Detection of Leishmania spp. and associated inflammation in ocular‐associated smooth and striated muscles in dogs with patent leishmaniosis

Objective Canine leishmaniosis is a disease characterized by the wide distribution of the parasite throughout the tissues of the host. The purpose of this study was to describe the presence of Leishmania spp. and associated inflammation in ocular‐associated muscles of dogs with patent leishmaniosis. Procedures Smooth muscles (iris dilator muscle, iris sphincter muscle, ciliary muscle, Müller muscle, smooth muscle of the periorbita and smooth muscle of the nictitating membrane) and striated muscles (orbicularis oculi muscle, obliquus dorsalis muscle and dorsal rectus muscle) were evaluated. Routine staining with hematoxylin and eosin and immunohistochemistry to detect Leishmania spp. were performed on tissue sections. Results Granulomatous inflammation was seen surrounding muscular fibers and was composed mainly of macrophages with scattered lymphocytes and plasma cells. This infiltrate could be seen in 52/473 (10.99%) samples of smooth muscle and 36/142 (25.35%) samples of striated muscle. Parasites were detected in 43/473 (9.09%) samples of smooth muscle and in 28/142 (19.71%) samples of striated muscle. Conclusions To the authors’ knowledge, this is the first report assessing the presence of Leishmania spp. and associated infiltrate in intraocular, extraocular and adnexal smooth and striated muscles. The inflammation present in those muscles could contribute to clinical signs already described, such as blepharitis, uveitis, and orbital cellulitis.     

Altering the substrate specificity of Candida rugosa LIP4 by engineering the substrate-binding sites

Candida rugosa (formerly Candida cylindracea) lipase (CRL) is an important industrial enzyme that is widely used in biotechnological applications such as the production of fatty acids and the synthesis of various esters. CRL comprises at least seven isozymes (LIP1-LIP7), which share a similar amino acid sequence but with different specificities for substrates. Previously, LIP4 was reported to have higher esterase activity toward long acyl-chain ester and lower lipase activity toward triglycerides. A296 and V344 of LIP4 were predicted to play decisive roles in its substrate specificity. In this study, site-specific saturation mutagenesis has been employed to study the substrate specificity of LIP4. Point mutations were separately introduced into A296 and V344 positions using degenerate primer sets containing 32 codons to generate two libraries of variants. LIP4 variants were heterologously expressed in the yeast Pichia pastoris. A specific plate assay was used to identify lipase-producing P. pastoris clones in a medium containing tributyrin. LIP4 variants with high activity toward short fatty acyl-chain triglyceride (tributyrin) were screened. Specificity analysis and biochemical characterization indicated that the recombinant variants A296I, V344Q, and V344H had properties remarkably different from those of wild-type LIP4. All three variant enzymes had significantly higher specific activities toward tributyrin than LIP4. In addition to short-chain triglyceride, A296I and V344Q also improved hydrolytic activities of triglycerides toward medium- and long-chain triglycerides tested. The results suggested that A296 played an important role in lipase activity and high-temperature dependence of LIP4, whereas it had no effect on the chain-length specificity in lipolytic reaction. The V344 residue had a significant effect on the substrate chain-length specificity of LIP4.     

Body development, carcass, and meat quality of confined lambs fed increasing levels of whole rice meal

The objective of this trial was to evaluate the effect of whole rice meal (WRM) inclusion in the concentrate upon body development, carcass traits, and meat quality of lambs. Twenty-four castrated lambs with an average initial body weight of 17.90?±?2.72 kg were randomly blocked according to two genetic groups (Corriedale and Texel by Corriedale crossbreds). Three isocaloric (11.3 MJ/kg of metabolizable energy) and isonitrogenous (17 % crude protein) diets were offered to the animals for 74 days. Diets consisted of 40 % forage and 60 % concentrate diet, on a dry matter basis, with 0, 15, or 30 % of WRM inclusion into the concentrate. Body growth (after slaughter), carcass, and meat traits were evaluated on each animal. Results obtained indicated that genotype did not affect body growth, carcass, and meat traits except for yellowness. No significant interaction between diet and genotype were detected. Inclusion of up to 30 % WRM did not significantly (P?>?0.05) affect body growth, carcass, and meat traits, except for meat color. Meat luminosity progressively increased (36.32?+?0.055X) while redness (15.13???0.03X) decreased with the inclusion of WRM in the diet, but still remained within acceptable values. The study indicates that WRM may be included up to 30 % in the concentrate replacing corn without adverse effects upon body development, carcass traits, and meat quality of lambs.     

Subependymal Reaction Secondary to Choroid Plexus Papilloma in a Horse

Choroid plexus papilloma (CPP) is a rare, benign, intracranial neoplasm. The present report described a choroid plexus papilloma of the fourth ventricle with unusual proliferative reaction of ependymal/subependymal cells in a stallion. A single, soft neoformation, red-gray in color, with an irregular surface, resembling a cauliflower, expanding from the fourth ventricle to the cerebellar peduncles was observed. Microscopically, the neoformation was composed of a single layer of simple cuboidal to columnar epithelium with a well-vascularized papillary-arboriform pattern. Multifocal subependymal aggregates forming rosettes were observed near the mesencephalic aqueduct (MA). Glial-fibrillary-acidic-protein (GFAP) was markedly positive only in the neuropil adjacent to the proliferative areas of the MA. Ependymal/subependymal reactions secondary to CPP have never been described in equine or human neuropathology. The capabilities of ependymal cells to regenerate and eventually proliferate are controversial and unclear and, thus, require additional investigation to understand the adaptive mechanisms of this rare pathogenic condition.     

Growth, stress response and free amino acid levels in Senegalese sole (Solea senegalensis Kaup 1858) chronically exposed to exogenous ammonia

Stressful husbandry conditions are likely to affect growth and amino acid metabolism in fish. In this study, chronic ammonia exposure was used to test the effects of a stressor on growth and amino acid metabolism of Senegalese sole juveniles. The fish were exposed for 52 days to 11.6 mg L⁻¹ [low‐TAN (L‐TAN)] or 23.2 mg L⁻¹ [high‐TAN (H‐TAN)] of total ammonia nitrogen (TAN), or to 0 mg L⁻¹ (Control). Growth in L‐TAN groups was slightly but significantly different from the Control groups [relative growth rate (RGR=0.35±0.13 and 0.52±0.23% day⁻¹ respectively)]. In H‐TAN groups, growth was severely affected (RGR=0.01±0.13% day⁻¹). Stress parameters (plasma cortisol and glucose) showed slight or no significant differences between treatments. Plasma free amino acid (FAA) concentrations were affected after H‐TAN treatment. Increases in glutamine and aspartate concentrations in H‐TAN fish suggest alterations in amino acid metabolism related to nitrogen excretion processes. Some of the changes in FAA concentrations also suggest mobilization to energy supply and synthesis of metabolites related to stress response. Therefore, Senegalese sole seem to adapt to the L‐TAN concentration tested, but the H‐TAN concentration reduced growth and affected amino acid metabolism.     

Benthic microbial indicators of fish farm impact in a coastal area of the Tyrrhenian Sea

We studied the impact of organic loads due to the biodeposition of a fish farm in a non-impacted coastal area of the Tyrrhenian Sea (Western Mediterranean). Sediment chemistry and benthic microbial community were investigated from July 1997 to February 1998 on monthly basis at two stations: one was located under the fish farm, while the second was about 1 km away, and served as a reference site. The presence of the cage induced rapid changes in the benthic conditions: the sediments were rapidly (after 6 weeks) reduced. A significant accumulation of biopolymeric carbon was observed beneath the cage both 2 weeks after the initial cage deployment, on non-impacted sediments, and 5–7 months after, and appeared to be related to the fish farm production cycle. The density of microbial communities beneath the fish farm increased only during the first month of farming activity (July) and partially 7 months after cage disposal (i.e. in January), when there was a significant sediment organic enrichment. Additional evidence of rapid impact of the fish farm on the benthic bacterial communities is provided by the increase in the numbers of autofluorescent microbial cells. The photosynthetic eukaryotic cells displayed a highly reduced contribution to total autofluorescent microbial density, indicating that their decrease is related with biopolymeric carbon accumulation, and probably was due also to “shadow effect” induced by suspended material, coming from the fish farm, on the environment below. We propose here, to apply the ratio of culturable heterotrophic bacteria to microbial direct counts (CFU/MDC) to detect fish farm impact. In cage sediments, organic enrichment and the consequent modification of the characteristics of the benthic environment, determined an increase in aerobic heterotrophic bacteria and vibrio density indicating that they are efficient colonizers of organic-rich sediments. Densities of Escherichia coli and Enterococci were not significantly higher than in the reference site, and are likely of terrestrial origin. Thus they cannot be used as specific indicators of fish farm impact.