Use of Bullfrog Tadpoles (Rana catesbeiana) to Examine the Mechanisms of Lead Neurotoxicity

Sublethal exposure to lead elicits changes in behavior, particularlylearning. Previously, we had shown that bullfrog tadpoles exposedto 1 mg Pb/liter for 7 days exhibit learning deficits in a discriminateavoidance learning assay. The precise mechanisms involved inthese lead-induced learning deficits are not understood, butCNS monoamine neurotransmitter systems have been implicatedin the learning process. In the present study, we exposed bullfrogtadpoles to 1.7± 0.2 mg Pb/liter for 7 days and thencompared concentrations of neurotransmitters from whole brainsamples with those of controls. Serotonin or 5-hydroxytryptamine(5-HT) was significantly decreased in the lead exposed groupwhile the serotonin metabolite, 5-hydroxyindoleacetic acid,(5-HIAA) was similar to controls. No changes were observed inthe catecholamines epinephrine and norepinephijine followinglead exposure. The ratio of 5-HIAA/5-HT was significantly increasedin lead exposed animals as a result of the decrease in 5-HT,suggesting a decrease in 5-HT biosynthesis rather than an increasein 5-HT metabolism. These findings are the first to suggestthat lead exposure in bullfrog tadpoles affects the monoamineneurotransmitters that are implicated in the learning process.The results of the present study, in conjunction with previousevidence of learning deficits following lead exposure, offerthe possibility of correlating lead exposure with learning deficitsand alterations in CNS neurotransmitters in bullfrog tadpoles.The use of this tadpole model shows promise as a means to examineand understand the mechanisms involved in lead neurotoxicity.     

Landscape structure influences tree density patterns in fragmented woodlands in semi‐arid eastern Australia

Landscape and local‐scale influences are important drivers of plant community structure. However, their relative contribution and the degree to which they interact remain unclear. We quantified the extent to which landscape structure, within‐patch habitat and their confounding effects determine post‐clearing tree densities and composition in agricultural landscapes in eastern subtropical Australia. Landscape structure (incorporating habitat fragmentation and loss) and within‐patch (site) features were quantified for 60 remnant patches of Eucalyptus populnea (Myrtaceae) woodland. Tree density and species for three ecological maturity classes (regeneration, early maturity, late maturity) and local site features were assessed in one 100 × 10 m plot per patch. All but one landscape characteristic was determined within a 1.3‐km radius of plots; Euclidean nearest neighbour distance was measured inside a 5‐km radius. Variation in tree density and composition for each maturity class was partitioned into independent landscape, independent site and joint effects of landscape and site features using redundancy analysis. Independent site effects explained more variation in regeneration density and composition than pure landscape effects; significant predictors were the proportion of early and late maturity trees at a site, rainfall and the associated interaction. Conversely, landscape structure explained greater variation in early and late maturity tree density and composition than site predictors. Area of remnant native vegetation within a landscape and patch characteristics (area, shape, edge contrast) were significant predictors of early maturity tree density. However, 31% of the explained variation in early mature tree differences represented confounding influences of landscape and local variables. We suggest that within‐patch characteristics are important in influencing semi‐arid woodland tree regeneration. However, independent and confounding effects of landscape structure resulting from previous vegetation clearing may have exerted a greater historical influence on older cohorts and should be accounted for when examining woodland dynamics across a broader range of environments.     

Field resistance in cowpea, Vigna unguiculata to the cowpea moth, Cydia ptychora

Field evaluation of 26 selected cowpea cultivars showed that several of them had good levels of resistance to Cydia ptychora; TVu 946, Adzuki, ‘Vita 5’ and Igbirra were the most resistant. Percentage seed damaged by the larvae was positively correlated with days to 50% flowering (r= 0.39*), days to 50% pod ripening (r= 0.48*) and weight of 100 seeds (r= 0.56**). All four cultivars were early maturing, had semi-determinate or determinate plant types and carried their pods above the leaf canopy. No single morphological character could be related to the degree of seed damage but some association with phenological characteristics was indicated. Earliness, determinacy and uniform pod maturity were important plant characters associated with the expression of field resistance. Even if these characters are not causative, they provide a rational basis for screening of a large germplasm to identify or synthesise materials with greater resistance to this moth.     

High‐throughput species identification: from DNA isolation to bioinformatics

Ichthyoplankton collections provide a valuable means to study fish life histories. However, these collections are greatly underutilized, as larval fishes are frequently not identified to species due to their small size and limited morphological development. Currently, there is an effort underway to make species identification more readily available across a broad range of taxa through the sequencing of a standard gene. This effort requires the development of new methodologies to both rapidly produce and analyse large numbers of sequences. The methodology presented in this paper addresses these issues with a focus on the larvae of large pelagic fish species. All steps of the methodology are targeted towards high‐throughput identification using small amounts of tissue. To accomplish this, DNA isolation was automated on a liquid‐handling robot using magnetic bead technologies. Polymerase chain reaction and a unidirectional sequencing reaction followed standard protocols with all template cleanup and transferring also automated. Manual pipetting was thus reduced to a minimum. A character‐based bioinformatics program was developed to handle the large sequence output. This program incorporates base‐call quality scores in two types of sample to voucher sequence comparisons and provides suggested identifications and sequence information in an easily interpreted spreadsheet format. This technique when applied to tuna and billfish larvae collected in the Straits of Florida had an 89% success rate. A single species (Thunnus atlanticus) was found to dominate the catch of tuna larvae, while billfish larvae were more evenly divided between two species (Makaira nigricans and Istiophorus platypterus).     

PCR primers for polymorphic microsatellite loci in the plant‐ant Azteca ulei cordiae (Formicidae: Dolichoderinae)

Twelve microsatellite loci were isolated from the Peruvian tropical plant‐ant, Azteca ulei cordiae (Hymenoptera: Dolichoderinae). High levels of within‐population variation were observed at most loci, with number of alleles ranging from four to 18, and heterozygosity from 0.118 to 1 per population sample. Cross‐species amplification of these loci was also successfully tested in several other species of the same ant genus Azteca.     

Comparison of 5′-Nucleotidase Activities Among Cultured Murine Melanoma Cell Lines

The activity of 5′-nucleotidase and ouabain-sensitive Na/K ATPase was determined in seven different mouse melanoma cell lines. Ouabain-sensitive Na/K ATPase activity was found in NP40-treated cell homogenates of all cell lines. However, 5′-nucleotidase activity was found in only one mouse melanoma cell line—JB/RH. The absence of expression of 5′-nucleotidase activity in the other six cell lines is not associated with pigmentation in melanoma cells, nor is the gene switched off in all transformed melanocytes of C57BL/6 origin.