Comparison of tetrachromic VOF stain to other histochemical staining techniques for characterizing stromal soft and hard tissue components

The components of hard tissues including dentin, enamel, cementum, bone and other calcified deposits, and mature and immature collagen pose problems for identification in routine hematoxylin and eosin (H & E) stained sections. Use of combinations of stains can demonstrate the components of hard tissues and soft tissues distinctly. We assessed the efficacy of the Verde Luz-orange G-acid fuchsin (VOF) stain for differentiating hard and soft connective tissues and compared results with other histochemical staining techniques. Eighty tissue sections comprising developing tooth (30), ossifying fibroma (30) and miscellaneous pathologies (20) expected to contain varying types of calcified tissues were stained with H & E, VOF, and Masson's trichrome (MT). In developing tooth, VOF demonstrated better differentiation of hard tissues, while it was comparable to MT for ossifying fibroma and miscellaneous pathologies. The intensity of staining was greater with VOF than with the other stains studied. VOF stains hard tissue components distinctly and gives good contrast with the surrounding connective tissue. VOF is comparable to MT, but has added advantages including single step staining, rapid and easy procedures, and it distinguishes the maturity of the tissues.     

Sediment phosphate fractionation and interstitial water phosphate concentration in two coastal lagoons (Albuferas de Adra,SE Spain)

The concentrations of o-phosphate and dissolved inorganic carbon were monitored in the interstitial water of the sediment in two highly eutrophic coastal lagoons, Laguna Honda and Laguna Nueva (SE, Spain) from July 2000 to August 2001. Additionally, the organic matter concentration and the P-fractions in the sediment were analysed. Despite their proximity, these two lagoons showed many significant differences (P<0.05). The P-concentration in the interstitial water of the top sediment was higher in the Laguna Honda (up to 1 mg l^–1), especially during summer, when both sites registered their highest values. By contrast, the concentration of organic matter and organic P-fractions were higher in the sediment of the Laguna Nueva. The concentration of the organic matter in the vertical profile of the sediment decreased with depth in the Laguna Nueva (from 17% at 0 cm to 7% at 15 cm, annual mean value) while it increased in the sediment of Laguna Honda (from 9% at 0 cm to 12% at 15 cm, annual mean value). The P-concentration in the interstitial water and the organic matter in the top sediment of Laguna Honda followed a seasonal pattern and, were both correlated with the temperature of the bottom water (r=0.706, P<0.05 and r= -0.929, P<0.01, respectively). The inorg-P fractions comprised 63% of the Tot-phosphate in the sediment of Laguna Honda, whereas org-P fractions (68%) dominated in the sediment of Laguna Nueva. The concentrations of Tot-P in the sestonic material collected at three different depths of the water column averaged 2.2 mg g^–1d.w. in Laguna Honda and 2.5 mg g^–1d.w. in Laguna Nueva. The average atomic C:P ratio of the sestonic material was significantly higher in Laguna Honda than in Laguna Nueva, but the top sediment in Laguna Nueva had a C:P ratio significantly higher than that of the sestonic material (average: 416 and 162, respectively).The different biodegradability of the organic matter in the top sediment probably explained the differences found in sediment composition between these two lagoons. We suggest that the P-reclying from the sediment to the interstitial water was faster in Laguna Honda due to the higher biodegradability of the organic matter. By contrast, the lower biodegradability of the organic matter in the top sediment of Laguna Nueva was probably due to a higher contribution of vascular plants which could explain the organic matter accumulation detected in this lagoon.     

Size and mortality rates of Glossina pallidipes in the semi-arid zone of southwestern Kenya

Seasonal changes in the mean size of tsetse, Glossina pallidipes Austen, as indicated by wing vein length, were monitored during 1983-86 at Nguruman, southwestern Kenya. Changes in size of nulliparous females and wing fray category 1 males were shown to be correlated with the relative humidity 2 months before they were captured. Soil temperature when flies were in the pupal stage had much less effect. Size dependent mortality was demonstrated, with the mean size of flies emerging from pupae significantly less than that of field-caught flies. This mortality must occur at emergence, since there was no evidence of size-dependent mortality once the flies became available to the trap. Size was correlated with density-independent mortality acting on the parent population 2 months previously. It might therefore be possible to use size as an index of the intensity of such mortality. This could be useful when assessing the level of additional mortality required to suppress tsetse populations.     

Nuclear DNA sequences from late Pleistocene megafauna

We report the retrieval and characterization of multi- and single-copy nuclear DNA sequences from Alaskan and Siberian mammoths (Mammuthus primigenius). In addition, a nuclear copy of a mitochondrial gene was recovered. Furthermore, a 13,000-year-old ground sloth and a 33,000- year-old cave bear yielded multicopy nuclear DNA sequences. Thus, multicopy and single-copy genes can be analyzed from Pleistocene faunal remains. The results also show that under some circumstances, nucleotide sequence differences between alleles found within one individual can be distinguished from DNA sequence variation caused by postmortem DNA damage. The nuclear sequences retrieved from the mammoths suggest that mammoths were more similar to Asian elephants than to African elephants.