Monoclonal antibodies identifying chick gonadal cells

1. The aim of the present study was to raise monoclonal antibodies (MAbs) which discriminate gonadal cell types in chickens.

2. A panel of 180 MAbs was generated, from which 14 were chosen for their specificity.

3. These MAbs were characterised immunohistologically on frozen sections of embryonic, newly‐hatched and adult, male and female, left gonads.

4. Three of these MAbs are described: AGC5, AGC7, and AGC13 which recognise respectively germinal epithelial cells, supporting cells and germ cells.

5. These MAbs may be useful in following the developmental pathways in the chicken gonad and analysing the interactive role(s) taking place between gonadal stromal cells and primordial germ cells.     

Variations in the size of the soil biomass

In the surface 5 cm of a clay soil in 2 successive years the microbial biomass, as measured by a fumigation-respiration technique, was constant. The biomasses in two clay soils were approximately ten times greater than that in a silt loam. When straw from a preceding crop was chopped and left on the soil surface the biomass after 8 months was greater by a factor of two than that in soil where the straw had been burnt. Where soil had been kept in grass for 9 yr, the biomass was greater by a factor of three than that in soil of the same kind that had been in arable cultivation for the previous 4 yr. As the fumigation-respiration technique measured only the microbial and microfaunal contribution to the biomass, the total biomass can only be assessed by measuring the root contribution separately; when this was done the total biomass in the grassland was found to be greater by a factor of about six.     

Effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation

OBJECTIVE: To investigate the effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation. SAMPLE POPULATION: Blood samples from 6 Thoroughbreds. PROCEDURE: The degree of fluorescence associated with binding of fluorescein isothiocyanate (FITC)-conjugated anti-human fibrinogen antibody and FITC-annexin V in unactivated and adenosine diphosphate (ADP)-, platelet activating factor (PAF)-, and A23187-activated platelet samples in unfixed and 0.5, 1.0, and 2.0% formaldehyde-fixed samples was assessed by use of flow cytometry. RESULTS: In samples incubated with FITC-anti-human fibrinogen antibody prior to fixation, addition of 2.0% formaldehyde resulted in a 30% increase in total fluorescence in ADP- and PAF-activated samples and a 60% increase in A23187-activated samples. Fixation for 24 hours prior to addition of antibody resulted in reduced fluorescence of samples containing antihuman fibrinogen antibody for all 3 concentrations of formaldehyde in PAF-activated samples. The addition of all 3 concentrations of formaldehyde after incubation with FITC-annexin V resulted in significant increases in fluorescence in unactivated and activated platelet samples. As length of fixation time increased, there was a gradual increase in fluorescence that was significant at 24 hours. CONCLUSIONS: Because fixation with 2.0% formaldehyde results in significant changes in fluorescence in activated platelet samples containing anti-fibrinogen antibody, lower concentrations of formaldehyde should be used to fix equine platelet samples. Formaldehyde-fixed platelet samples should be analyzed within 12 hours of fixation to avoid artifactual increases in fluorescence. Fixation of samples containing FITC-annexin V should be avoided because of significant increases in fluorescence that may interfere with interpretation of results.     

Effects of intravenous administration of formaldehyde on platelet and coagulation variables in healthy horses

OBJECTIVES: To assess safety and determine effects of IV administration of formaldehyde on hemostatic variables in healthy horses. ANIMALS: 7 healthy adult horses. PROCEDURE: Clinical signs and results of CBC, serum biochemical analyses, and coagulation testing including template bleeding time (TBT) and activated clotting time (ACT) were compared in horses given a dose of 0.37% formaldehyde or lactated Ringer's solution (LRS), IV, in a 2-way crossover design. In a subsequent experiment, horses received an infusion of 0.74% formaldehyde or LRS. In another experiment, horses were treated with aspirin to impair platelet responses prior to infusion of formaldehyde or LRS. RESULTS: Significant differences were not detected in any variable measured between horses when given formaldehyde or any other treatment. Infusion of higher doses of formaldehyde resulted in adverse effects including muscle fasciculations, tachycardia, tachypnea, serous ocular and nasal discharge, agitation, and restlessness. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous infusion of formaldehyde at doses that do not induce adverse reactions did not have a detectable effect on measured hemostatic variables in healthy horses.     

Development of a pheromone trap monitoring system for orange wheat blossom midge, Sitodiplosis mosellana, in the UK

Field-trapping experiments with synthetic 2,7-nonadiyl dibutyrate, the female-produced sex pheromone of the orange wheat blossom midge, Sitodiplosis mosellana (Géhin), demonstrated that pheromone traps were highly attractive to males and caught very few non-target organisms. Different formulations of pheromone were tested to identify the optimum release rate and dispenser type for use in pheromone traps in the UK. Key findings were that racemic pheromone was as effective as enantiomerically pure (2S,7R)-2,7-nonadiyl dibutyrate, that release rates higher than 0.5 microg day(-1) were not necessary and that the optimal formulation was a 1 mg pheromone loading in a rubber septum. Pheromone traps gave a reliable indication of peak midge emergence, onset of flight and abundance of midges throughout the season. A strong correlation between maximum trap catch and crop infestation levels was obtained.     

Growth of young cuttlefish, Sepia officinalis (Linnaeus 1758) at the upper end of the biological distribution temperature range

Individual growth rates, feeding rates (%BWd^?1) and food conversions for cuttlefish (S. officinalis) hatchlings and juveniles were determined during this study. A flow‐through system was used. Water temperature reached 30 °C during the hottest part of the day, gradually decreasing to 25 °C during the night; salinity varied between 37 ± 3 ppt and lights were kept on for 14 h day^?1. Hatchlings were placed in separate compartments with a water volume of 1.2 L. Juvenile cuttlefish (from 0.5 to 25 g) were placed in bigger baskets, with a water volume of 5.2 L. Water flow was 120 L h^?1. The biggest cuttlefish used in these experiments (> 25 g) were gathered in groups of five and placed in circular tanks (water volume of 250–300 L). Thus, results obtained in this case are means and not individual data. During the first 10, 20, 30 and 40 days, mean growth rates (of all individuals sampled by age group) decreased consistently (11.8 ± 4.1, 9.8 ± 1.8, 8.1 ± 2.2 and 7.3 ± 0.7%BW^?1 respectively); in similar fashion, mean feeding rates decreased with age group (33.7 ± 13.5, 22.0 ± 7.9, 17.3 ± 3.9 and 16.7%BWd^?1 respectively). Mean food conversions varied between 3.6 and 2.5 between the age groups. When grouping results by weight class, similar patterns occur, as growth and feeding rates decrease consistently as cuttlefish grow bigger. Highest mean growth and feeding rates are obtained by hatchlings (< 0.1 g) with 12.4 ± 4.5 and 35.3 ± 15.1%BWd^?1, respectively, while the lowest growth and feeding rates were recorded for the largest animals, between 15 and 25 g (3.4 ± 1.1 and 10.8 ± 4.1%BWd^?1 respectively). For these weight classes, mean food conversions varied between 2.7 ± 0.9 and 3.8 ± 2.8.     

Dramatic decline in bat species richness in Singapore, with implications for Southeast Asia

Literature searches and recent surveys of the bat fauna of Singapore indicate that of the 24 species of Microchiroptera and six species of Megachiroptera documented for this small equatorial island just 15 and 5, respectively, are still present. These recorded declines in chiropteran species richness almost certainly understate the true losses as extensive land transformation/habitat loss (>95%) and biota loss occurred early in Singapore’s colonial history before comprehensive surveys of bats were made. In an effort to reconstruct the pre-settlement bat fauna, we inferred an upper bound of pre-settlement species richness using a checklist from a well-known bat assemblage in neighbouring Peninsular Malaysia, and a lower bound based on species common to Peninsular Malaysia, Borneo and Sumatra. The Singapore records were compared with these two species list predictions. Based on this analysis, we infer that between 60 and 72 species would have inhabited Singapore before 1819. We also estimate that between 33% (based on the confirmed inventory) and 72% of the species (based on the upper-bound estimate of species richness) are now locally extinct. For Microchiroptera the data suggest that the documented local extinction rate of 38% may project to between 69% and 75%. Forest-dependant bats are particularly affected and comprise a much lower proportion of the bat fauna than is seen in intact forest in Peninsular Malaysia. All hipposiderids and 40% of the documented rhinolophid taxa have been lost and almost half (6) of the surviving microchiropteran species are locally endangered. Projected local extinction rates for Megachiroptera raise the documented value (17%) to about 60%, with most of the survivors being widespread species known to forage in cultivated or secondary forest habitats or to commute long distances between fragmented resources. The dramatic decline in Singapore bat species richness and a concomitant change in chiropteran guild and trophic structure (Microchiroptera vs. Megachiroptera) reflect patterns of diversity change seen elsewhere in the region in response to loss of forest habitat. In Singapore, the decline in diversity (species richness and abundances) for both mega- and microbats may also relate to urbanisation and deliberate or accidental destruction of bats and their roost sites in a land that has one of the highest human population densities on the planet. Although these losses (actual and inferred) represent a microcosm of mainly local extinctions, a wider geographical extrapolation over the 21st century indicates that heavy deforestation in progress in Southeast Asia might be expected to lead to extinction of many bat taxa, with upper-bound estimates of regional species losses exceeding 40% and global extirpation anticipated for at least 23% of Southeast Asia’s bat fauna by 2100.     

A Burkholderia pseudomallei toxin inhibits helicase activity of translation factor eIF4A

The structure of BPSL1549, a protein of unknown function from Burkholderia pseudomallei, reveals a similarity to Escherichia coli cytotoxic necrotizing factor 1. We found that BPSL1549 acted as a potent cytotoxin against eukaryotic cells and was lethal when administered to mice. Expression levels of bpsl1549 correlate with conditions expected to promote or suppress pathogenicity. BPSL1549 promotes deamidation of glutamine-339 of the translation initiation factor eIF4A, abolishing its helicase activity and inhibiting translation. We propose to name BPSL1549 Burkholderia lethal factor 1.