Evidence that active transmission of porcine cysticercosis occurs in Venezuela

There is a paucity of quantitative data on the status of porcine cysticercosis in Venezuela, information which is essential for understanding the level of disease transmission. This study was, therefore, conducted in a typical small rural community in Yaracuy State, Venezuela, where previous cases of human Taenia solium taeniasis/cysticercosis had been reported and where the free-ranging pig management practices and the lack of rudimentary sanitary facilities indicated an obvious risk for transmission of the disease. Serum samples from 52 village pigs were screened by enzyme-linked immunosorbent assays for anti-cysticercal antibodies (Ab-ELISA), using T. solium cyst fluid as the antigen and the HP10, monoclonal antibody-based, antigen trapping ELISA for parasite antigen (HP10 Ag-ELISA). Significantly, a high proportion of the animals (65.4% for the Ab-ELISA and 42.3% for the HP10 Ag-ELISA) were sero-positive. Five of the pigs, which were selected on that basis of positive tongue palpation, were killed for autopsy, and large numbers of viable cysticerci were found in the carcases. This unequivocal documentation of porcine cysticercosis in Venezuelan pigs presents clear evidence that T. solium is actively transmitted in Venezuela. Further detailed studies and implementation of appropriate control measures are therefore indicated.     

Methods for Determining Relative Average Number of Channels per Maize Starch Granule and Digestion of Raw Granules of Mutant Maize Cultivars by Amyloglucosidase

Six methods were investigated for determining the relative average numbers of channels per granule in populations of starch granules. The first method involved digestion of raw starch granules with amyloglucosidase. There were differences in the rates of digestion of starch of the same mutation in two different inbred‐line backgrounds, but we concluded that a better method was needed. Three methods involved measuring either all channel proteins or one specific protein. Extraction of granules using 70% 2‐propanol containing 1% 2‐mercaptoethanol (ME) was successful in removing the majority of external surface proteins (<31 kDa). Further extraction of starch granules using 0.1% SDS containing 0.2% ME, pH 8, removed channel proteins without extracting matrix proteins. By measuring SDS‐PAGE band intensities of protein clusters at 31–97.4 kDa or a marker protein at ≈42 kDa, methods to quantify relative average degrees of channelization (RADC) in different maize backgrounds were developed. Using either method, Oh43 had the highest RADC and B73 had the lowest RADC among the inbred lines examined. Another method involves extraction of total proteins using 2% SDS containing 0.2% ME from granules with the majority of surface proteins removed. The extract is subjected to SDS‐PAGE and Western blotting, followed by location and quantification of the actin band using antibodies to maize pollen actin. These four methods give somewhat different results for six inbred lines of corn with a relatively narrow range of RADC. In an attempt to relate them to the actual number of observed pores (channel openings to the surface), granules were treated with α‐amylase to enlarge the pores, with and without prior treatment with a protease. We concluded that this treatment revealed something about the nature of the granules and the action of α‐amylase on them, but it was not an indication of the number of channels present. The outcome of this research provides evidence that the number of channels in corn starch granules varies with the genetic makeup of the parent plant. We concluded that measuring granule actin is the preferred method.     

Adventitious root cultures of Castilleja tenuiflora Benth. as a source of phenylethanoid glycosides

Castilleja tenuiflora is a highly valued medicinal plant that grows in pine-oak woods in Mexico. In this study, we identified for the first time verbascoside and isoverbascoside as the major phenylethanoid glycosides (PhGs) in C. tenuiflora. These compounds have proven biological activities, including anti-inflammatory, antioxidant, and cytotoxic activities, which may be related to the traditional uses of this plant. We developed a reverse-phase high-performance liquid chromatography (RP-HPLC) procedure to analyze PhGs, and determined their concentrations in various different tissues of wild plants. Verbascoside accumulated mainly in roots and inflorescences (9.23 and 7.88 mg g⁻¹ dry biomass, respectively), while isoverbascoside accumulated mainly in the roots (7.13 mg g⁻¹ dry biomass). To provide an alternative source of material for production of bioactive compounds, we established in vitro adventitious root cultures in which roots were grown in B5 medium containing either 10 μM indole 3-acetic acid (IAA) or 10 μM α-naphthaleneacetic acid (NAA). The greatest dry biomass yield (30 g L⁻¹) was achieved at 30 days after transfer of roots into IAA-containing medium. The highest specific yields of PhGs were also obtained using this auxin; the maximum level of verbascoside was 14.62 mg g⁻¹ dry root biomass (438.6 mg L⁻¹) at 30 days after root transfer, and the maximum yield of isoverbascoside was 37.32 mg g⁻¹ dry root biomass (522.48 mg L⁻¹) at 23 days after root transfer. Adventitious root cultures of C. tenuiflora are a promising system for further studies on scale-up and phenylethanoid glycosides biosynthesis.