Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant Factor C assay

Measurement of environmental endotoxin exposures is complicated by variability encountered using current biological assay methods arising in part from lot-to-lot variability of the Limulus-amebocyte lysate (LAL) reagents. Therefore, we investigated the lot-to-lot repeatability of commercially available recombinant Factor C (rFC) kits as an alternative to LAL. Specifically, we compared endotoxin estimates obtained from rFC assay of twenty indoor dust samples, using four different extraction and assay media, to endotoxin estimates previously obtained by Limulus amebocyte lysate (LAL) assay and amounts of 3-hydroxy fatty acids (3-OHFA) in lipopolysaccharide (LPS) using gas-chromatography mass spectroscopy (GC-MS). We found that lot-to-lot variability of the rFC assay kits does not significantly alter endotoxin estimates in house dust samples when performed using three of the four assay media tested and that choice of assay media significantly altered endotoxin estimates obtained by rFC assay of house dust samples. Our findings demonstrate lot-to-lot reproducibility of rFC assay of environmental samples and suggest that use of rFC assay performed with Tris buffer or water as the extraction and assay medium for measurement of endotoxin in dust samples may be a suitable choice for developing a standardized methodology.     

Photocatalytic degradation of Chicago Sky Blue 6B and Benzopurpurin 4B using titanium dioxide thin film

Aqueous solutions of azo dyes undergo degradation to form harmless intermediates and colorless products following irradiation by visible light in the presence of titanium dioxide thin films. The dyes that were studied in this work are： Chicago Sky Blue 6B and Benzopurpurin 4B. Results obtained indicated that complete mineralization of the dyes took place under the experimental conditions. There was an increase in conductivity after the complete mineralization experiments possibly indicating the formation of ions such as NO3^- and SO4^2-. Chemical oxygen demand（COD） measurements show a decrease in organic matter for both dyes following complete degradation. The effect of how changing experimental conditions such as pH, temperature and starting concentrations of dyes affected the rate of dye degradation was measured. There was an increase in the rate of disappearance of the dye color at lower pH. High concentrations of dye solutions reauired Iona dearadation time.     

A dynamic transmission model of eastern equine encephalitis virus

Eastern equine encephalitis virus (EEEV) is one of several arthropod-borne viruses (arboviruses) endemic to the United States. Interactions between arthropod (mosquito) vectors and avian amplification host populations play a significant role in the dynamics of arboviral transmission. Recent data have suggested the hypothesis that an increased rate of successful feeding on young-of-the-year (YOY) birds might play a role in the dynamics of EEEV transmission. To test this hypothesis, we developed a model to explore the effect of the interactions of the vectors and avian host populations on EEEV transmission. Sensitivity analyses conducted using this model revealed eleven parameters that were capable of disproportionately affecting the predicted level of EEEV infection in the vertebrate reservoir and vector populations. Of these, four parameters were related to the interaction of the vector with young-of-the-year birds. Furthermore, adult birds could not substitute for young-of-the-year in initiating and maintaining a predicted enzootic outbreak of EEEV. Taken together, the model predicted that young-of-the-year birds play a key role in establishing and maintaining enzootic outbreaks of EEEV.     

Energy investment in the annual reproduction cycle of female red porgy, <Emphasis Type="Italic">Pagrus pagrus</Emphasis> (L.)

Seasonal cycle in energy content of gonad, perivisceral fat, liver, and muscle was used to estimate the energy cost of spawning of female red porgy (Pagrus pagrus) from Buenos Aires coasts, Argentina. Spawning was observed primarily between November and January, although a low-intensity spawning was also observed at the beginning of this period. The energy required for gonad development of female red porgy (the difference between the maximum and minimum energy content of ovaries along the year) was 2,652 J g⁻¹ dry weight or 558 kJ. Female red porgy expend 22.8% of their total body energy (3,481 kJ) during the spawning period. Of this energy, relative contributions from different sources were as follows: ovaries 62.5% (496 kJ), perivisceral fat 17.8% (142 kJ), liver 10.3% (82 kJ), and muscles 9.4% (75 kJ). Female spawners lost about 7.2% (119 g) of their total body weight (1,654 g) in the spawning period. From the 7.2% of the total weight lost, female gonad represented a relative reduction of 73.8% (87.6 g), the perivisceral fat declined by 5.7% (6.8 g), the liver declined by 10.6% (12.5 g), and muscle tissue exhibited a 10% loss in total weight (11.9 g). Reproductive effort represented 7.6% of food energy, while resting metabolism accounted for almost 38% of food energy. Despite the fact that red porgy females feed throughout the entire spawning period, it is shown that between pre-spawning (September, water temperature 15°C) and mid-spawning (December, water temperature 19°C) they lost 45 and 38% of their muscle and liver lipids, respectively, and 89% of the perivisceral fat associated with the gut and body cavity. Red porgy used both their energy reserves and food energy during ovarian maturation. This asynchronous species have a strategy of a lower energy allocation and a higher dependence of food during the spawning time (late-spring and summer) for successful reproduction.     

Role of sex and breeding status in grooming and total tick load of impala

The lesser mouse lemur (Microcebus murinus) is a prosimian primate which presents evidence of sex ratio bias of offspring that agrees with the direction of bias predicted by the local resource competition model for facultative sex ratio adjustment. That is, females overproduced sons when grouped prior to mating, whereas isolated females exhibited the opposite tendency. In this solitary species, social communication relies heavily on urinary chemical signals. To test the hypothesis that sex biases induced by social factors may be linked to urinary cues, isolated females were exposed (n = 76) or not (control group, n = 16) to urinary cues from other reproductively active females from the beginning of the breeding season (induced by long photoperiod) until oestrus. During that period, females were either continuously (n = 17) or partially (n = 59) exposed to chemosignal stimulation. Females in oestrus were placed in contact with a breeding male and subsequently isolated until they gave birth. All females entered oestrus but the timing of oestrus was significantly delayed by 1 week in urine-exposed females. A general depressive effect of long-term urine exposure on fecundity was demonstrated, involving fewer impregnations, higher abortion frequency and smaller litter sizes. Among females giving birth (n = 55) to a total of 129 young, a significant positive correlation was found between sex ratio bias towards males and the duration of urine exposure. However, the shift in sex ratio at birth depended on the duration of urine stimulation during a sensitive period extending from the beginning of the long photoperiod until the beginning of the follicular phase. In the absence of urinary cues during the sensitive period, females significantly overproduced daughters (32% males of 53 newborn). As urine exposure increased during the sensitive phase, the proportion of males in litters increased from 54% males (n = 50) in partially urine-exposed females to a significant bias towards males (69.2% of 26 newborn) in totally exposed females. The biased sex ratio in response to chemical cues did not show consistent relationships with maternal body weight, parity or litter size. Although the intrinsic mechanisms involved in sex-biased conceptions are not known, chemical cues could interact with the photoperiodic control of gonadotropin secretions. Received: 14 January 1995/Accepted after revision: 26 November 1995     

A SPE-HPLC-MS/MS method for the simultaneous determination of prioritised pharmaceuticals and EDCs with high environmental risk potential in freshwater

This work describes the development, optimisation and validation of an analytical method for the rapid determination of 17 priority pharmaceutical compounds and endocrine disrupting chemicals (EDCs). Rather than studying compounds from the same therapeutic class, the analyses aimed to determine target compounds with the highest risk potential (with particular regard to Scotland), providing a tool for further monitoring in different water matrices. Prioritisation was based on a systematic environmental risk assessment approach, using consumption data; wastewater treatment removal efficiency; environmental occurrence; toxicological effects; and pre-existing regulatory indicators. This process highlighted 17 compounds across various therapeutic classes, which were then quantified, at environmentally relevant concentrations, by a single analytical methodology. Analytical determination was achieved using a single-step solid phase extraction (SPE) procedure followed by high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The fully optimised method performed well for the majority of target compounds, with recoveries >71% for 15 of 17 analytes. The limits of quantification for most target analytes (14 of 17) ranged from 0.07 ng/L to 1.88 ng/L in river waters. The utility of this method was then demonstrated using real water samples associated with a rural hospital/setting. Eight compounds were targeted and detected, with the highest levels found for the analgesic, paracetamol (at up to 105,910 ng/L in the hospital discharge). This method offers a robust tool to monitor high priority pharmaceutical and EDC levels in various aqueous sample matrices.     

Brittlestars,Biomarkers and Beryl: Assessing the Toxicity of Oil-Based Drill Cuttings Using Laboratory,Mesocosm and Field Studies

Effects of drill cuttings on burrowing brittlestars (Amphiura chiajei and A. filiformls) were compared at different levels of response, from acute laboratory experiments to chronic field studies. Acute toxicity tests showed that drill cuttings containing oil-based muds had a very low toxicity (LC₅₀ 52,800 ppm total hydrocarbons in test sediment) but sub-lethal effects were detected at much lower levels using a variety of biomarkers and assays. Chronic sub-lethal effects were detected around the Beryl oil platform in the North Sea where the levels of oil in the sediment were very low (<3 ppm total hydrocarbons in sediment) and brittlestars were excluded from areas nearer the platform with higher sediment oil content. These results suggest that acute toxicity tests are a poor predictor of chronic response in these animals. the relationship between toxicity at different levels of response and the predictive power of laboratory studies is discussed.     

Intracellular bacteria associated with the ascidian<Emphasis Type="Italic"> Ecteinascidia turbinata</Emphasis>: phylogenetic and in situ hybridisation analysis

The ascidian Ecteinascidia turbinata (Herdman) is a colonial sea squirt found in the Caribbean and Mediterranean Seas. In the present study, the bacterial complement of E. turbinata has been assessed by 16S rRNA gene analysis and the most commonly occurring strains identified by restriction fragment length polymorphism. Three strains were found to predominate using this approach, with one representing >50% of clones from both larval and adult material. The 16S rRNA gene sequence of the most commonly occurring strain did not match with any known bacterial sequences and could only be assigned to the γ-proteobacteria subdivision. The two other frequently occurring strains were assigned to the Mollicutes. In situ hybridisation analysis with eubacterial probes to 16S rRNA revealed the presence of apparently endosymbiotic bacteria in adult and larval tissue, and electron microscopy confirmed the presence of putative bacteriocytes in the larval tissue. The presence of the same bacteria in the brooded larvae suggested that they were vertically transmitted from parent to offspring. Further hybridisation using a novel probe designed to be specific to the 16S rRNA sequence of the dominant strain, highlighted the same cell types as that revealed by the eubacterial probe. The results suggest that the bacteria represent a novel strain, denoted "Candidatus Endoecteinascidia frumentensis", and that they may have an important role in the biology of E. turbinata.     

SOME SIMPLE MODELS FOR DISCRETE VARIATE TIME SERIES1

ABSTRACT: Simple models are presented for use in the modeling and generation of sequences of dependent discrete random variables. The models are essentially Markov Chains, but are structurally autoregressions, and so depend on only a few parameters. The marginal distribution is an intrinsic component in the specification of each model, and the Poisson, Geometric, Negative Binomial and Binomial distributions are considered. Details are also given for the introduction of time-dependence into the means of the sequences so that seaonality can be treated simply.     

Multiple paternity in meadow voles (Microtus pennsylvanicus): investigating the role of the female

Multiple paternity in single litters conceived in the wild was recently demonstrated in meadow voles (Microtuspennsylvanicus). In this study, we used an experimental approach (males tethered and females allowed to mate freely with one or several males) to investigate the role of female meadow voles in multiple paternity. We found that among 29 (of 39) females that copulated during our experiment, 79.3% chose to mate with more than one male. Female behavior in meadow voles thus clearly promotes multiple paternity and their role is an active one. Some of the hypotheses explaining promiscuity in meadow voles should be reconsidered in light of this result. We do not know the primary determinant of female mate choice, but male body mass played a secondary role in driving female preferences. The partial dependence between male body mass and female choice, coupled with the active role played by females, indicates that intersexual selection has the potential for reinforcing the effects of intrasexual selection (male-male dominance relationships) in this species. Finally, we demonstrate that the time period over which tests are conducted is an important part of the design of experiments aimed at understanding the role of females in multiple paternity. Received: 14 April 1998 / Accepted after revision: 12 September 1998