Contrasting effects of heat shock during in vitro maturation on development of in vitro‐fertilized and parthenogenetic bovine embryos

This study investigated the influence of heat shock during in vitro maturation on embryo development following in vitro fertilization (IVF) or parthenogenesis (Part). Immature bovine cumulus–oocyte complexes were exposed to heat shock (41.0°C) during the first 12 hr of in vitro maturation (IVM), followed by 12 hr at 38.5°C. Control group consisted of in vitro maturation for 24 hr at 38.5°C. Oocytes were in vitro‐fertilized or activated with ionomycin and cultured in vitro for 192 hr post‐in vitro insemination or parthenogenetic activation (hpia). There was an interaction (p < .01) between temperature of IVM and method of oocyte activation (IVF or Part) for cleavage at 48 hpia. Heat shock had a negative impact (p < .01) on cleavage of IVF embryos, whereas no (p > .05) effect was found in the Part embryos. Embryo development towards blastocyst stage at 168 and 192 hpia decreased in both IVF and Part embryos derived from heat‐shocked oocytes. Heat shock increased (p < .05) the apoptotic index in Part blastocysts, but no effect (p > .05) was found in IVF counterparts. Heat shock also down‐regulated the expression of AQP3 (p < .01) and up‐regulated the expression of HSP70.1 (p < .01) in Part blastocysts, whereas it down‐regulated the expression of ATP1A1 (p < .05) in IVF blastocysts. In conclusion, the effects of heat shock during IVM on early embryo cleavage and blastocyst apoptosis are influenced by the method of oocyte activation and expression of some genes can be disturbed in embryos derived from heat‐shocked oocytes.     

Conjunctival fungal flora in healthy donkeys

OBJECTIVE: To identify and quantify ocular fungi from healthy donkeys living in the center of Italy. ANIMALS STUDIED: One hundred and two Amiata donkeys were examined. PROCEDURES: Conjunctival swabs from both eyes were seeded onto Sabouraud dextrose agar (SDA) and malt extract agar (MEA), and incubated at 25 degrees C over a 10-day period. Filamentous fungi identification was achieved to the genus level; yeast colonies were identified for macro-micromorphologic and physiological characteristics. RESULTS: Eighty-one donkeys out of 102 (79.4%) were positive for fungi; 47/102 (46.1%) had positive cultures from both eyes. Most frequently recovered fungal genera were Aspergillus spp., Penicillium spp., Cladosporium spp., Acremonium spp. Different fungal genera and/or species were recovered from the same donkey in 43 cases (42.1%). Yeasts were isolated from five subjects; the yeasts were never associated with molds. The number of colony forming units (CFU) ranged from 1 to 100. CONCLUSIONS: Aspergillus was the most commonly isolated fungal genus (33%). This result agrees with the findings of similar surveys carried out in horses. There was a remarkable presence of fungi and perfect forms. These observations may be explained by the optimal conditions for presence and development of fungi in the conjunctival fornix microenvironment in Amiata donkeys.     

Progress and challenges in freshwater conservation planning

• 1. Freshwater ecosystems and their associated biota are among the most endangered in the world. This, combined with escalating human pressure on water resources, demands that urgent measures be taken to conserve freshwater ecosystems and the services they provide. Systematic conservation planning provides a strategic and scientifically defensible framework for doing this.
• 2. Pioneered in the terrestrial realm, there has been some scepticism associated with the applicability of systematic approaches to freshwater conservation planning. Recent studies, however, indicate that it is possible to apply overarching systematic conservation planning goals to the freshwater realm although the specific methods for achieving these will differ, particularly in relation to the strong connectivity inherent to most freshwater systems.
• 3. Progress has been made in establishing surrogates that depict freshwater biodiversity and ecological integrity, developing complementarity‐based algorithms that incorporate directional connectivity, and designing freshwater conservation area networks that take cognizance of both connectivity and implementation practicalities.
• 4. Key research priorities include increased impetus on planning for non‐riverine freshwater systems; evaluating the effectiveness of freshwater biodiversity surrogates; establishing scientifically defensible conservation targets; developing complementarity‐based algorithms that simultaneously consider connectivity issues for both lentic and lotic water bodies; developing integrated conservation plans across freshwater, terrestrial and marine realms; incorporating uncertainty and dynamic threats into freshwater conservation planning; collection and collation of scale‐appropriate primary data; and building an evidence‐base to support improved implementation of freshwater conservation plans.

Copyright © 2008 John Wiley & Sons, Ltd.     

Molecular phylogenetic,morphological, and pathogenic analyses reveal a single clonal population of Septoria lycopersici with a narrower host range in Brazil

Septoria leaf spot, caused by Septoria lycopersici, is considered one of the most important diseases of tomato in Brazil. Despite its importance, the disease agent is still poorly studied. Septoria isolates collected from different production regions of Brazil were characterized by molecular, morphological, and pathogenic methods. A set of 104 isolates was sequenced for the DNA Tub, Cal, and EF1-α loci. Ten isolates were selected, according to geographical region of origin and type of leaf lesion (typical or atypical), for morphological characterization and for evaluation of aggressiveness on tomato cultivar Santa Clara. To evaluate the pathogen host range, cultivated and wild Solanaceae plants were inoculated with four selected isolates. The results showed that all isolates grouped with the type isolate of S. lycopersici in maximum likelihood and Bayesian inference trees. The isolates were morphologically similar. All isolates selected for pathogenicity testing on tomato were able to induce typical symptoms of the disease, but differed in their aggressiveness. A total of eight species of Solanaceae were also identified as potential alternative hosts for S. lycopersici. This information will provide a more accurate assessment of the risks involved with the introduction of new crops, especially of the genus Solanum, in areas where the species is already present. In addition, it will provide the basis for the establishment of more efficient methods in the management of Septoria leaf spot of tomatoes in natural conditions and in the different production systems.     

Interception of water by pastures of Pennisetum clandestinum Hochst. ex Chiov. and Melinis minutiflora Beauv.

The conversion of montane cloud forests into pastures for grazing cattle is the main cause of important impacts on rivers and streams in most of the Andes. In order to evaluate the effects of vegetation changes we need to understand water fluxes, particularly canopy interception. We measured net precipitation responses by the canopies of Pennisetum clandestinum Hochst. ex Chiov. and Melinis minutiflora Beauv. under simulated rainfall. The samples were taken from grazed pastures during 1 year, digging out round mats of grass, placing them on wire-mesh devices after eliminating soil and underground biomass, and irrigating with different water amounts. The resulting data allow us to present a validated model for each species that predicts percentage of interception at different precipitation intensities taking into account previous canopy wetness within determined biomass ranges. We use these models to estimate 2 years of interception values for pastures of both species in the upper watershed of El Cañadón, Capaz River, Venezuelan Andes. Mean annual precipitation of El Cañadón is 1244 mm and the estimate of pasture interception was 36.5% and 31.8% for P. clandestinum and M. minutiflora, respectively. Interception models, such as the ones derived in this study, provide a basis for quantifying interception rates as a function of previous wetness of canopies and grass species.     

Soil chemical and biochemical properties of a salt-marsh alluvial Spanish area after long-term reclamation

Marisma, one of the largest salt-marsh alluvial areas in SW Spain, has been reclaimed since 1970 by artificial drainage and amendment with phosphogypsum (PG) so as to reduce Na⁺ saturation. Within the reclaimed area, two 250- × 20-m plots were treated as follows: (1) amendment with 25 Mg/ha of PG every 2 to 3 years between 1979 and 2003 (plot PY); (2) like PY but PG treatment stopped after 1997 (plot DR). A contiguous virgin Marisma salt-marsh plot (MV), neither drained nor amended, was the control. In MV, soil microbial biomass C, most enzyme activities and total organic C content were much greater than in PY and DR soils, despite the salinity stress. The decrease in soil organic matter content in PY and DR soils was likely due to cotton-cropping practices, which favoured the organic C mineralisation and nutrients removal by crops. Microbial activity of MV soil, probably stimulated by the rhizodepositions of the natural vegetation, did not suffer from the osmotic effect due to the raising of soil solution ionic strength. Microbial quotient could be ranked as MV > PY > DR, whereas the metabolic quotient had an opposite trend. Thus, both quotients suggested that the interruption of PG amendment was not favouring microbial activity. Principal component analysis clearly identified the chemical and biochemical soil properties mostly affected by the reclamation and also indicated the longer PG amendment in PY plot. Stepwise discriminant analysis identified two physiologically different types of soil microflora, one less active present in the MV virgin soil and another more active present in the reclaimed PY and DR soils.     

Analysis of weight uniformity, content uniformity and 30-day stability in halves and quarters of routinely prescribed cardiovascular medications

Objectives

Congenital and acquired cardiac disorders are frequently diagnosed in small breed dogs and cats. In order to appropriately dose cardiovascular drugs for small patients, fractions of commercially available tablets must be prescribed. The aims of this study were to evaluate weight and content uniformity and 30-day chemical stability in halves and quarters of 11 drug formulations commonly prescribed to treat cardiovascular disorders in small breed dogs and cats.

Animals, materials and methods

Fifteen tablets from 11 drug formulations were obtained within the same lot. Tablets were split by a single operator using a commercially available pill splitter. Whole tablets, halves and quarters were weighed and stored in plastic containers. High-pressure liquid chromatography or liquid chromatography with mass spectrometry were utilized to determine drug content and repeated 30 days later to estimate chemical stability.

Results

Statistically significant weight variability, content non-uniformity and chemical degradation were found for some formulations. Digoxin showed a significant degradation that should be considered in clinical practice.

Conclusions

It appears that pill splitting is overall a relatively reliable practice; however tablets should not be split ahead of time but only immediately prior to intended usage.