Purification and renaturation of membrane neuraminidase from Haemophilus parasuis

Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems. Co-factors for and the pathogenesis of H. parasuis disease are not defined. One of the potential virulence factors of H. parasuis is its neuraminidase (sialidase). While purifying the H. parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels. The H. parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82 kilodalton (kDa). The H. parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H. parasuis cell protein while retaining over 90% of the neuraminidase activity. Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein.     

Sequence variants in SLITRK1 are associated with Tourette's syndrome

Tourette's syndrome (TS) is a genetically influenced developmental neuropsychiatric disorder characterized by chronic vocal and motor tics. We studied Slit and Trk-like 1 (SLITRK1) as a candidate gene on chromosome 13q31.1 because of its proximity to a de novo chromosomal inversion in a child with TS. Among 174 unrelated probands, we identified a frameshift mutation and two independent occurrences of the identical variant in the binding site for microRNA hsa-miR-189. These variants were absent from 3600 control chromosomes. SLITRK1 mRNA and hsa-miR-189 showed an overlapping expression pattern in brain regions previously implicated in TS. Wild-type SLITRK1, but not the frameshift mutant, enhanced dendritic growth in primary neuronal cultures. Collectively, these findings support the association of rare SLITRK1 sequence variants with TS.     

Effect of focused and radial extracorporeal shock wave therapy on equine bone microdamage

OBJECTIVES : To determine whether bone microcracks are altered after application of focused and radial extracorporeal shock wave therapy (ESWT) to the equine distal limb. STUDY DESIGN : An ex vivo experimental model. SAMPLE POPULATION : A contralateral limb specimen was obtained from 11 Thoroughbred racehorses with a unilateral catastrophic injury. Distal limb specimens were also obtained from 5 non-racing horses. METHODS : Three separate skin-covered bone segments were obtained from the mid-diaphysis of the metacarpus (MC3) or metatarsus (MT3). Focused (9,000 shockwaves, 0.15 mJ/mm2, 4 Hz) and radial (9,000 shockwaves, 0.175 mJ/mm2, 4 Hz) ESWT treatments were randomized to the proximal and distal segments and the middle segment was used as a treatment control for pre-existing microcracks. After treatment, bone specimens were bulk-stained with basic fuchsin and microcracks were quantified in transverse calcified bone sections. RESULTS : ESWT had small but significant effects on microcracks. Microcrack density (Cr.Dn) and microcrack surface density (Cr.S.Dn) were increased after focused ESWT, whereas Cr.Le was increased after radial ESWT. In racing Thoroughbreds, Cr.Le increased with increased number of races undertaken. Cr.Dn and Cr.S.Dn were not significantly influenced by the number of races undertaken. CONCLUSION : ESWT has small but significant effects on bone microcracking ex vivo. CLINICAL RELEVANCE : These preliminary data suggest that ESWT has the potential to increase bone microcracking in equine distal limb bone in vivo. Such effects may be more pronounced in Thoroughbreds that are actively being raced, because in vivo microcracking increases with increased number of races undertaken.     

Wetland effects on lake water quality in the Minneapolis/St. Paul metropolitan area

A method developed to evaluate the cumulative effect of wetland mosaics on water quality was applied to 33 lake watersheds in the seven-county region surrounding Minneapolis-St. Paul, Minnesota. A geographic information system (GIS) was used to record and measure landscape variables derived from aerial photos. Twenty-seven watershed land-use and land-cover variables were reduced to eight principal components which described 85% of the variance among watersheds. Relationships between lake water quality variables and the first six principal components plus an index of lake mixis were analyzed through stepwise multiple regression analysis. A combination of three landscape components (wetland/watershed area, agriculture/wetlands, and forest/soils components) explained 49% of the variance in a trophic state index, even though most of the lakes examined were already highly eutrophic, and thus were influenced by internal loading. The regression equations explained a range of 14 to 76% of the variation in individual water quality variables. Forested land-use was associated with lower lake trophic state, chloride, and lead. High lake trophic state was associated with agricultural land-use and with wetland distance from the lake of interest. The extent of wetlands was associated with low total lead and high color in lakes downstream. Wet meadows or herbaceous, seasonally-flooded wetlands contributed more to lake water color than did cattail marshes.     

The role of flow dependency and water availability in fish assemblage homogenization in tributaries of the Chattahoochee River,Alabama, USA

The availability of water is becoming more variable as flow regime alterations intensify both locally and on a global scale. This pattern has been observed in the tributaries of the central Chattahoochee River System as water withdrawal and natural drought have caused flows to diminish. Stream discharge has steadily decreased in the study area by 31% over the last 50 years, and during this time, fish assemblage homogenization has occurred throughout the system. Our primary objective was to assess whether spawning mode and water availability are catalysts for fish assemblage change in tributaries of the Chattahoochee River System. We found that species which prefer or have adapted to low flow conditions (e.g. blacktail shiner Cyprinella venusta, redbreast sunfish Lepomis auritus and blackbanded darter Percina nigrofasciata) are becoming dominant and replacing historically dominant fluvial specialists. When examining short‐term changes between a dry year (2009) and a wet year (2010), some species preferring higher flows such as the highscale shiner Notropis hypsilepis were detected during the wet year, suggesting that the amount of water is important for the presence of these species in upstream reaches where they were historically abundant. Species that can reproduce successfully in low flow conditions are thriving and expanding their native ranges as more habitat becomes suitable. Species that require higher flows for part of their life cycle, however, appear to be declining from upstream areas as water availability continues to decrease, despite temporary recovery from downstream refugia during wet years.     

Aspergillus flavus keratomycosis in a cat treated with topical 1% voriconazole solution

An 8-year-old male castrated Domestic Short-haired cat was examined for a 1-week history of blepharospasm and mucoid ocular discharge OS. Examination revealed ulcerative keratitis with stromal loss, stromal infiltrate, corneal edema, perilimbal vascularization and miosis. Cytology of the cornea revealed multiple dichotomously branching, septate fungal hyphae and severe, predominantly neutrophilic inflammation. PCR of the cytology samples confirmed the presence of Aspergillus flavus while fungal and bacterial cultures were negative. Treatment with topical 1% voriconazole solution was successful in resolving the keratomycosis.     

Avian influenza viruses in wild birds: a moving target

The long-standing evolutionary and ecological relationships between wild birds and influenza A viruses has created a broad pool of viral genetic diversity and a reservoir of potentially transmissible viruses. An understanding of these relationships can help us identify and modify critical control points to reduce transmission of avian influenza viruses into animal and human populations.