Genetic diversity and pathogenic variability among isolates of colletotrichum species from strawberry

ABSTRACT Ninety-five isolates of Colletotrichum including 81 isolates of C. acutatum (62 from strawberry) and 14 isolates of C. gloeosporioides (13 from strawberry) were characterized by various molecular methods and pathogenicity tests. Results based on random amplified polymorphic DNA (RAPD) polymorphism and internal transcribed spacer (ITS) 2 sequence data provided clear genetic evidence of two subgroups in C. acutatum. The first subgroup, characterized as CA-clonal, included only isolates from strawberry and exhibited identical RAPD patterns and nearly identical ITS2 sequence analysis. A larger genetic group, CA-variable, included isolates from various hosts and exhibited variable RAPD patterns and divergent ITS2 sequence analysis. Within the C. acutatum population isolated from strawberry, the CA-clonal group is prevalent in Europe (54 isolates of 62). A subset of European C. acutatum isolates isolated from strawberry and representing the CA-clonal and CA-variable groups was assigned to two pathogenicity groups. No correlation could be drawn between genetic and pathogenicity groups. On the basis of molecular data, it is proposed that the CA-clonal subgroup contains closely related, highly virulent C. acutatum isolates that may have developed host specialization to strawberry. C. gloeosporioides isolates from Europe, which were rarely observed were either slightly or nonpathogenic on strawberry. The absence of correlation between genetic polymorphism and geographical origin in Colletotrichum spp. suggests a worldwide dissemination of isolates, probably through international plant exchanges.     

Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis.

Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR analysis and histologic examination suggested a leptospiral infection in the newborn foal. At the moment of death, the infection coexisted with a streptococcal-associated aspiration bronchopneumonia and postpartum septicemia. These findings indicate that the PCR assay based on the amplification of the hap1 gene represents a useful tool for specific detection of pathogenic leptospira in field samples taken from horses.     

Production of meiotic gynogenetic and triploid sea bass, Dicentrarchus labrax L.: 1. Performances, maturation and carcass quality

Meiotic gynogenetic and triploid sea bass were produced by pressure shocks according to a previously published protocol. Pressure-treated groups did not survive as well as controls during early development and larval rearing. Performances, sexual maturation and carcass quality were examined over a period of 34–45 months. At the age of 34 months, growth of the gynogenetic fish was comparable to that of the control but inferior in the triploid fish. A predominance of male fish was found within the triploid groups, while diploid and meiotic gynogenetic fish showed equal proportions of the sexes. Gonadal maturation in triploid fish was significantly impaired, particularly in the females that showed rudimentary ovaries. Triploid males exhibited primary maturation but proved to be gametically sterile. Pressure-induced triploids did not grow as well as diploids, but these results might be ascribed to specific on-growing conditions (communal rearing). The performance of gynogenetic sea bass was comparable to that of control. The superiority of diploid fish over their triploid counterparts was confirmed during the final growing period and more clearly so in females. Performances of triploids varied according to their maternal origin. Overall, striking qualitative differences between diploid and triploid fish were found at the age of 34 and 45 months, although the results varied in a gender-specific manner. A strong maternal effect was also observed. The potential advantages of triploid sea bass for aquaculture purposes are discussed.     

A faster inoculation assay for Armillaria using herbaceous plants

Armillaria (honey fungus) is a virulent necrotrophic pathogen that causes Armillaria root disease. Conventional Armillaria inoculation assays use young saplings as hosts and consequently are cumbersome, frequently conducted outdoors, and take many years from establishment to analysis of infection. We have developed and evaluated a faster inoculation assay for Armillaria that uses herbaceous plants as hosts, is carried out in controlled conditions, and reduces experimental durations to 3 months. Plant species of known susceptibility to Armillaria and comparisons between virulent A. mellea and opportunistic A. gallica were used to validate the assay. Mortality and diagnostic symptoms of Armillaria root disease such as epiphytic rhizomorphs and mycelial fans were used to assess levels of infection. We also attempted to reduce assay preparation time by substituting woody inocula with agar inocula, but typical symptoms of Armillaria root disease were only observed on plants infected with woody inocula. Through our assay, we identified five new potential herbaceous hosts of Armillaria: Kniphofia hirsuta, Hordeum vulgare, Lobelia cardinalis, Nicotiana tabacum and Helenium hoopesii – further expanding the extensive list of plants with susceptibility to Armillaria and suggesting infection of herbaceous species may be more widespread than currently acknowledged.     

Electron microscopic observation of the respiratory tract of SPF piglets inoculated with Mycoplasma hyopneumoniae.

Seven hysterectomy derived piglets were repeatedly challenged with Mycoplasma hyoneumoniae during the first week of life. Samples of trachea, bronchi and lung tissue collected 2–11 weeks post-inoculation (p.i.) were examined using light and electron microscopy. Autoradiography was used to study in more detail the site of M. hyopneumoniae multiplication. Gross lesions were observed in lung tissue and were characterized by hyperplasia of the epithelium and an increased mononuclear cell accumulation in perivascular and peribronchiolar areas. Mild lesions of the trachea and the bronchi, including epithelial hyperplasia and infiltration of the lamina propria by inflammatory cells, were noted.

Electron microscopy showed that, 2–6 weeks p.i., changes in the mid-trachea and bronchi surface consisted of the loss of cilia. Mycoplasmas covered tufts of cilia remaining on the epithelial cell surface. Scanning and transmission electron micrographs showed that they were predominantly found closely associated with the top of cilia. No specialized terminal structure could be seen and no mycoplasma cells were identified lying free in the lumen nor in close contact with the plasma membrane of cells of microvilli. Some fine fibrils radiating from one mycoplasma to another or to cilia were seen at higher magnification by scanning electron microscopy. Six to eleven weeks p.i., a disrupted epithelial surface lacking cilia was observed. Cells were desquamated and shed into the lumen with cellular remains containing droplets of mucus.

Autoradiography revealed that label corresponded to the observed mycoplasma distribution. At the top of cilia, a high density of labeling was visible in the zone of high mycoplasma concentration. Therefore, incorporation of the label in the mycoplasma is proof or their multiplication in the trachea.

The intimate association between the mycoplasma and cilia may be an important factor in the pathogenesis of the disease caused by M. hyopneumoniae (swine enzootic pneumonia).     

Effects of ketanserin on hypergranulation tissue formation, infection, and healing of equine lower limb wounds

The objective of this multicentre, randomized, controlled field study was to determine the efficacy of ketanserin gel in preventing exuberant granulation tissue formation (hypergranulation) and infection in equine lower limb wounds. Horses and ponies (n = 481) with naturally occurring wounds were randomized to either topical treatment with ketanserin gel (n = 242) or a positive control (Belgium, Germany: ethacridin lactate solution, n = 120; France, United Kingdom: malic, benzoic, and salicylic acid [MBS] cream, n = 119). Treatment continued until the wound healed (success), formed hypergranulation tissue (failure), or became infected (failure). Treatment was terminated after 6 months in all remaining animals. Ketanserin was successful in 88% of cases. Wounds treated with ketanserin were 2 and 5 times more likely to heal successfully than were those treated with MBS or ethacridin lactate, respectively. Ketanserin gel is thus more effective than these standard treatments in preventing hypergranulation tissue and infection of equine lower limb wounds.     

Influence of thermal processing conditions on acrylamide generation and browning in a potato model system

Fried potato products such as French fries and chips may contain substantial amounts of acrylamide. Numerous efforts are undertaken to minimize the acrylamide content of these products while sensory properties such as color and flavor have to be respected as well. An optimization of the frying process can be achieved if the basic kinetic data of the browning and acrylamide formation are known. Therefore, heating experiments with potato powder were performed under controlled conditions (moisture, temperature, and time). Browning and acrylamide content both increased with heating time at all temperatures and moisture contents tested. The moisture content had a strong influence on the activation energy of browning and acrylamide formation. The activation energy strongly increased at moisture contents below 20%. At higher moisture contents, it was very similar for both parameters. At low moisture contents, the activation energy of acrylamide formation was larger as compared to the one for browning. This explains why the end of the frying process is very critical. Therefore, a lower temperature toward the end of frying reduces the acrylamide content of the product while color development is still good.     

Effect of Geogenic Lead on Fungal and Collembolan Communities in Garden Topsoil

Geogenic lead (Pb) is considered to be less bioavailable than anthropogenic Pb and exerts less effect on the soil fauna. However, Pb contamination in vegetables has been reported in the case of geogenic anomalies, even at moderate concentrations (around 170 mg kg^–1). In this study, we investigated collembolan communities using both taxonomic- and trait-based approaches and observed fungal communities to assess the effects of a moderate geogenic Pb anomaly on collembolans and fungi in an urban vegetable garden soil. Results indicated that geogenic Pb indeed modified fungi communities and altered the functional structure of collembolan communities in garden soils. Although geogenic Pb presented low bioavailability, it affected soil fauna and vegetables similar to anthropogenic Pb.     

Spatial distribution of sediments and transfer properties in soils in a stormwater infiltration basin

Purpose  

The aim of our study was to characterise the heterogeneity of sediment distribution in a stormwater retention/infiltration basin (Pont de Cheviré, Nantes, France) and to determine the impact of this distribution on water transfer properties in the soil.     

Potential of Aquatic Macrophytes as Bioindicators of Heavy Metal Pollution in Urban Stormwater Runoff

The concentrations of heavy metals in water, sediments, soil, roots, and shoots of five aquatic macrophytes species (Oenanthe sp., Juncus sp., Typha sp., Callitriche sp.1, and Callitriche sp.2) collected from a detention pond receiving stormwater runoff coming from a highway were measured to ascertain whether plants organs are characterized by differential accumulations and to evaluate the potential of the plant species as bioindicators of heavy metal pollution in urban stormwater runoff. Heavy metals considered for water and sediment analysis were Cd, Cr, Cu, Ni, Pb, Zn, and As. Heavy metals considered for plant and soil analysis were Cd, Ni, and Zn. The metal concentrations in water, sediments, plants, and corresponding soil showed that the studied site is contaminated by heavy metals, probably due to the road traffic. Results also showed that plant roots had higher metal content than aboveground tissues. The floating plants displayed higher metal accumulation than the three other rooted plants. Heavy metal concentrations measured in the organs of the rooted plants increased when metal concentrations measured in the soil increased. The highest metal bioconcentration factors (BCF) were obtained for cadmium and nickel accumulation by Typha sp. (BCF = 1.3 and 0.8, respectively) and zinc accumulation by Juncus sp. (BCF = 4.8). Our results underline the potential use of such plant species for heavy metal biomonitoring in water, sediments, and soil.