively.

Conclusions

These innovations extend the versatility of CLCrV-based VIGS for analyzing gene function in cotton. The two low-cost gene guns make VIGS experiments affordable for both research and teaching labs by providing a working alternative to expensive commercial gene guns.     

IMAGING DIAGNOSIS–PULMONARY‐TRACHEOBRONCHIAL PROLAPSE IN A NEW CALEDONIAN GIANT GECKO (RHACODACTYLUS LEACHIANUS)

A 3‐year‐old male New Caledonian giant gecko, or Leach's gecko (Rhacodactylus leachianus) presented with acute lethargy and coelomic distention. Findings from survey radiographs and an upper gastrointestinal tract contrast study were consistent with severe aerophagia, a collapsed left lung, and hyperinflation of the right lung due to suspected bronchial obstruction. The gecko was treated with conservative medical management, but was found dead 5 days after presentation. Necropsy findings showed intussusception of the proximal left lung into the left mainstem bronchus and trachea.     

The use of the rapid osmotic fragility test as an additional test to diagnose canine immune-mediated haemolytic anaemia

Background

Diagnosing canine immune-mediated haemolytic anaemia (IMHA) is often challenging because all currently available tests have their limitations. Dogs with IMHA often have an increased erythrocyte osmotic fragility (OF), a characteristic that is sometimes used in the diagnosis of IMHA. Since the classic osmotic fragility test (COFT) is time-consuming and requires specialized equipment, an easy and less labour-intensive rapid osmotic fragility test (ROFT) has been used in some countries, but its diagnostic value has not yet been investigated.This study aimed to evaluate erythrocyte osmotic fragility in dogs with and without IMHA, to compare results of the classic (COFT) and rapid (ROFT) test and to assess the value of the ROFT as diagnostic test for canine IMHA.Nineteen dogs with IMHA (group 1a), 21 anaemic dogs without IMHA (group 1b), 8 dogs with microcytosis (group 2), 13 hyperlipemic dogs (group 3), 10 dogs with lymphoma (group 4), 8 dogs with an infection (group 5) and 13 healthy dogs (group 6) were included.In all dogs, blood smear examination, in-saline auto-agglutination test, Coombs’ test, COFT and ROFT were performed. In the COFT, OF5, OF50 and OF90 were defined as the NaCl concentrations at which respectively 5, 50 and 90% of erythrocytes were haemolysed.

Results

Compared with healthy dogs, OF5 and OF50 were significantly higher in group 1a (P < 0.001) and OF5 was significantly higher in group 3 (P = 0.0266). The ROFT was positive in 17 dogs with IMHA, 10 hyperlipemic dogs, one anaemic dog without IMHA and one healthy dog.

Conclusions

Osmotic fragility was increased in the majority of dogs with IMHA and in dogs with hyperlipidemia, but not in dogs with microcytosis, lymphoma or an infection. Although more detailed information was obtained about the osmotic fragility by using the COFT, the COFT and ROFT gave similar results. The ROFT does not require specialized equipment, is rapid and easy to perform and can be used easily in daily practice. Although, the ROFT cannot replace other diagnostic tests, it may be a valuable additional tool to diagnose canine IMHA.