Clearing and dissecting insects for internal skeletal morphological research with particular reference to bees

ABSTRACT A detailed protocol for chemical clearing of bee specimens is presented. Dry specimens as well as those preserved in liquid media can be cleared using this protocol. The procedure consists of a combined use of alkaline solution (KOH or NaOH) and hydrogen peroxide (H2O2), followed by the boiling of the cleared specimens in 60–70% EtOH. Clearing is particularly useful for internal skeletal morphological research. This procedure allows for efficient study of internal projections of the exoskeleton (e.g., apodemes, furcae, phragmata, tentoria, internal ridges and sulci), but this process makes external features of the integument, as some sutures and sulci, readily available for observation as well. Upon completion of the chemical clearing process the specimens can be stored in glycerin. This procedure was developed and evaluated for the preparation of bees and other Apoidea, but modifications for use with other insect taxa should be straightforward after some experimentation on variations of timing of steps, concentration of solutions, temperatures, and the necessity of a given step. Comments on the long-term storage, morphological examination, and photodocumentation of cleared specimens are also provided.

Variability of apparently homogeneous soilscapes in São Paulo state, Brazil: I. spatial analysis

The spatial variability of strongly weathered soils under sugarcane and soybean/wheat rotation was quantitatively assessed on 33 fields in two regions in São Paulo State, Brazil: Araras (15 fields with sugarcane) and Assis (11 fields with sugarcane and seven fields with soybean/wheat rotation). Statistical methods used were: nested analysis of variance (for 11 fields), semivariance analysis and analysis of variance within and between fields. Spatial levels from 50 m to several km were analyzed. Results are discussed with reference to a previously published study carried out in the surroundings of Passo Fundo (RS). Similar variability patterns were found for clay content, organic C content and cation exchange capacity. The fields studied are quite homogeneous with respect to these relatively stable soil characteristics. Spatial variability of other characteristics (resin extractable P, pH, base- and Al-saturation and also soil colour), varies with region and, or land use management. Soil management for sugarcane seems to have induced modifications to greater depths than for soybean/wheat rotation. Surface layers of soils under soybean/wheat present relatively little variation, apparently as a result of very intensive soil management. The major part of within-field variation occurs at short distances (< 50 m) in all study areas. Hence, little extra information would be gained by increasing sampling density from, say, 1/km² to 1/50 m². For many purposes, the soils in the study regions can be mapped with the same observation density, but residual variance will not be the same in all areas. Bulk sampling may help to reveal spatial patterns between 50 and 1.000 m.