Morphology versus molecules: sexing Red-winged Blackbird nestlings

ABSTRACT.   Past studies of offspring sex ratios in birds have often relied on sexually size dimorphic species where nestling sex could be determined based on weight at a given age. DNA-based sexing techniques allow us to assess the accuracy of those techniques and to refine them for use when costs or convenience make DNA methods impractical. Using nestling Red-winged Blackbirds ( Agelaius phoeniceus ) whose sex was determined using DNA, we compared sex ratios obtained using different morphological criteria. Conservative criteria from previous studies were completely accurate, but allowed sexing of few nestlings younger than 8 d old, and were more successful for sexing males than females. A new method was developed that allowed accurate sexing of nestlings beginning at day 6 posthatching and was less biased relative to known sex ratios. Using 11 years of data, the conservative method left an average of 55% of nestlings and 36% of fledglings unsexed, compared to 31% and 9% using the new method. Furthermore, the male bias in sex ratio estimates using the conservative method was greater, both absolutely and relative to estimates based on the new method, when the proportion of unsexed nestlings (because they were not weighed when older) was higher. Thus, estimates of population sex ratios will be more accurate as the number of nestlings measured on day 8 or older increases. However, if some nestlings that were not weighed past day 7 fledge, the new method allows more of those individuals to be sexed than the conservative method, and the population sex ratio estimate should be more reliable. Although our approach should apply to other sexually dimorphic species, the criteria used must be developed based on such species-specific attributes as growth patterns and degree of hatching asynchrony.     

Regional genetic variation in the major sperm protein genes of Onchocerca volvulus and Mansonella ozzardi (Nematoda: Filarioidea)

Onchocerca volvulus and Mansonella ozzardi are two human filarial parasites present in South and Central America. In the Brazilian Amazonia they are found in sympatry, and the lack of clear morphological diagnostic characters in the microfilariae hinders their identification. The major sperm protein (MSP) gene of both species has been sequenced and characterised to determine its potential as a molecular diagnostic character. The length of the MSP gene is different in each species, and this could be used to detect and differentiate them by running the polymerase chain reaction (PCR) product in an agarose gel. Two major gene groups were identified in O. volvulus with a genetic distance of 6% between them. In M. ozzardi only one major group of genes was observed. The high similarity between the protein amino acid sequence of both filarial species confirms that the MSP has been highly conserved through nematode evolution.     

Phytochemical data parallel morpho-colorimetric variation in Polygala flavescens DC.

Abstract

Phytochemical data, integrated with other sources of information, represent a valuable tool helping to solve different kinds of taxonomic problems in plant systematics. In the present study, a comparative investigation, in order to clarify the systematic relationships of the three subspecies currently recognized within the Italian endemic Polygala flavescens, was carried out. Preliminarily, a morphometric and colorimetric analysis, in order to test the degree of morphological distinctiveness among the taxa, was performed. Then, a phytochemical analysis based both on volatile and non-volatile compounds was obtained. Concerning the morpho-colorimetric analysis, our results confirm most of the characters as useful to discriminate the three subspecies. In addition, some volatile and non-volatile compounds are good taxonomic markers. Morpho-colorimetric variation is clearly paralleled by phytochemical results, confirming the value of this kind of data to infer relationships in plant systematics. Based on these results, we support a taxonomic treatment at subspecific level for the involved taxa. Finally, based on the most significant morphological characters, a revision of herbarium specimens allowed to redefine the distribution pattern of the three subspecies. Accordingly, the range of P. flavescens subsp. maremmana is limited to Mt. Argentario (southern Tuscany) only. A key is also reported for the identification of the three subspecies.     

Species identification in meat by using PCR-generated satellite probes

A convenient DNA-based identification system is described for testing the species origin of meat samples. Probes are generated by PCR with primers binding to species-specific satellite DNA and hybridized to DNA purified from meat. This method is more robust and versatile than methods based on oligonucleotide hybridization. With the exception of a slight cross-reaction of mutton and beef, each probe only recognized the species from which it was derived. Purifying the DNA with a DNA-binding resin improved the sensitivity. Admixtures of 0.1–0.5% can be detected in raw meat and 0.5–5% in autoclaved meat samples. The method can be adapted to detect any eukaryotic species for which species-specific DNA sequences are available. This method has proven its value in the routine inspection of meat samples by revealing more cases of deliberate or accidental species substitution and admixture than conventional techniques. Received 12 January 1998/ Accepted in revised form 15 June 1998     

Eighteen polymorphic microsatellite markers for the highly endangered Spanish imperial eagle (Aquila adalberti) and related species

Here we describe the development of 18 polymorphic microsatellite markers for the endangered Spanish imperial eagle (Aquila adalberti). Microsatellites were tested in five other raptor species. These markers were revealed as good molecular tools for genetic population studies, individual identification and parentage assessment in Spanish imperial eagle and closely related species.     

An ecological approach to restructuring school biology

As a follow-up to the human-centred and cybernetic structuring principles described by Kattman and Schaefer in J. Biol. Educ. 10 (3), an ecological structuring principle is discussed. This principle takes the dynamic and reciprocal relationships between man and his environment as the basis of his existence. The reciprocal relationships are characterized as the biological capacity for self-preservation, reproduction, and adaptation. Several examples from a teaching syllabus are given to illustrate this approach.     

Microarray-based genetic identification of beneficial organisms as a new tool for quality control of laboratory cultures

The use of beneficial organisms to help control pests and pathogens in field and greenhouse crops is constantly increasing. Insects and mites are commonly used as beneficial organisms and, nowadays, rearing companies have to produce them in large quantities. Because of the peculiarities of laboratory culture conditions, the quality of lab-reared organisms generally degrades over time. To maintain high fitness levels, cultures are refreshed with field specimens at regular intervals. However, this bears the risk of contaminating laboratory cultures with species or strains other than the intended natural enemy. To ensure that the correct species is produced and also to facilitate surveys after field release, we have developed a diagnostic microarray for identification of beneficial species. Probes have been designed from the different haplotypes of a fragment of the mitochondrial cytochrome oxidase I (COI) gene of each species. Hybridization of labeled PCR amplicons of COI on the microarray chip allows precise identification of 28 economically relevant arthropod species.