Genetic variation of foot-and-mouth disease virus isolates recovered from persistently infected water buffalo (Bubalus bubalis)

Genetic variation of foot-and-mouth disease virus (FMDV) isolates, serotype O, recovered serially over a 1-year period from persistently infected buffalos was assessed. The persistent state was established experimentally with plaque-purified FMDV, strain O(1)Campos, in five buffalos (Bubalus bubalis). Viral isolates collected from esophageal-pharyngeal (EP) fluids for up to 71 weeks after infection were analyzed at different times by nucleotide sequencing and T(1) RNase oligonucleotide fingerprinting to assess variability in the VP1-coding region and in the complete genome, respectively. Genetic variation increased, although irregularly, with time after infection. The highest values observed for the VP1-coding region and for the whole genome were 2.5% and 1.8%, respectively. High rates of fixation of mutations were observed using both methodologies, reaching values of 0.65 substitutions per nucleotide per year (s/nt/y) and 0.44s/nt/y for nucleotide sequencing and oligonucleotide fingerprinting, respectively, when selected samples recovered at close time periods were analyzed. The data herein indicate that complex mixtures of genotypes may arise during FMDV type O persistent infection in water buffalos, which can act as viral reservoirs and also represent a potential source of viral variants. These results fit within the quasi-species dynamics described for FMDV, in which viral populations are constituted by related, non-identical genomes that evolve independently from each other, and may predominate at a given time.     

Effects of genetic and environmental factors on chronic lower airway disease in horses

Background : Environment and genetics influence the manifestation of recurrent airway obstruction (RAO), but the associations of specific factors with mild, moderate, and severe clinical signs are unknown.
Hypothesis : We hypothesized that sire, feed, bedding, time outdoors, sex, and age are associated with clinical manifestations of mild, moderate, and severe lower airway disease.
Animals : Direct offspring of 2 RAO-affected Warmblood stallions (F1S1, n = 172; F1S2, n = 135); maternal half-siblings of F1S1 (mHSS1, n = 66); and an age-matched, randomly chosen control group (CG, n = 33).
Methods : A standardized questionnaire was used to assess potential risk factors and to establish a horse owner assessed respiratory signs index (HOARSI 1–4, from healthy to severe) according to clinical signs of lower airway disease.
Results : More F1S1 and F1S2 horses showed moderate to severe clinical signs (HOARSI 3 and HOARSI 4 combined, 29.6 and 27.3%, respectively) compared with CG and mHSS1 horses (9.1 and 6.2%, respectively; contingency table overall test, P < .001). Sire, hay feeding, and age (in decreasing order of strength) were associated with more severe clinical signs (higher HOARSI), more frequent coughing, and nasal discharge.
Conclusions and Clinical Relevance : There is a genetic predisposition and lesser but also marked effects of hay feeding and age on the manifestation of moderate to severe clinical signs, most markedly on coughing frequency. In contrast, mild clinical signs were not associated with sire or hay feeding in our populations.     

Cultivation of three color strains of Gracilaria domingensis in an integrated organic system

Species of the genus Gracilaria are widely distributed around the world, and many are wild harvested and cultivated for phycocolloid production (agar) and human food. This study investigated the growth performance of three color strains of Gracilaria domingensis in an integrated organic system. The three chromatic variants (red, green, and brown) were cultured in a shrimp farm at two depths (surface and 20 cm). The highest relative growth rates were obtained in the first week of cultivation for all three strains studied. Although the growth rates and biomass were slightly higher at a depth of 20 cm, significant differences were not observed at different depths (p < 0.05). The growth of the three strains varied significantly over time (p < 0.001). The maximal growth values obtained were 11.25 ± 1.50% day−1 (red), 12.76 ± 3.96% day−1 (green), and 7.86 ± 1.75% day−1 (brown). The mean growth rates for the whole period were 4.63 ± 6.97, 3.38 ± 9.09, and 2.16 ± 6.61% day−1 for the red, green, and brown strains, respectively. The pigment content (chlorophyll a, phycocyanin, and phycoerythrin) of the three strains studied was higher at a depth of 20 cm, while the maximum electron transport rate (ETRmax) and photosynthetic efficiency were higher at the surface. The high growth rates obtained in this study demonstrate the great potential of this species for integrated organic farming. Moreover, the ecological success shown by the red strain suggests that this strain is the most suitable for integrated cultivation.     

Total Mercury Bioaccumulation in Tissues of Carnivorous Fish (Micropogonias furnieri and Cynoscion acoupa) and Oysters (Crassostrea brasiliana) from Sepetiba Bay,Brazil

Total mercury (Hg) levels were determined by cold vapor atomic absorption spectrophotometry (CVAAS) in tissues of two species of carnivorous fishes and in the soft tissue of oysters caught in Sepetiba Bay, Brazil. Hg was distributed unevenly in the organs and tissues of the fish; the mean Hg content was highest in the kidneys (0.048 μg/g wet weight) and lowest in the gills (0.004 μg/g w.w.). Mean Hg levels were higher in the oysters (0.052 μg/g w.w.) than in the fish (0.022 μg/g w.w). The total Hg concentration did not exceed the maximum concentration for food allowed by Brazilian legislation in any of the fish and oyster samples analyzed. Moreover, the Hg concentrations observed were all within the maximum level for fish recommended by the Food and Agriculture Organization (FAO; 0.5 μg/g w.w.).     

Desmoglein-1 is a minor autoantigen in dogs with pemphigus foliaceus

The majority of human patients with pemphigus foliaceus (PF) have circulating IgG autoantibodies that target conformational epitopes on the desmosomal cadherin desmoglein-1 (dsg1). Limited studies using immunoblot techniques suggested that the principal autoantigen in dogs with PF might also be dsg1. It was the objective of this study to test this hypothesis. A comprehensive survey of canine PF sera was conducted using a novel screening strategy that detects conformational epitopes. This method consists of the ectopic expression of canine dsg1 at the surface of human 293T epithelial kidney cells and their live screening, i.e. prior to fixation. Out of seven control human PF sera that bound to canine epidermis, three (57%) contained IgG autoantibodies that recognized ectopically expressed canine dsg1 with a membrane and punctate pattern. Out of 83 canine PF sera only five (6%) contained IgG that recognized canine dsg1. Consistent with findings for human PF sera obtained in this study, autoantibody binding was conformation- and glycosylation-dependent as demonstrated by calcium chelation with EDTA and tunicamycin or wheat germ agglutinin treatment, respectively. In conclusion, these studies establish canine dsg1 as a minor autoantigen for canine PF. Antigenic epitopes appear to be conformation- and glycosylation-dependent.     

Characterization of the inflammatory infiltrate and cytokine expression in the skin of horses with recurrent urticaria

Background – Recurrent urticaria (RU) is a common skin disease of horses, but little is known about its pathogenesis. Hypothesis/Objective – The aim of this study was to characterize the inflammatory cell infiltrate and cytokine expression pattern in the skin of horses with RU. Animals – Biopsies of lesional and nonlesional skin of horses with RU (n = 8) and of skin from healthy control horses (n = 8) were evaluated. Methods – The inflammatory cell infiltrate was analysed by routine histology. Immunohistochemistry was used to identify T cells (CD3), B cells (CD79), macrophages (MAC387) and mast cells (tryptase). Expression of T‐helper 2 cytokines (interleukins IL‐4, IL‐5 and IL‐13), a T‐helper 1 cytokine (interferon‐γ), IL‐4 receptor α and thymic stromal lymphopoietin was assessed by quantitative RT‐PCR. Results – In subepidermal lesional skin of RU‐affected horses, increased numbers of eosinophils (P ≤ 0.01), CD79‐positive (P ≤ 0.01), MAC387‐positive (P ≤ 0.01) and tryptase‐positive cells (P ≤ 0.05) were found compared with healthy horses. Subepidermal lesional skin of RU‐affected horses contained more eosinophils (P ≤ 0.05) and tryptase‐positive cells (P ≤ 0.05) compared with nonlesional skin. There was no significant difference in infiltrating cells between nonlesional skin and skin of healthy horses. Expression of IL‐4 (P ≤ 0.01), IL‐13 (P ≤ 0.05), thymic stromal lymphopoietin (P ≤ 0.05) and IL‐4 receptor α (P ≤ 0.05) was increased in lesional skin of RU‐affected horses compared with control horses. Expression of IL‐4 was higher (P ≤ 0.05) in lesional compared with nonlesional RU skin. Conclusions and clinical importance – Analysis of cytokine expression and inflammatory infiltrate suggests that T‐helper 2 cytokines, eosinophils, mast cells and presumptive macrophages play a role in the pathogenesis of equine RU.