Development and evaluation of a highly sensitive and specific blocking enzyme-linked immunosorbent assay and polymerase chain reaction assay for diagnosis of bovine leukemia virus infection in cattle

OBJECTIVE: To develop a blocking ELISA for detection of bovine leukemia virus (BLV) antibodies that is comparable to a radioimmunoprecipitation (RIP) assay, to evaluate use of this ELISA for identification of BLV-infected herds, and to develop a polymerase chain reaction (PCR) assay for direct diagnosis of infection with BLV. SAMPLE POPULATION: Serum samples and pooled bulk-tank milk samples from cattle. PROCEDURE: The blocking ELISA was developed, using BLV gp51 as antigen, captured by a selected bovine polyclonal serum. A nested PCR was conducted with primers specific for a segment of the pol region of the BLV genome. RESULTS: Sensitivity and specificity of the ELISA were comparable to those of the RIP assay. Use of the ELISA on pooled milk samples allowed identification of herds in which prevalence of BLV infection among lactating cows was as low as 2.5%. Pooled milk samples from BLV-free herds did not react in the ELISA. All cattle that had positive results for the nested PCR had BLV antibodies, but cattle with consistantly low antibody titers required examination of sequential DNA samples to detect viral sequences. None of the 63 antibody-negative cattle had positive results for the PCR. CONCLUSIONS AND CLINICAL RELEVANCE: This ELISA is a highly specific and sensitive assay for the detection of BLV antibodies in serum and milk samples of cattle. Examination of pooled milk samples with the ELISA provides a reliable, practical, and economic procedure for identification of BLV-infected herds. The nested PCR also constitutes a specific procedure for direct diagnosis of infection with BLV.     

Antifungal activity of strawberry fruit volatile compounds against Colletotrichum acutatum

Eight volatile products characterizing strawberry aroma, which is generated from the oxidative degradation of linoleic and linolenic acids by a lipoxygenase (LOX) pathway, were examined because of their antifungal activity against Colletotrichum acutatum, one of the causal agents of strawberry anthracnose. In this study, the effects of aldehydes, alcohols, and esters on mycelial growth and conidia development were evaluated. (E)-Hex-2-enal was found to be the best inhibitor of mycelial growth [MID (minimum inhibitory doses)=33.65 microL L(-1)] and of spore germination (MID=6.76 microL L(-1)), while hexyl acetate was the least effective of all volatile compounds tested (MID=6441.89 microL L(-1) for mycelial growth and MID=1351.35 microL L(-1) for spore germination). Furthermore, the antifungal activity of (E)-hex-2-enal on susceptibility of strawberry fruits to C. acutatum was also confirmed. The presence of these molecules in jars containing strawberry fruits inoculated with a suspension of spores inhibited the fungus growth and prevented the appearance of symptoms. Moreover, a study of the effects of (E)-hex-2-enal on conidial cells of C. acutatum was evaluated by transmission electron microscopy. This volatile compound altered the structures of the cell wall and plasma membrane, causing disorganization and lysis of organelles and, eventually, cell death.     

Endoscopic removal of esophageal and ruminal foreign bodies in 5 Holstein calves

Endoscopic removal of esophageal and ruminal foreign bodies was successfully performed in 5 Holstein-Friesian calves under sedation or general anesthesia by using an electrocautery snare or a wire-guided Dormi basket. This report describes the endoscopic manipulations, treatment, and outcomes of esophageal foreign body removal in these calves.     

Using fishers' anecdotes, naturalists' observations and grey literature to reassess marine species at risk: the case of the Gulf grouper in the Gulf of California, Mexico

Designing fishing policies without knowledge of past levels of target species abundance is a dangerous omission for fisheries management. However, as fisheries monitoring started long after exploitation of many species began, this is a difficult issue to address. Here we show how the ‘shifting baseline’ syndrome can affect the stock assessment of a vulnerable species by masking real population trends and thereby put marine animals at serious risk. Current fishery data suggest that landings of the large Gulf grouper (Mycteroperca jordani, Serranidae) are increasing in the Gulf of California. However, reviews of historical evidence, naturalists’ observations and a systematic documentation of fishers’ perceptions of trends in the abundance of this species indicate that it has dramatically declined. The heyday for the Gulf grouper fishery occurred prior to the 1970s, after which abundance dropped rapidly, probably falling to a few percent of former numbers. This decline happened long before fishery statistics were formally developed. We use the case of the Gulf grouper to illustrate how other vulnerable tropical and semi‐tropical fish and shellfish species around the world may be facing the same fate as the Gulf grouper. In accordance with other recent studies, we recommend using historical tools as part of a broad data‐gathering approach to assess the conservation status of marine species that are vulnerable to over‐exploitation.     

Drivers for spatial modelling of a critically endangered seabird on a dynamic ocean area: Balearic shearwaters are non-vegetarian

1. Spatial modelling is an important research tool to improve our knowledge about the distribution of wildlife in the ocean. Using different modelling techniques (MaxEnt and a generalized linear mixed model), a predictive habitat suitability model was developed for one of the most threatened seabirds in the world: the Balearic shearwater, Puffinus mauretanicus.
2. Models were developed using a 10-year dataset from the Gulf of Cádiz (on the south-western Iberian Peninsula), a key foraging area for Balearic shearwaters during migration and the non-breeding season.
3. Predictive habitat maps strongly matched the observed distribution patterns, pointing to bathymetric features as the main modelling drivers. The species was concentrated on shallow areas (up to approximately 100 m in depth) of the continental shelf, very close to the mouth of the Guadalquivir River. In contrast with previous studies, Balearic shearwater distribution in the highly dynamic Gulf of Cádiz was not correlated with areas of high chlorophyll a concentration.
4. This lack of spatial correlation probably arises from the delay between the phytoplankton bloom and the response of the zooplankton and small fish that are preyed upon by Balearic shearwaters, which may result in important displacements of this trophic chain across the Gulf of Cádiz.
5. The analysis presented contributes to a better understanding of the spatial distribution and ecology of the critically endangered top predator in the Gulf of Cádiz and offers important information to improve management plans.

     

Differences in the diversity of oribatid mite communities in forests and agrosystems lands

A field study was conducted in the province of Burgos (Castile, NW of Spain) to assess the role and influence of the different anthropic use of soil on the oribatid mite communities (Acari, Oribatida).Soil samples from 20 representative soil sites of the Castilian upland steppes, with cultured, pastureland or forest soil uses, were taken in spring and autumn 2000. Thereafter, soil samples were analysed in terms of their mesofaunal biodiversity, measured using the real and relative diversity of the oribatid mite communities.Collected individuals were identified to species level, being altogether 111 Oribatid species. Results from communities' ordination analysis showed clear gradients based on community variables. One of these was defined by plots with high values of diversity and species richness which correspond to the less anthropic natural soils, mainly oak forest. In contrast, the most anthropic agrosystems soils, including extensive cereal crops, most of them receiving only mineral fertilisation or having human management, were placed in opposite gradients. Differences in biodiversity between crop lands and natural soils were also confirmed by ANOVA. The seasonality, measured in terms of difference between spring and autumn plots, played a minimum role in explaining differences of diversity. Nevertheless autumn diversity values were slightly higher than those of spring, except for crops.     

Reaction of B. cereus bacteria and peroxidase enzymes under pressures >400 MPa.

It is known that B. cereus (Gram-positive bacteria) and peroxidase enzymes are resistant to pressures of approximately 400 MPa in fruit and vegetable products among others. The aim of the present work is to have knowledge about their behavior when using pressures >400 MPa without other combined treatments. The results showed that B. cereus was not inactivated at pressures of 1000 MPa for 15 min at 20 degrees C. In peroxidase enzymes the results remained similar and the pressure of 1000 MPa for 30 min was not enough to inactivate them. The apple cell structure at these high-pressure levels revealed that it changed and the cells were less cemented. The treated apple presented a translucent aspect, and some fluids migrated from the inside to the outside of the cell.