Endotoxin-induced changes in expression of cyclooxygenase isoforms in the lamellar tissue of extracorporeally haemoperfused equine limbs

Angiogenesis and sepsis-related equine laminitis have several features in common. Both events can be induced by endotoxin (lipopolysaccharide— LPS) and both are associated with increased expression of the enzyme cyclooxygenase (COX), of which two isoforms (COX-1 and COX-2) exist. To examine the causal relationship between LPS exposure and COX expression and to investigate the tissue distribution of COX in the LPS-exposed tissue, the technique of extracorporeal haemoperfusion of isolated equine forelimbs was utilized. Perfusion was performed for 10 hr under physiological conditions (control-perfused limbs, n = 5) and with addition of 80 ng/L of endotoxin (LPS-perfused limbs; n = 5). After perfusion, samples of lamellar tissue were collected from the dorsal aspect of the hoof wall. Additional control samples were collected from three non-perfused limbs. Immunohistochemical analysis was performed using antibodies against COX-1 and COX-2, and intensity of immunohistochemical staining was scored for each isoform. In the lamellar tissue of control- and LPS-perfused limbs, there was no significant difference in COX-1 staining intensity and distribution, whereas COX-2 expression was significantly increased in LPS-perfused limbs (especially in endothelial cells, fibroblasts and intravasal leucocytes as well as in epidermal basal cells at the base of the primary epidermal lamellae). These results suggest that COX-2 and its metabolites are involved in the initiation of pathological changes seen in sepsis-associated events such as sepsis-related laminitis. In such cases, COX-2 could therefore be an important therapeutic target; however, early therapy may be required as increase in COX-2 expression occurs within 10 hr after LPS exposure.     

Development of a murine ocular posterior segment explant culture for the study of intravitreous vector delivery

The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.     

Primary bilateral corneal nerve sheath neoplasm in a dog

A 15‐year‐old, neutered male, Shih Tzu cross developed progressive corneal stromal thickening and vascularization of the right eye, and 5 months later, of the left eye. Both eyes became blind due to extensive corneal opacification and were enucleated. Light microscopic examination revealed a diffuse corneal infiltrate of neoplastic mesenchymal cells, and immunohistochemistry revealed diffuse cytoplasmic vimentin immunoreactivity and variable cytoplasmic and nuclear immunoreactivity for S100 in the neoplastic cells. Transmission electron microscopy revealed desmosomes between contiguous cells, thread‐like cytoplasmic processes coated with basement membrane, extracellular bundles of collagen, and axonal degeneration consistent with features of a nerve sheath neoplasm. This is the first report of primary, bilateral corneal nerve sheath sarcoma in a canine.     

Amylopectin structure and crystallinity explains variation in digestion kinetics of starches across botanic sources in an in vitro pig model

Background: Starch is the main source of energy in commonly used pig diets.Besides effects related to the extent of starch digestion,also several effects related to variation in digestion rate have recently been demonstrated in non-ruminants.Different rates of starch digestion in animals and in in vitro models have been reported,depending on the botanic origin of starch.Starches from different botanic sources differ widely in structural and molecular properties.Predicting the effect of starch properties on in vitro digestion kinetics based on existing literature is hampered by incomplete characterization of the starches,or by a selective choice of starches from a limited number of botanic sources.This research aimed to analyse the relationships between starch properties and in vitro digestion kinetics of pure starches isolated from a broad range of botanic origins,which are used in non-ruminant diets or have a potential to be used in the future.Therefore we studied starch digestion kinetics of potato,pea,corn,rice,barley,and wheat starches,and analysed the granule diameter,number of pores,type and amount of crystalline structure,amylose content and amylopectin side-chain length of all starches.Results: Multivariate analysis revealed strong correlations among starch properties,leading us to conclude that effects of most starch characteristics are strongly interrelated.Across all analysed botanic sources,crystalline type and amylopectin chain length showed the strongest correlation with in vitro digestion kinetics.Increased percentages of A–type crystalline structure and amylopectin side chains of DP 6–24 both increased the rate of digestion.In addition,within,but not across,(clusters of) botanic sources,a decrease in amylose content and increase in number of pores correlated positively with digestion kinetics.Conclusion: The type of crystalline structure and amylopectin chain length distribution of starch correlate significantly with digestion kinetics of starches across botanic sources in an in vitro pig model.Variation in digestion kinetics across botanic sources is not additively explained by other starch properties measured,but appears to be confined within botanical sources.     

Evaluation of the success of medical management for presumptive thoracolumbar intervertebral disk herniation in dogs

Objective— To determine the success of medical management of presumptive thoracolumbar disk herniation in dogs and the variables associated with treatment outcome.
Study Design— Retrospective case series.
Animals— Dogs (n=223) with presumptive thoracolumbar disk herniation.
Methods— Medical records from 2 clinics were used to identify affected dogs, and owners were mailed a questionnaire about success of therapy, recurrence of clinical signs, and quality of life (QOL) as interpreted by the owner. Signalment, duration and degree of neurologic dysfunction, and medication administration were determined from medical records.
Results— Eighty-three percent of dogs (185/223) were ambulatory at initial evaluation. Successful treatment was reported for 54.7% of dogs, with 30.9% having recurrence of clinical signs and 14.4% classified as therapeutic failures. From bivariable logistic regression, glucocorticoid administration was negatively associated with success ( P =.008; odds ratio [OR]=.48) and QOL scores ( P =.004; OR=.48). The duration of cage rest was not significantly associated with success or QOL. Nonambulatory dogs were more likely to have lower QOL scores ( P =.01; OR=2.34).
Conclusions— Medical management can lead to an acceptable outcome in many dogs with presumptive thoracolumbar disk herniation. Cage rest duration does not seem to affect outcome and glucocorticoids may negatively impact success and QOL. The conclusions in this report should be interpreted cautiously because of the retrospective data collection and the use of client self-administered questionnaire follow-up.
Clinical Relevance— These results provide an insight into the success of medical management for presumptive thoracolumbar disk herniation in dogs and may allow for refinement of treatment protocols.     

DNA barcoding reveals blend of silver catfish Rhamdia species from fish farms in Southern Brazil

Rhamdia quelen is the most produced native freshwater fish in fish farms in South Brazil. Until recently, Rhamdia branneri and Rhamdia voulezi were synonyms of R. quelen, and all the species are commercialized as silver catfish (locally called jundiá or bagre sapo) by the aquaculture industry. In fact, because these species are morphologically very similar, interspecific crosses easily might occur in fish farming. We employed standard DNA barcoding to identify jundiá molecular operational taxonomic units (MOTUs) in fish cultivated and commercialized in the industry and in possible escapees in the natural environment in southern Brazil. We analysed 48 individuals from six fish farms and 48 individuals from three rivers (Uruguay, Benedito Novo and Itapocu Rivers) using the mitochondrial cytochrome oxidase subunit I (COI). Four MOTUs were identified based on the estimated optimum threshold (OT = 0.77), and these MOTUs were concordant with Bayesian Inferece (BI) and Neighbour‐Joining (NJ) trees. Our results support the existence of at least three species in our dataset: R. branneri, R. voulezi, and R. quelen 1 and R. quelen 2. The interspecific genetic divergence ranged from 1.1% to 5.1% (mean = 3.5%), and the intraspecific distance ranged from 0% to 1.4% (mean = 0.24%). The presence of cultivated fish in the Uruguay and Benedito Novo Rivers provides evidence of genetic contamination in native populations. These results show the need to regulate aquaculture activities and to characterize the species and commercial lineages of silver catfish that are cultivated in South Brazil.     

Fish assemblages in Atlantic Forest streams: the relative influence of local and catchment environments on taxonomic and functional species

Streams are under environmental pressures acting at different scales that influence the ecological organisation of their fish assemblages. However, the relative influence of the different scale‐related variables on assemblage composition and function is poorly understood. We evaluated the importance of local‐ and catchment‐scale environmental variables, as well as the spatial structure of the sampling sites, in shaping fish assemblages in Atlantic Forest streams. Local‐scale variables were those measured at the sampling sites, describing the local habitat conditions (e.g. depth, substrate type, altitude). Catchment‐scale variables were those integrating the upstream landscape of the sampling sites (e.g. catchment land use). Spatial distances were calculated from watercourse distance using principal coordinates of neighbour matrices. Altogether, 28 local and seven catchment variables were initially subjected to two processes of eliminating co‐linearity. Redundancy analysis was applied to the three matrices (spatial, local and catchment) to quantify the variance in the structure of the fish assemblages explained by each matrix. Local variables explained more variability in both taxonomic and functional assemblage structure, than catchment and spatial variables. Local variables also changed along the longitudinal gradient, which consequently influenced fish assemblage structure. This pattern was also influenced by anthropogenic alteration and non‐native species, which were more abundant in downstream sites. These results highlight the need to assess Atlantic Forest streams under different environmental scales, especially through the use of quantitative local‐scale metrics, and to consider the effects of longitudinal patterns in structuring fish assemblages when developing and implementing monitoring programmes, impact studies and conservation plans.