A rapid latex agglutination test for gender identification in the Atlantic bluefin tuna,Thunnus thynnus (Linnaeus)

A rapid, one‐step agglutination assay has been developed, based on latex particles sensitized with antibodies against vitellogenin (Vtg), aimed at Atlantic bluefin tuna, Thunnus thynnus (Linnaeus) (ABFT), gender identification. The egg‐yolk precursor protein Vtg was used as a gender marker for the assay as it is a female‐specific protein synthesized during reproductive maturation. The presence of Vtg in the plasma was revealed in 60–120 s through an agglutination reaction by mixing small volumes of ABFT plasma and an anti‐Vtg antibody‐latex suspension on a microscope slide. The effectiveness of the present test was restricted to the months of May and June, concomitant with high circulating Vtg levels. Because of its rapidity and ease of performance in the field, the present gender identification assay could be useful for broodstock management in the aquaculture industry as well as in tagging studies on wild populations.     

Landowners and cat predation across rural-to-urban landscapes

Fluctuations of bird abundances have been attributed to such factors as supplemental feeding, landscape change, and habitat fragmentation. Notably absent from consideration, however, is the role of private landowners and their actions, such as owning free-ranging domestic cats (Felis catus; cats allowed free access to the outdoors). To understand the impacts of cat predation on birds, we surveyed all 1694 private landowners living on three breeding bird survey (BBS) routes (∼120 km) that represent a continuum of rural-to-urban landscapes in Southeastern Michigan, where the majority (>90%) of land is privately owned. Our data indicate that among the 58.5% of landowners that responded, one quarter of them owned outdoor cats. On average a cat depredated between 0.7 and 1.4 birds per week. A total of 23+ species (12.5% of breeding species) were on the list of being killed, including two species of conservation concern (Eastern Bluebirds and Ruby-throated Hummingbirds). Across the three landscapes there were ∼800 to ∼3100 cats, which kill between ∼16,000 and ∼47,000 birds during the breeding season, resulting in a minimum of ∼1 bird killed/km/day. While the number and density (no./ha) of free-ranging cats per landowner differed across the rural to urban landscapes, depredation rates were similar. Landowner participation in bird feeding showed no relationship with the number of free-ranging cats owned. Similarly, selected demographic characteristics of landowners were not significantly related to the number of free-ranging cats owned. Our results, even taken conservatively, indicate that cat predation most likely plays an important role in fluctuations of bird populations and should receive more attention in wildlife conservation and landscape studies.     

A set of highly polymorphic microsatellites useful for kinship and population analysis in turbot (Scophthalmus maximus L.)

The turbot (Scophthalmus maximus) is a flatfish species of great commercial value for aquaculture. In this study we describe the isolation and characterization of 30 novel highly polymorphic microsatellite markers in this species obtained from a genomic library enriched for seven short tandem repeated motifs. Much higher polymorphism (mean number of alleles: 13.37; mean expected heterozygosity: 0.869) and potential for parentage assignment than previously reported for microsatellites in turbot were found after the analysis of 24 wild individuals. Most loci conformed to Hardy–Weinberg expectations, excluding Sma‐USC20 and Sma‐USC28, which showed a high heterozygote deficit probably due to the presence of null alleles. No significant genotypic disequilibrium was observed between any pair of loci, suggesting no close linkage between them. These loci are potentially useful for kinship and population analysis in turbot.     

Ultrasound-guided parasternal injection in dogs: a cadaver study

ObjectivesTo describe the technique of performing an ultrasound-guided distal parasternal intercostal block and to determine the distribution of two volumes of methylene blue dye solution injected in canine cadavers.Study designProspective cadaver study.AnimalsA group of seven canine cadavers weighing 12–34 kg.MethodsThe space between the transversus thoracic and the internal intercostal muscles is a virtual cavity. Ultrasound-guided injections in the distal (parasternal) intercostal space were performed using dye solution at 0.05 mL kg^–1 in each intercostal space from the second to seventh (LV, low volume, six injections per dog) in one hemithorax, and 0.1 mL kg^–1 in the third, fifth and seventh intercostal spaces (HV, high volume, three injections in each dog) on the contralateral side. Anatomical dissection was carried out to describe dye spread characteristics and staining of intercostal nerves.ResultsThe ultrasonographic landmarks for injection were identified in each cadaver. In the LV group the solution was found in every intercostal space (36/36), whereas the HV injection stained six intercostal spaces in two dogs, five in two, and in two dogs the solution was found in four and three spaces, respectively, demonstrating multisegmental distribution. Intrapleural staining was observed after two injections.Conclusions and clinical relevanceUltrasound-guided injection of 0.05 mL kg^–1 at the distal intercostal space resulted in staining of the intercostal nerve in all dogs when performed in every space and may be an appropriate alternative to previously reported techniques. A single injection of 0.1 mL kg^–1 may anaesthetize more than one intercostal nerve, but not consistently. Clinical investigations are warranted to better characterize and to refine this locoregional technique.     

Mycotoxins in corn-based food products consumed in Brazil: an exposure assessment for fumonisins

Samples from 10 different corn-based food products commercially sold in the Federal District of Brazil were analyzed for fumonisins (FB1 and FB2) using HPLC/fluorescence following naphthalene-2,3 dicarboxaldehyde (NDA) derivatization (limit of quantification (LOQ) = 0.020 mg/kg). Samples were also analyzed for aflatoxins (B1, B2, G1, and G2) on a thin-layer chromatrography (TLC) plate under UV light (LOQ of 2 microg/kg). From the 208 samples analyzed, 80.7 and 71.6% had quantifiable levels of FB1 and FB2, respectively. Mean levels of total fumonisins (FB1 + FB2) ranged from 0.127 mg/kg for corn flakes to 2.04 mg/kg for cornmeal ( creme de milho). No FBs were detected in any of the fresh, sweet corn on the cob samples analyzed. Aflatoxins were not detected in any of the 101 samples analyzed. The daily intakes of fumonisins through the consumption of corn-based food products was estimated using consumption data estimated from the 2002/2003 Brazilian Household Budget Survey and the level of fumonisins found in this and other studies conducted in Brazil. In the Federal District, the calculated total daily intake for the total and the consumers-only populations represented, respectively, 9.0 and 159% of the provisional maximum total daily intake (PMTDI) of 2 microg/kg body weight per day. At the national level, the intakes were calculated based on the fumonisin levels found in the Federal District and on published data from studies conducted elsewhere in the country. They represented 24.1 and 355% PMTDI for the total and the consumers-only populations, respectively. The high incidence of fumonisins in some corn-based products and the exposure levels found for specific subpopulations in the present study indicate the need for setting safe regulatory levels for fumonisins in food in Brazil.     

A mixed methodology framework for the assessment of the Voluntary Initiative

The Voluntary Initiative (VI) is a UK programme of measures aimed at reducing the adverse effects of pesticides on the environment. It was initiated in 2001 and will be evaluated during 2006 following a five-year trial period. To assess such a complex scheme, a mixed methodology approach is appropriate, encompassing the assessment of objective deliverables and issues raised by the farming community. A series of focus groups examined the perceptions of the VI by the farming community. Farmer attitudes to the VI appeared to be influenced by the belief that the UK government will, eventually, introduce some form of pesticide tax regardless of the outcome of the VI. Most farmers expressed that they already followed the principles of the VI. It was also evident that the scheme may become more acceptable and better understood as it becomes incorporated into existing farm assurance schemes. This paper proposes a framework for evaluation of the VI that takes into account these perceptions of the VI by the farming community. Based on the analysis of available data, key questions are identified that should be addressed within the final evaluation. These key questions cover the general themes of completion of VI targets and projects, uptake of the VI, environmental impact of the VI and economic implications. To answer the questions, four methods are proposed: a review of literature, a survey of farmers, questions to agencies and environmental modelling.     

Diagnosis of Coxiella burnetii-related abortion in Italian domestic ruminants using single-tube nested PCR

Coxiella burnetii, an obligate intracellular parasite with a worldwide distribution, is the causative agent of acute and chronic Q fever in humans. Although infection is often unapparent in cattle, sheep and goats, there is increasing evidence that C. burnetii infection in these species is associated with abortion and stillbirth. This paper describes the introduction of a single-tube nested PCR protocol for the diagnosis of C. burnetii-related abortion in domestic ruminants in Italy. A total of 514 aborted foetuses from cattle (n = 138) and sheep and goat (n = 376), collected from 301 farms, were analyzed from January 2001 to March 2005. Ninety-seven of 514 (18.9%) animals tested PCR-positive, with 16/138 (11.6%) cattle and 81/376 (21.5%) sheep and goat. Eleven of 102 (10.8%) farms with reproductive disorders in cattle and 37/199 (18.6%) farms with reproductive disorders in sheep and goats were infected with C. burnetii. A greater incidence was observed in three of the seven investigated provinces (p < 0.01), with rates of infected farms of up to 23.8%. Data showed that almost all the C. burnetii-related abortions were recorded between October and April (p < 0.01). These findings suggest that Q fever in humans is largely underestimated in Italy, probably because its occurrence is obscured by flu-like symptoms in acute forms.     

Testing the unifying theory of ozone sensitivity with mature trees of Fagus sylvatica and Picea abies

The broad range in plant responses to chronic O(3) exposure compels a search for integrative, underlying principles. One such approach is the unifying theory proposed by Reich (1987), which combines the O(3) response of contrasting physiognomic classes of plants on the basis of their intrinsic leaf diffusive conductance and, hence, capacity for O(3) uptake. Physiognomic classes differ in the proportional decline in photosynthesis and growth when compared on the basis of cumulative O(3) exposure per unit time, but converge when compared on the basis of O(3) uptake per unit time or cumulative O(3) uptake over the entire lifetime of the leaf. The theory is based on observations on a large number of contrasting plant species, relying primarily on studies of juvenile trees subjected to short-term O(3) exposure. To test the applicability of the unifying theory to mature trees, broadleaf deciduous European beech (Fagus sylvatica L.) and the evergreen conifer Norway spruce (Picea abies (L.) Karst.) in a mature mixed stand were exposed to either ambient air (control) or air with twice the ambient O(3) concentration delivered into the canopy by means of a free-air fumigation system. We accounted for differences in growing season length, leaf longevity and O(3)-related effects on leaf diffusive conductance in determining total O(3) uptake over the lifetime of the leaf. On this basis, Norway spruce needles required 5 years to take up as much O(3) as did beech leaves in one growing season. The core of the unifying theory on O(3) sensitivity was substantiated in relation to O(3) exposure and uptake. However, contrary to the unifying theory, which was formulated on the basis of results with juvenile trees, the O(3) response of mature trees in a natural stand was more complex. The increased complexity was attributed to additional environmental stressors, stress compensation at the whole-tree level, and differential O(3) sensitivities of leaves according to age class and position within the canopy. Contrary to the theory, photosynthesis was no less sensitive to O(3) in Norway spruce than that of beech, and was reduced in the twice-ambient O(3) regime in the first year of exposure.     

Structural basis for double-stranded RNA processing by Dicer

The specialized ribonuclease Dicer initiates RNA interference by cleaving double-stranded RNA (dsRNA) substrates into small fragments about 25 nucleotides in length. In the crystal structure of an intact Dicer enzyme, the PAZ domain, a module that binds the end of dsRNA, is separated from the two catalytic ribonuclease III (RNase III) domains by a flat, positively charged surface. The 65 angstrom distance between the PAZ and RNase III domains matches the length spanned by 25 base pairs of RNA. Thus, Dicer itself is a molecular ruler that recognizes dsRNA and cleaves a specified distance from the helical end.