Evaluation of steroidogenic capacity after follicle stimulating hormone stimulation in bovine granulosa cells of Revalor 200? implanted heifers

Background

Heifers not used as breeding stock are often implanted with steroids to increase growth efficiency thereby altering hormone profiles and potentially changing the environment in which ovarian follicles develop. Because bovine granulosa cell culture is a commonly used technique and often bovine ovaries are collected from abattoirs with no record of implant status, the objective of this study was to determine if the presence of an implant during bovine granulosa cell development impacts follicle stimulating hormone-regulated steroidogenic enzyme expression. Paired ovaries were collected from 16 feedlot heifers subjected to 1 of 3 treatments: non-implanted (n = 5), Revalor 200 for 28 d (n = 5), or Revalor 200 for 84 d (n = 6). Small follicle (1 to 5 mm) granulosa cells were isolated from each pair and incubated with phosphate buffered saline (n = 16) or 100 ng/mL follicle stimulating hormone (n = 16) for 24 h.

Results

Granulosa cells of implanted heifers treated with follicle stimulating hormone produced medium concentrations of progesterone similar (P = 0.22) to non-implanted heifers, while medium estradiol concentrations were increased (P < 0.10) at 28 and 84 d compared to non-implanted heifers indicating efficacy of treatment. Additionally, real-time PCR analysis in response to follicle stimulating hormone treatment demonstrated a decrease in steroidogenic acute regulatory protein (P = 0.05) mRNA expression in heifers implanted for 84 d and an increase in P450 side chain cleavage mRNA in granulosa cells of heifers implanted for 28 (P < 0.10) or 84 d (P < 0.05) compared to non-implanted females. However, no difference in expression of 3-beta-hydroxysteroid dehydrogenase (P = 0.57) and aromatase (P = 0.23) were demonstrated in implanted or non-implanted heifers.

Conclusions

These results indicate follicles which develop in the presence of high concentrations of androgenic and estrogenic steroids via an implant tend to demonstrate an altered capacity to respond to follicle stimulating hormone stimulation. Thus, efforts should be made to avoid the use of implanted heifers to study steroidogenesis in small follicle granulosa cell culture systems.     

Historical DNA as a tool to genetically characterize the Mediterranean sand tiger shark (Carcharias taurus,Lamniformes: Odontaspididae): A species probably disappeared from this basin

1. The sand tiger shark (Carcharias taurus) is a coastal species distributed in temperate and sub‐tropical waters, classified as ‘Vulnerable’ at global level and ‘Critically endangered’ in eastern Australia, south‐western Atlantic Ocean, and Mediterranean Sea. Six populations (north‐western Atlantic, Brazil, South Africa, Japan, eastern Australia, and western Australia) with low genetic diversity and limited gene flow were identified worldwide, but genetic information for many other geographic areas are still missing. Specifically, this species is listed in several reports as part of the Mediterranean fauna, even if there has been a lack of catches and sightings in recent years in this basin. To clarify the origin of C. taurus individuals caught in the past in the Mediterranean Sea, historical samples were genetically analysed.
2. Nine samples with certain Mediterranean origin were collected from different European museums. DNA was extracted and ~600 bp of the mitochondrial DNA control region were amplified using eight overlapping species‐specific primer pairs. Sequences obtained were aligned with all the haplotypes globally known so far.
3. Genetic analysis revealed the misidentification of one museum specimen. Among the remaining Mediterranean historical samples, three different haplotypes were recovered. Two of them previously observed only in South Africa and one described in both South African and Brazilian populations.
4. Results suggest a genetic relationship between Mediterranean sand tiger sharks and those from the western Indian Ocean. According to previous studies, we hypothesized that, during the Pleistocene, the cold Benguela upwelling barrier was temporarily reduced allowing the passage of C. taurus individuals from the Indian to Atlantic Ocean. After the restoration of this phylogeographic barrier some individuals were trapped in the Atlantic Ocean and probably migrated northward colonizing the western African coasts and the Mediterranean Sea.

     

Disentangling effects of habitat diversity and area on orthopteran species with contrasting mobility

Loss of semi-natural grasslands and reduction of habitat diversity are considered major potential threats to arthropod diversity in agricultural landscapes. The main aim of this study was to investigate how area and habitat diversity, mediated by shrub encroachment after grassland abandonment, affect species richness of orthopterans in island-like grasslands, and how contrasting mobility might alter species richness response to both factors. We selected 35 isolated patches in landscapes dominated by arable land (durum wheat) in order to obtain two statistically uncorrelated gradients: (i) one in habitat area ranging from 0.2 to 55 ha and (ii) one in habitat diversity ranging from patches dominated by one habitat (either open grasslands or shrublands) to patches with a mosaic of different habitats. Habitat loss due to land-use conversion into arable fields was associated with a substantial loss of species with a positive species-area relationship (SAR), with sedentary species having a steeper and stronger SAR than mobile species. Halting habitat loss is, therefore, needed to avoid further species extinctions. Shrub encroachment, triggered by abandonment, presented a hump-shaped relationship with habitat diversity. An increase in habitat diversity enhanced species richness irrespective of patch area and mobility. Maintaining or enhancing habitat diversity, by cutting or burning small sectors and by reintroducing extensive sheep grazing into abandoned grassland, are suggested as complementary strategies to mitigate further decline of orthopteran diversity in the remnant patches. This would be equally important in both small and large patches.     

Laboratory tests for diagnosing and monitoring canine leishmaniasis

Although several reviews on canine leishmaniasis have been published, none thoroughly described clinicopathologic abnormalities and their clinical usefulness. The aim of this review was to provide information concerning current diagnostic tests relevant for clinical pathologists and from a practical perspective. Specifically, in canine leishmaniasis, nonregenerative normocytic normochromic anemia, thrombocytopenia, or leukogram changes may be present. Clinical chemistry and urinalysis may indicate renal dysfunction (azotemia, decreased urine specific gravity, proteinuria) and an inflammatory/immune response (increased acute phase proteins [APP] or α₂‐ and/or γ‐globulins). Although a potential gammopathy is usually polyclonal, it may also appear oligo‐ or monoclonal, especially in dogs coinfected by other vector‐borne pathogens. When lesions are accessible to fine‐needle aspiration (lymphoadenomegaly, nodular lesions, joint swelling), cytology is strongly advised, as the presence of Leishmania amastigotes in a pattern of pyogranulomatous inflammation or lymphoplasmacytic hyperplasia is diagnostic. If the cytologic pattern is inconclusive, the parasite should be identified by histology/immunohistochemistry or PCR on surgical biopsies. Alternatively, cytology and PCR may be performed on bone marrow samples where amastigotes, along with erythroid hypoplasia, myeloid hyperplasia, plasmacytosis, or secondary dysmyelopoiesis can be observed. Dogs with overt leishmaniasis generally have high antibody titers, while low titers predominate in immunologically resistant infected dogs or in exposed dogs with no parasite confirmation. Quantitative serology is recommended in clinically suspect dogs as high‐titer antibodies titers may confirm the clinical diagnosis. In confirmed and treated dogs, renal function and inflammatory/immune response variables should be periodically monitored.     

Management strategy influences landscape patterns of high-severity burn patches in the southwestern United States

Landscape Ecology - Spatial patterns of high-severity wildfire in forests affect vegetation recovery pathways, watershed dynamics, and wildlife habitat across landscapes. Yet, less is known about...     

Comparative evaluation of live marker vaccine candidates "CP7_E2alf" and "flc11" along with C-strain "Riems" after oral vaccination

Due to the tremendous socio-economic impact of classical swine fever (CSF) outbreaks, emergency vaccination scenarios are continuously under discussion. Unfortunately, all currently available vaccines show restrictions either in terms of marker capacities or immunogenicity. Recent research efforts were therefore directed at the design of new modified live marker vaccines. Among the most promising candidates the chimeric pestiviruses "CP7_E2alf" and "flc11" were identified. Within an international research project, these candidates were comparatively tested in challenge experiments after a single oral vaccination. Challenge infection was carried out with highly virulent CSF virus strain "Koslov", 14 or 21 days post vaccination (dpv), respectively. Safety, efficacy, and marker potential were addressed. All assessments were done in comparison with the conventional "gold standard" C-strain "Riems" vaccine. In addition to the challenge trials, multiple vaccinations with both candidates were performed to further assess their marker vaccine potential. All vaccines were safe and yielded full protection upon challenge 21 days post vaccination. Neither serological nor virological investigations showed major differences among the three vaccines. Whereas CP7_E2alf also provided clinical protection upon challenge at 14 days post vaccination, only 50% of animals vaccinated with flc11, and 83% vaccinated with C-strain "Riems" survived challenge at this time point. No marked differences were seen in protected animals. Despite the fact that all multiple-vaccinated animals stayed sero-negative in the accompanying marker test, the discriminatory assay remains a weak point due to delayed or inexistent detection of some of the vaccinated and subsequently infected animals. Nevertheless, the potential as live marker vaccines could be confirmed for both vaccine candidates. Future efforts will therefore be directed at the licensing of "Cp7_E2alf" as the first live marker vaccine for CSF.     

Effects of glandular trichomes on the development of Phytophthora infestans infection in potato (S. tuberosum)

S. berthaulltii Hawkes, a wild potato species, possesses two types of glandular trichomes on its leaf surface: types A and B wich confer resistance to insects. Preliminary results indicated that the presence of glandular trichomes also confers resistance to Phytophthora infestans, the causal agent of late blight. In this paper we report the evaluation of trichome characteristics (density and exudate activities) performed on 8 lines of potato, including the control varieties Desirée and Tropicana, 5 hybrid lines derived from intercrosses and backcrosses between S. tuberosum and S. berthaultii and one accession of S. berthaultii. Whole plants of the 8 genotypes were inoculated with a spore suspension of Phytophthora infestans in order to evaluate their resistance to late blight. The resultant disease incidence correlated negatively with type A trichome density and with polyphenol-oxidase (PPO) activity of type A trichome glands. The removal of type B trichomes exudate by leaf dipping in ethanol did not influence the disease development after artificial inoculation of detached leaves. Type B trichome exudate was extracted and the fraction corresponding to Fatty Acids Sucrose Esters (FASE) was recovered after Thin Layer Chromatography and tested on the fungal mycelium in vitro. The FASE exhibited a slight inhibitory effect on mycelium growth. The results of the experiments are discussed, and several hypotheses regarding the possible role of types A and B trichomes are formulated. This revised version was published online in July 2006 with corrections to the Cover Date.     

A predator unmasked: life cycle of Bdellovibrio bacteriovorus from a genomic perspective

Predatory bacteria remain molecularly enigmatic, despite their presence in many microbial communities. Here we report the complete genome of Bdellovibrio bacteriovorus HD100, a predatory Gram-negative bacterium that invades and consumes other Gram-negative bacteria. Its surprisingly large genome shows no evidence of recent gene transfer from its prey. A plethora of paralogous gene families coding for enzymes, such as hydrolases and transporters, are used throughout the life cycle of B. bacteriovorus for prey entry, prey killing, and the uptake of complex molecules.     

Effect of soil drying on mucilage exudation and its water repellency: a new method to collect mucilage

Despite the importance of mucilage for soil–plant relations, little is known about the effect of soil drying on mucilage exudation. We introduce a method to collect mucilage from maize growing in wet and dry soils. Mucilage was collected from brace roots. The amount of mucilage exuded did not change with soil water content and transpiration rate. Mucilage exuded in dry soils had a higher degree of hydrophobicity, suggesting that the wetting properties of mucilage change in response to soil drying.