Unusual sialilation of three different rare genetic variants of serum DBP: Gc 1A17, Gc 1A16, and Gc 1A11

Summary The proteins of three anodal Gc1 variants, Gc 1A16, 1A11, and 1A17, are characterized by the most acidic isoelectric points observed so far among the different Gc mutants. Stepwise removal of N-acetylneuraminic acid (NANA) by treatment with neuraminidase was performed to estimate the degree of sialilation of these Gc variants. The results indicate that both proteins, the anodal and the cathodal component of these Gc 1 mutants, carry sialic acid residues. This observation is remarkable in so far as usually only the anodal component of the Gc 1 protein contains NANA and only a single residue. From the experiments carried out it can be deduced that Gc 1A16 has two NANA residues in the anodal and one NANA residue in the cathodal component. Gc 1A16 was found in four members of three generations in a Danish family; the variant segregated as a Mendelian trait. More difficult to interprete are the results obtained with the variants Gc 1A11 and Gc 1A17. Gc 1A11 probably has three NANA residues in the anodal and two NANA residues in the cathodal component. Gc 1A11 has been observed in two mother-child pairs and is presumably also a simple genetic trait. Gc 1A17 has also several NANA residues in both Gc proteins; it is suggested that the anodal component has either three or four NANA residues and the cathodal component either two or three NANA residues. Family information on this variant is not yet available.     

Distribution,causal agents,and infection dynamic of emerging ink disease of sweet chestnut in Southern Switzerland

Emerging diseases caused by both native and exotic pathogens represent a main threat to forest ecosystems worldwide. The two invasive soilborne pathogens Phytophthora cinnamomi and Phytophthora × cambivora are the causal agents of ink disease, which has been threatening Castanea sativa in Europe for several centuries and seems to be re-emerging in recent years. Here, we investigated the distribution, causal agents, and infection dynamics of ink disease in southern Switzerland. A total of 25 outbreaks were identified, 19 with only P. cinnamomi, 5 with only P. × cambivora, and 1 with both species. Dendrochronological analyses showed that the disease emerged in the last 20–30 years. Infected trees either died rapidly within 5–15 years post-infection or showed a prolonged state of general decline until death. Based on a generalized linear model, the local risk of occurrence of ink disease was increased by an S-SE aspect of the chestnut stand, the presence of a pure chestnut stand, management activities, the proximity of roads and buildings, and increasing annual mean temperature and precipitation. The genetic structure of the local P. cinnamomi population suggests independent introductions and local spread of the pathogen.     

Experimental infections with Nematodirus spathiger in rabbits.

The feasibility of infecting laboratory rabbits experimentally with the ovine nematode Nematodirus spathiger was examined. Eight-week-old rabbits were dosed either with 5000 or with 17,000 third-stage larvae and killed on days 10, 21 or 42 post-infection. With the lower dose, 20 to 40% of the inoculum were recovered at necropsy. Similar values were observed with the 17,000 dose on days 10 and 21 post-infection, but on day 42 the worm population was residual (0.6%). With both dose levels, during the course of infection, the worm populations were mainly composed of fourth-stage larvae and worm egg excretion was low. N. spathiger mainly inhabited the proximal jejunum. The results were compared with N. spathiger infection in sheep to assess the usefulness of the rabbit as an experimental model for Nematodirus infection.     

An improved breeding method for solitarious locusts

In order to unravel the physiological, endocrine, and behavioral differences between gregarious and solitarious forms of the desert locust, Schistocerca gregaria (Forsk.) (Orthoptera, Acrididae), a constant supply of rather large numbers of solitary individuals has to be guaranteed. This represents a bottleneck, mainly because of the intensity of the labor involved and limited laboratory accommodation. The method we describe here substantially reduces the space and manpower needed. The survival rate we obtained in the solitarised population was relatively high, reaching about 55%. The optimal rearing temperature proved to be 32–36 °C. Cabbage leaves and oat flakes sufficed for feeding all year round. Special racks have been designed that enable high density stacking and easy handling. The solitarisation process was monitored over ten consecutive generations. Changes in morphometrics, eye stripes, color, and behavior were recorded, of which only morphometrics, temperature related development, and mortality are discussed. A shift towards the solitarious phase was recorded, with clear differences between gregarious, 1^st generation and 7^th to 10^th generation solitarious locusts.     

The effects of concurrent experimental infections of sheep with Trichostrongylus colubriformis and T. vitrinus on nematode distributions, numbers and on pathological changes

Simultaneous infections of Trichostrongylus colubriformis and T. vitrinus in the small intestine of the sheep were examined by comparing the numbers of worms which established and their distribution within the intestine in both monospecific infections and mixed infections. The results differed depending upon the species and number of parasites. The establishment of T. colubriformis was reduced and the distribution of the nematode population was displaced posteriorly within the intestine when 30,000 larvae of both species were administered, compared with pure infections of T. colubriformis. The reduced establishment was less marked with infections of 15,000 larvae of both species and there was only a slight posterior displacement of T. colubriformis. Neither effect was evident with infections of 7,500 larvae of both species. The rate of establishment and distribution of T. vitrinus were unaffected by the presence of T. colubriformis at all three rates of infection. Atrophy of villi and hypertrophy of crypts occurred at the main site of infection in the anterior duodenum. The severity of villus atrophy was related to the number of infective larvae administered and/or the worm burden. In the ileum, beyond the main site of infection, hypertrophy of villi was only found in sheep receiving the greatest number of infective larvae.     

The biodiversity of lactic acid bacteria in Greek traditional wheat sourdoughs is reflected in both composition and metabolite formation

Lactic acid bacteria (LAB) were isolated from Greek traditional wheat sourdoughs manufactured without the addition of baker's yeast. Application of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total cell protein, randomly amplified polymorphic DNA-PCR, DNA-DNA hybridization, and 16S ribosomal DNA sequence analysis, in combination with physiological traits such as fructose fermentation and mannitol production, allowed us to classify the isolated bacteria into the species Lactobacillus sanfranciscensis, Lactobacillus brevis, Lactobacillus paralimentarius, and Weissella cibaria. This consortium seems to be unique for the Greek traditional wheat sourdoughs studied. Strains of the species W. cibaria have not been isolated from sourdoughs previously. No Lactobacillus pontis or Lactobacillus panis strains were found. An L. brevis-like isolate (ACA-DC 3411 t1) could not be identified properly and might be a new sourdough LAB species. In addition, fermentation capabilities associated with the LAB detected have been studied. During laboratory fermentations, all heterofermentative sourdough LAB strains produced lactic acid, acetic acid, and ethanol. Mannitol was produced from fructose that served as an additional electron acceptor. In addition to glucose, almost all of the LAB isolates fermented maltose, while fructose as the sole carbohydrate source was fermented by all sourdough LAB tested except L. sanfranciscensis. Two of the L. paralimentarius isolates tested did not ferment maltose; all strains were homofermentative. In the presence of both maltose and fructose in the medium, induction of hexokinase activity occurred in all sourdough LAB species mentioned above, explaining why no glucose accumulation was found extracellularly. No maltose phosphorylase activity was found either. These data produced a variable fermentation coefficient and a unique sourdough metabolite composition.     

Life cycle of Trichostrongylus retortaeformis in its natural host,the rabbit (Oryctolagus cuniculus)

The chronology of the life cycle of Trichostrongylus retortaeformis (Zeder, 1800) (Nematoda, Trichostrongyloidea) is studied in its natural host Oryctolagus cuniculus. The free living period lasted 5 days at 24 degrees C. Worm-free rabbits were each infected per os with T. retortaeformis larvae. Rabbits were killed at 12 h post-infection (p.i.) and every day from one day to 13 days p.i. By 12 h p.i., all the larvae were exsheathed and in the small intestine. The third moult occurred between 3 and 5 days p.i. and the last moult between 4 and 7 days p.i. The prepatent period lasted 12 to 13 days. The patent period lasted five and a half months. The four known life cycles of species of Trichostrongylus in ruminants were compared with that of T. retortaeformis. No significant difference was found except in the duration of the prepatent period. These similarities in the life cycles confirm the previously formulated hypotheses on the relationship between the parasites of the two host groups (Durette-Desset, 1985).     

Caspase-14 protects against epidermal UVB photodamage and water loss

Caspase-14 belongs to a conserved family of aspartate-specific proteinases. Its expression is restricted almost exclusively to the suprabasal layers of the epidermis and the hair follicles. Moreover, the proteolytic activation of caspase-14 is associated with stratum corneum formation, implicating caspase-14 in terminal keratinocyte differentiation and cornification. Here, we show that the skin of caspase-14-deficient mice was shiny and lichenified, indicating an altered stratum-corneum composition. Caspase-14-deficient epidermis contained significantly more alveolar keratohyalin F-granules, the profilaggrin stores. Accordingly, caspase-14-deficient epidermis is characterized by an altered profilaggrin processing pattern and we show that recombinant caspase-14 can directly cleave profilaggrin in vitro. Caspase-14-deficient epidermis is characterized by reduced skin-hydration levels and increased water loss. In view of the important role of filaggrin in the structure and moisturization of the skin, the knockout phenotype could be explained by an aberrant processing of filaggrin. Importantly, the skin of caspase-14-deficient mice was highly sensitive to the formation of cyclobutane pyrimidine dimers after UVB irradiation, leading to increased levels of UVB-induced apoptosis. Removal of the stratum corneum indicate that caspase-14 controls the UVB scavenging capacity of the stratum corneum.     

Diet quality and immunocompetence influence parasite load of roe deer in a fragmented landscape

The influence of landscape structure and host diet on parasite load of wildlife is still largely unknown. We studied a roe deer (Capreolus capreolus) population in a fragmented agricultural landscape in southern France to explore the relationship of gastrointestinal nematode load with spleen mass (to index immunocompetence), faecal nitrogen (to index diet quality), landscape structure and age of 33 hunt-harvested roe deer. Gastrointestinal worm counts were negatively related to faecal nitrogen and spleen mass, explaining respectively 24.7% and 9.2% of the observed variability in parasite load. Landscape structure did not appear to have a direct influence on gastrointestinal worm counts, but since animals from more open areas have a diet that is richer in nitrogen, its influence may be indirect. In conclusion, in the study area, the colonisation of the agricultural landscape does not seem to have increased the risk of gastrointestinal nematode parasitism for roe deer, possibly because access to high-quality food enhances immunocompetence.