Polymorphism of 14α-demethylase Gene (CYP51) in the Cereal Eyespot Fungi Tapesia acuformis and Tapesia yallundae

Cereal eyespot fungi Tapesia acuformis and Tapesia yallundae are closely related species which show different behaviours upon treatment with sterol 14-demethylase inhibitors (DMIs). T. acuformis is naturally resistant to DMIs belonging to the triazole family and susceptible to the imidazole ones, whilst T. yallundae is sensitive to both inhibitors. Cloning of the target enzyme gene, CYP51, from the two species revealed an important polymorphism between them. Further sequencing of CYP51 from sixteen T. acuformis and eleven T. yallundae strains with different phenotypes with regards to resistance to DMIs confirmed that at least eleven variations are species related. Among them, a conserved phenylalanine residue at position 180, found both in T. yallundae and in all known CYP51 proteins from filamentous fungi and yeast, was replaced in T. acuformis by a leucine. Therefore, a leucine at 180 could be possibly involved in natural resistance of T. acuformis to triazoles. Other mutations were observed in some resistant strains, sometimes simultaneously, but in contrast to what was reported for other filamentous fungi, where a mutation at the 136 position of the CYP51 gene product seemed to correlate with resistance to DMIs, we did not find a clear relationship between a given mutation and a particular phenotype. This result suggests that resistance to DMIs could have a polygenic nature in Tapesia. We took advantage of species-related variations to develop a PCR-based assay allowing rapid and easy discrimination between field strains of the two species.     

Metabolism of benomyl in carrot,strawberry and apple

The metabolism of benomyl was studied in carrots treated with high rates of benomyl, and in strawberries and apples treated at normal rates. The same metabolites were observed in both cases though, at normal rates, their concentrations were low but significant. Detected by means of gas-liquid chromatography they were: carbendazim, 2-aminobenzimidazole (2-AB), benzimidazole, 2-aminobenzonitrile, o-phenylene-diamine and the conjugates of carbendazim and 2-AB. o-Phenylenediamine has not hitherto been reported as a metabolite of benomyl. Two unidentified compounds were also observed by gas-liquid chromatography; their mass spectra were obtained by means of combined gas-liquid chromatography and mass spectrometry.     

Comparison of the pig and feline models for full thickness corneal transplantation

Purpose The goal of this study was to report on the advantages and limitations of the pig and feline models for experimental in vivo corneal transplantation. Methods Ten healthy domestic pigs and ten healthy cats were used. Full thickness penetrating keratoplasty was performed using autologous (eight cases), allogeneic (seven cases) or human xenogeneic (three cases) tissue. In two other cases, the inflammatory response to partial thickness trephination (without transplantation) was evaluated. Eyes were assessed daily before and after surgery by slit‐lamp, pachymetry, and tonometry. A transparency score ranging from 0 (opaque graft) to 4 (clear graft) was used, based on the slit‐lamp examination. Optical coherence tomography, histology, and electron microscopy were performed postmortem. Results In the pig, the mean (±SD) transparency score for the eight full thickness grafts was 0.88 ± 0.99, ranging from 0 to 3. In the feline model, the mean transparency score for the seven uncomplicated grafts was 3.93 ± 0.19, ranging from 3.5 to 4. Both negative controls without endothelium remained opaque at all time. Intraoperative tendency for iris incarceration into the wound, rapid corneal swelling, suture cheese wiring, and postoperative intraocular inflammation were the main factors jeopardizing the functional success of the corneal transplant in the pig model. Conclusion Suboptimal functional results were obtained after full thickness corneal transplantation in the pig model, while in the feline model, the same protocol yielded uneventful surgeries and clear transplants, with functional results similar to those achieved in human subjects.     

Immunohistochemical Expression of Growth Factors in the Follicular Wall of Normal and Cystic Ovaries of Sows

The expression of growth factors was evaluated immunohistochemically in normal and cystic ovaries of sows. The immunohistochemically stained area (IHCSA) was quantified by image analysis to analyse the expression of these proteins in the follicular wall of secondary, tertiary and cystic follicles. IGF‐I immunoreactivity was strong in the granulosa cell layer (GC), moderate in the theca interna (TI) and mild in the theca externa (TE) of the normal follicles. There was severe reduction of the labelling to IGF‐I in the GC of the follicular and luteinized cysts. In the normal follicles, the reactivity for IGF‐II was very similar to pattern noted in IGF‐I. There was reduction of the IHCSAs in the GC of the follicular and luteinized cysts, but the decrease was not significant. The staining of the IGF‐II in the TI and TE of the cysts was increased, in comparison with normal follicles. The IHCSAs for VEGF were higher in the GC and TE of the normal follicles in contrast to TI, but this difference was noted only in the tertiary follicle. The VEGF reactivity increased in the GC of the cysts, in relation to normal follicles. The results of the current study show that the formation of ovarian cysts in sows is associated with alterations in the immunohistochemical expression of some growth factors.     

Impact of land degradation on soil respiration in a steppe (Stipa tenacissima L.) semi-arid ecosystem in the SE of Spain

Climate change scenarios predict increases in temperature, changes in precipitation patterns, and longer drought periods in most semi-arid regions of the world. Ecosystems in these regions are prone to land degradation, which may be aggravated by climate change. Soil respiration is one of the main processes responsible for organic carbon losses from arid and semi-arid ecosystems. We measured soil respiration over one year in two steppe ecosystems having different degrees of land degradation under three ground-covers: with vegetation, bare soil, and an intermediate situation between plants and bare soil.The largest differences in soil respiration rates between the sites were observed in spring, coinciding with the highest level of plant activity. The degraded site had drier and hotter soils with less soil water availability and a longer drought period. As a result, vegetation on the degraded site did not respond to spring rainfall events. Soil respiration showed a strong seasonal variability, with average annual rates of 1.1 and 0.8 μmol CO₂ m⁻² s⁻¹ in the natural and degraded sites, respectively. We did not observe significant differences in soil respiration rates associated with ground-cover i.e., the temporal variation was much larger than the spatial variation. At both sites, soil moisture was the controlling driver of soil respiration for most of the year, when temperatures were above 20 °C and constrained the response to temperature for the few months when the temperature was below 20 °C. An empirical model based on soil temperature and soil moisture explained 90% and 72% of the seasonal variability of soil respiration on the natural and degraded sites, respectively. For the first time, this study suggests that land degradation may alter the carbon balance of these ecosystems through changes in the temporal dynamics of soil respiration and plant productivity, which have important negative consequences for ecosystem functioning and sustainability.     

Carbon mineralization rates at different soil depths across a network of European forest sites (FORCAST)

Most of the carbon (C) in terrestrial ecosystems is stored in the mineral soil layers. Thus, the response of the mineral soil to potential increases in temperature is crucial for the prediction of the impact of climate change on terrestrial ecosystems. Samples from three mineral soil layers were collected from eight mature forest sites in the European network CARBOEUROFLUX and were incubated at four temperatures (4, 10, 20 and 30°C) for c. 270 days. Carbon mineralization rates were related to soil and site characteristics. Soil water holding capacity, C content, nitrogen (N) content and organic matter all decreased with soil depth at all sites, with significantly larger amounts of organic matter, C and N in the top 0–5 cm of mineral soil than in the deeper layers. The conifer forest soils had significantly lower pH, higher C/N ratios and carbon contents in the top 5 cm than the broadleaf forest soils. Carbon mineralization rates decreased with soil depth and time at all sites but increased with temperature, with the highest rates measured at 30°C for all sites. Between 50 and 70% of the total C respired after 270 days of incubation came from the top 5 cm. The percentage C loss was small in all cases, ranging from 1 to 10%. A two‐compartment model was fitted to all data to derive the labile/active and slow/recalcitrant fractions, as well as their decomposition constants. Although the labile fraction was small in all cases, we found significantly larger amounts of labile C in the broadleaf forest soils than in the conifer forest soils. No statistically significant differences were found in the temperature sensitivity parameter Q₁₀ among sites, soil layers or between conifer and broadleaf soils. The average Q₁₀ for all soils was 2.98 (± 0.10). We found that despite large differences among sites, C mineralization can be successfully predicted as a combined function of site leaf area index, mean annual temperature and content of labile carbon in the soil (R² = 0.93).     

Soil properties and the macrofauna community in abandoned irrigated rice fields of northeastern Argentina

This study compared soil physical, chemical, and biological characteristics between natural grassland and recently abandoned rice fields in order to identify those variables that might explain the observed increase of Camponotus punctulatus anthills in abandoned rice paddy fields from Northern Argentina. Mainly due to a reduction of macropores and mesopores, overall porosity decreased by around 6% and bulk density was about 7% greater, in the 0- to 10- and 10- to 20-cm layers of the abandoned rice fields. Carbon and nitrogen content from organic matter increased (29% and 41% respectively for the 0- to 20-cm horizon) during cultivation but decreased (38% and 24%) 2 years after the last rice harvest. Forty percent of natural grassland-organic matter and 30% of abandoned rice-organic matter mineralized in less than 2 years. There was a different community structure between the abandoned rice fields and the undisturbed natural grassland and only a 20.6% (i.e. only 19 species from a total of 92) overlap in species composition. The abundance of macrofauna was greater in abandoned rice fields (2,208 individuals m^–2) in comparison to natural grasslands (288 ind m^–2) due to higher densities of small earthworms and Camponotus punctulatus ants; however, the Shannon index showed lower values in comparison to natural grasslands. Earthworms and C. punctulatus in the abandoned rice fields showed a change in their ¹³C signature indicating a switch in diet from natural grassland organic matter (C4) to organic matter from rice (C3). Our results indicate that the effects of rice cultivation practices did not seem to produce any physical or trophic limitations to recolonization by the macrofauna. It seems that changes in overall soil conditions have favored a change in the construction behavior of C. punctulatus which, in combination with population increases, could explain the explosion in number of anthills.     

Soil microbial community characteristics along an elevation gradient in the Laguna Mountains of Southern California

We sampled soil at four sites in the Laguna Mountains in the western Sonoran Desert to test the effects of site and sample location (between or beneath plants) on fatty acid methyl ester (FAME) and carbon substrate ulilization (Biolog) profiles. The four sites differed in elevation, soil type, plant community composition, and plant percent cover. Soil pH decreased and plant density increased with elevation. Fertile islands, defined as areas beneath plants with greater soil resources than bare areas, are present at all sites, but are most pronounced at lower elevations. Consistent with this pattern, fertile islands had the greatest influence on FAME and Biolog profiles at lower elevations. Based on the use of FAME biomarker and principal components analyses, we found that soil microbial communities between plants at the lowest elevation had proportionally more Gram-negative bacteria than all other soils. At the higher elevation sites there were few differences in FAME profiles of soils sampled between vs. beneath plants. Differences in FAME profiles under plants among the four sites were small, suggesting that the plant influence per se is more important than plant type in controlling FAME profiles. Since microbial biomass carbon was correlated with FAME number (r=0.85,P<0.0001) and with FAME named (r=0.88,P<0.0001) and total areas (r=0.84,P<0.0001), we standardized the FAME data to ensure that differences in FAME profiles among samples were not the result of differences in microbial biomass. Differences in microbial substrate utilization profiles among sampling locations were greatest between samples taken under vs. between plants at the two lower elevation sites. Microbial substrate utilization profiles, therefore, also seem to be influenced more by the presence of plants than by specific plant type.     

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

Small ruminant lentiviruses (SRLV = maedi-visna in sheep and caprine arthritis encephalitis in goats) are distributed throughout most countries of the world, particularly Europe. Laboratories from 16 European countries established collaborations within the framework of a COST (CO-operation in the field of Scientific and Technical Research) action sponsored by the European Union in order to (i) better organize their research programmes on SRLVs and (ii) to coordinate efforts to combat these two diseases. After five years, a consensus conference--the first one in the veterinary medicine field--concluded the work of this network of laboratories by reviewing the present position and discussing three important questions in the field of SRLVs: routes of transmission, consequences of infection and potential role of eradication programmes at either a European or local level, according to the situation in each country or region. This paper brings together existing information regarding these questions and identifies areas for future research.