Diversity of laccase genes from basidiomycetes in a forest soil

Fungal oxidative exo-enzymes lacking substrate specificity play a central role in the cycling of soil organic matter. Due to their broad ecological impact and available knowledge of their gene structure, laccases appeared to be appropriate markers to monitor fungi with this kind of oxidative potential in soils. A degenerate PCR-primer pair Cu1F/Cu2R, specific for basidiomycetes, was designed to assess directly the diversity of laccase genes in soils. PCR amplification of mycelial cultures and fruit-bodies of a wide spectrum of basidiomycetes, covering all functional groups (saprophytes, symbionts, and pathogens), produced multiple DNA fragments around 200 bp. A neighbor-joining tree analysis of the PCR-amplified laccase sequences showed a clear species-specificity, but also revealed that most fungal taxa possess several laccase genes showing a large sequence divergence. This sequence diversity precluded the systematic attribution of amplified laccase of unknown origin to specific taxa. Amplification of laccase sequences from DNA, extracted from a brown (moder) forest soil, showed a specific distribution of laccase genes and of the corresponding fungal species in the various soil horizons (O_h, A_h, B_v). The most organic O_h-horizon displayed the highest gene diversity. Saprophytic fungi appeared to be less widespread through the soil horizons and displayed a higher diversity of laccase genes than the mycorrhizal ones.     

Methionine oxidation enhances κ-casein amyloid fibril formation

The effects of protein oxidation, for example of methionine residues, are linked to many diseases, including those of protein misfolding, such as Alzheimer's disease. Protein misfolding diseases are characterized by the accumulation of insoluble proteinaceous aggregates comprised mainly of amyloid fibrils. Amyloid-containing bodies known as corpora amylacea (CA) are also found in mammary secretory tissue, where their presence slows milk flow. The major milk protein κ-casein readily forms amyloid fibrils under physiological conditions. Milk exists in an extracellular oxidizing environment. Accordingly, the two methionine residues in κ-casein (Met(95) and Met(106)) were selectively oxidized and the effects on the fibril-forming propensity, cellular toxicity, chaperone ability, and structure of κ-casein were determined. Oxidation resulted in an increase in the rate of fibril formation and a greater level of cellular toxicity. β-Casein, which inhibits κ-casein fibril formation in vitro, was less effective at suppressing fibril formation of oxidized κ-casein. The ability of κ-casein to prevent the amorphous aggregation of target proteins was slightly enhanced upon methionine oxidation, which may arise from the protein's greater exposed surface hydrophobicity. No significant changes to κ-casein's intrinsically disordered structure occurred upon oxidation. The enhanced rate of fibril formation of oxidized κ-casein, coupled with the reduced chaperone ability of β-casein to prevent this aggregation, may affect casein-casein interaction within the casein micelle and thereby promote κ-casein aggregation and contribute to the formation of CA.     

Further Contributions to the Development of a Differential Set of Pea Cultivars (Pisum sativum) to Investigate the Virulence of Isolates of Aphanomyces euteiches

Common root rot (Aphanomyces euteiches Drechs.) has become a very destructive disease in the French pea crops since 1993. For an accurate investigation of the virulence variability among French A. euteiches populations and between French and foreign populations, a new set of differential pea genotypes was developed. Thirty-three American and European pea lines, displaying different levels of resistance, were screened in a growth chamber against two French isolates. Symptoms (disease severity from 0 to 5, evaluating symptom surface on roots and epicotyl) and percentage of top fresh weight (inoculated/uninoculated top fresh weight ratio) were measured. From this screening 12 relatively resistant lines, from various genetic backgrounds, were identified along with a highly susceptible control. This set of 13 genotypes was inoculated under controlled conditions with 14 isolates from France, Sweden, USA, Canada and New Zealand, to investigate genotype–isolate interactions. Root symptoms were rated (disease severity), and a susceptibility/resistance threshold was established at disease severity = 1. Significant quantitative interactions were observed, and five 'resistance patterns' were identified, leading to a set of six pea genotypes: Baccara (susceptible), Capella, MN313, 902131, 552 and PI180693. Fields trials of this set in 1999 and 2000 gave the same resistance rankings than in growth chamber conditions. This set will allow more accurate assessments of the variability in virulence/aggressiveness of A. euteiches isolates from France and foreign countries, and further investigations of the epidemiological and genetic basis of pea–A. euteiches interactions.     

Target and non-target site mechanisms of fungicide resistance and their implications for the management of crop pathogens

Fungicides are indispensable for high-quality crops, but the rapid emergence and evolution of fungicide resistance have become the most important issues in modern agriculture. Hence, the sustainability and profitability of agricultural production have been challenged due to the limited number of fungicide chemical classes. Resistance to site-specific fungicides has principally been linked to target and non-target site mechanisms. These mechanisms change the structure or expression level, affecting fungicide efficacy and resulting in different and varying resistance levels. This review provides background information about fungicide resistance mechanisms and their implications for developing anti-resistance strategies in plant pathogens. Here, our purpose was to review changes at the target and non-target sites of quinone outside inhibitor (QoI) fungicides, methyl-benzimidazole carbamate (MBC) fungicides, demethylation inhibitor (DMI) fungicides, and succinate dehydrogenase inhibitor (SDHI) fungicides and to evaluate if they may also be associated with a fitness cost on crop pathogen populations. The current knowledge suggests that understanding fungicide resistance mechanisms can facilitate resistance monitoring and assist in developing anti-resistance strategies and new fungicide molecules to help solve this issue. © 2023 Society of Chemical Industry.     

Exploiting a readily available but hard to digest resource: A review of exudativorous mammals identified thus far and how they cope in captivity

Gum is a widely available carbohydrate, composed mainly of non‐digestible structural carbohydrates. No mammalian enzymes can digest gum; therefore, a mammal ingesting gum must rely on microbial fermentation to access the energy it possesses. Gums are relatively nutrient poor. Despite this, some mammals have evolved to exploit this food resource. We aim to review the literature for all mammal species which have been recorded to ingest gum, whether quantified or not, and discuss this in the context of their evolutionary adaptations. We also investigated the recommended captive diets for these species to look at whether gum is recommended. We conducted a literature search on ISI Web of Knowledge to tabulate all mammal species observed ingesting gum and classified them as obligate, facultative or opportunistic feeders. We encountered 94 mammal species that eat gum in the wild (27 obligate feeders, 34 facultative feeders and 33 opportunistic feeders). Obligate feeders have entirely evolved to exploit this resource but were found to not be given gum in captivity, which may explain why they are failing to thrive, as opposed to facultative feeders, which have fewer issues. Gum may be necessary for the health of obligate feeders in captivity. Future research should focus on the physiological effects that gum ingestion poses on different digestive systems.     

Formation of volatile organic products in soils under anaerobiosis—I: Metabolism of glucose

A simple trapping procedure for concentration of volatile organic compounds present in a head space and the subsequent analysis of the retained compounds by combined gas chromatographymass spectrometry is described. The procedure was used to study the organic metabolites evolved from various soils when they were amended with glucose and incubated under argon. A number of organic acids, alcohols, aldehydes and esters were evolved during a 4 week period. Less than 10 per cent of the added carbon was recovered as organic compounds, which was largely due to poor recovery of volatile organic acids from soil. Some implications of the observations to microbial ecology are discussed.     

Early dexamethasone treatment after implantation of a sciatic-nerve cuff decreases the concentration of substance P in the lumbar spinal cord of rats with neuropathic pain

The main purpose of this study was to evaluate the effects of early dexamethasone treatment on pain-related peptides at an early stage in the development of neuropathic pain induced by implantation of a sciatic nerve cuff in Sprague Dawley rats (body weight 250 to 350 g). The rats were tested for touch sensitivity with the use of von Frey filaments before and 3 d after cuff implantation (n = 12) or sham surgery (n = 6). Half of the cuff-implanted rats received dexamethasone, 1 mg/kg intraperitoneally, 1 h after surgery. Spinal cords were collected on the 3rd day after surgery, and the lumbar enlargement was processed for the detection of selected peptides (neurotensin, substance P, cholecystokinin [CCK], vasoactive intestinal peptide, and calcitonin gene-related peptide) by means of liquid chromatography and tandem mass spectrometry. The right sciatic nerve of each rat was collected, fixed, and stained for histopathological evaluation. Except for neurotensin, all the peptides showed an increased concentration with neuropathic pain; however, the differences were significant (P < 0.05) only for substance P and CCK. In the animals treated with dexamethasone, mechanical allodynia was less pronounced (P < 0.01), and only the concentration of substance P was decreased significantly (P < 0.05). Sciatic nerve sections showed a decrease in C (P < 0.01) and Adelta (P < 0.03) fibres with neuropathic pain and a nearly normal percentage of C fibres after dexamethasone treatment. The dexamethasone-treated animals also had less inflammation detectable microscopically at the nerve constriction site compared with cuff-implanted animals that were not treated with dexamethasone. Our results suggest that in the early stages of neuropathic pain induced by an inflammatory process, dexamethasone may be a useful treatment and that substance P plays an important role in pain perception.     

Risk factors associated with prevalence of tuberculosis-like lesions and associated mycobacteria in cattle slaughtered at public and export abattoirs in Ethiopia

This study was aimed at investigating risk factors associated with prevalence of tuberculosis (TB)-like lesions and associated mycobacteria in Ethiopian cattle slaughtered. The study was carried out during 2006–2007 in five selected municipal and export abattoirs. Methods of investigation involved detailed necropsy examination of carcasses and isolation of mycobacteria from pathologic tissue specimens. Factors of animal and environment origin were recorded and examined as explanatory variables in relation to the presence or absence of TB-like lesions and the presence of viable mycobacteria. Multivariable logistic regression analysis was used to identify risk factors associated with prevalence of TB-like lesions and mycobacteria. Out of 3,322 carcasses inspected, 10.2% (95% confidence interval (CI) [9.2–11.3]) and 3.2% (95% CI [2.6–3.8]) were positive, respectively, based on necropsy and bacteriologic examinations. The highest and lowest lesion prevalence was recorded in Adama (24.7%) and Yabello (4.2%), respectively. Multivariable logistic regression analysis identified age, breed, abattoir location, geographic origin and management system as being risk factors for prevalence of TB-like lesions and occurrence of viable mycobacteria in Ethiopian cattle. The study demonstrated mycobacterial infection as important infectious disease of Ethiopian cattle. The reported confirmed cases of the disease in different management and geographic settings in Ethiopia disproved the earlier held opinion of its occurrence as a low sporadic profile. In view of a dietary proclivity of Ethiopian communities (milk and meat are predominantly consumed raw) and lifestyle (close contact of people with animals), the risk of bovine tuberculosis as a public health threat is eminent.     

Serum alpha globulin fraction: survival-and-recovery effect in irradiated mice

An alpha macroglobulin fraction (19S) was isolated from the serum of rats and BC(3)F(1) mice by zonal ultracentrifugation. Both the isologous and heterologous macroglobulin fractions increased survival among BC(3)F(1) mice x-irradiated with 750 roentgens. The mouse macroglobulin fraction also enhanced radiation recovery of hematopoietic tissue as measured by colony-forming assay and iron-59 incorporation into erythropoietic cells. The overall difference in hematopoietic activity in the irradiated (400 roentgens) mice treated with the macroglobulin fraction, in comparison with this activity in the controls, was three- to fivefold in the bone marrow and nine- to tenfold in the spleen between days 4 and 7 after irradiation. This effect was not obtained with the isologous serum protein fraction containing proteins of smaller molecular weight.     

Maximum benefit of a precise nitrogen application system for wheat

Research is ongoing to develop sensor-based systems to determine crop nitrogen needs. To be economic and to achieve wide adoption, a sensor-based site-specific application system must be sufficiently efficient to overcome both the cost disadvantage of dry and liquid sources of nitrogen relative to applications before planting of anhydrous ammonia and possible losses if weather prevents applications during the growing season. The objective of this study is to determine the expected maximum benefit of a precision N application system for winter wheat that senses and applies N to the growing crop in the spring relative to a uniform system that applies N before planting. An estimate of the maximum benefit would be useful to provide researchers with an upper bound on the cost of delivering an economically viable precision technology. Sixty five site-years of data from two dryland winter wheat nitrogen fertility experiments at experimental stations in the Southern Plains of the U.S.A. were used to estimate the expected returns from both a conventional uniform rate anhydrous ammonia (NH₃) application system before planting and a precise topdressing system to determine the value of the latter. For prices of $0.55 and $0.33 kg⁻¹ N for urea-ammonium nitrate (UAN) and NH₃, respectively, the maximum net value of a system of precise sensor-based nitrogen application for winter wheat was about $22–$31 ha⁻¹ depending upon location and assumptions regarding the existence of a plateau. However, for prices of $1.10 and $0.66 kg⁻¹ N for UAN and NH₃, respectively, the value was approximately $33 ha⁻¹. The benefit of precise N application is sensitive to both the absolute and relative prices of UAN and NH₃.This is journal paper AEJ-260 of the Oklahoma Agricultural Experiment Station, project H-2574.