Low genetic variability in Sclerotinia sclerotiorum populations from common bean fields in Minas Gerais State,Brazil, at regional,local and micro‐scales

Sclerotinia sclerotiorum, causal agent of white mould, is the most destructive and widely distributed soilborne pathogen of common bean during the autumn–winter season in Brazil. Nevertheless, little is known about the genetic structure of the pathogen population. Microsatellite (SSR) markers and mycelial compatibility groups (MCGs) were used to characterize 118 isolates collected from 20 bean fields located in the most important growing regions of Minas Gerais State (MG). Additionally, the genetic variability among 10 isolates obtained from a single sclerotium was investigated in 10 different sclerotia. Seventy SSR haplotypes and 14 MCGs were identified among the 118 isolates. The genetic differences within bean growing areas accounted for most of the genetic variation (72%). Despite the relatively high genotypic diversity, the SSR loci were at linkage disequilibrium. Moreover, 70% of the isolates were assigned to only two MCGs, and haplotypes of a given MCG were closely related. The discriminant analysis of principal components revealed five groups. There was strong genetic differentiation between isolates collected in one municipality in southern MG when compared to other regions. Common bean resistance to white mould should be assessed with representative isolates of the five genetic groups and, if possible, of the different MCGs detected in the present study. One to five haplotypes were detected among the 10 isolates obtained from a single sclerotium. Therefore, in order to ensure genetic identity of an isolate, hyphal tip or monoascosporic isolates should be used.     

Kurilosides A1, A2, C1, D,E and F—Triterpene Glycosides from the Far Eastern Sea Cucumber Thyonidium (= Duasmodactyla) kurilensis (Levin): Structures with Unusual Non-Holostane Aglycones and Cytotoxicities

Six new monosulfated triterpene tetra-, penta- and hexaosides, namely, the kurilosides A₁ (1), A₂ (2), C₁ (3), D (4), E (5) and F (6), as well as the known earlier kuriloside A (7), having unusual non-holostane aglycones without lactone, have been isolated from the sea cucumber Thyonidium (= Duasmodactyla) kurilensis (Levin) (Cucumariidae, Dendrochirotida), collected in the Sea of Okhotsk near Onekotan Island from a depth of 100 m. Structures of the glycosides were established by 2D NMR spectroscopy and HR-ESI mass spectrometry. Kurilosides of the groups A and E contain carbohydrate moieties with a rare architecture (a pentasaccharide branched by C(4) Xyl1), differing from each other in the second monosaccharide residue (quinovose or glucose, correspondingly); kurilosides of the group C are characterized by a unique tetrasaccharide branched by a C(4) Xyl1 sugar chain; and kurilosides of the groups D and F are hexaosides differing from each other in the presence of an O-methyl group in the fourth (terminal) sugar unit. All these glycosides contain a sulfate group at C-6 of the glucose residue attached to C-4 Xyl1 and the non-holostane aglycones have a 9(11) double bond and lack γ-lactone. The cytotoxic activities of compounds 1–7 against mouse neuroblastoma Neuro 2a, normal epithelial JB-6 cells and erythrocytes were studied. Kuriloside A₁ (1) was the most active compound in the series, demonstrating strong cytotoxicity against the erythrocytes and JB-6 cells and a moderate effect against Neuro 2a cells.     

Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in Brassicaceae. It is widespread in Italy and severe outbreaks occur under conditions that favour disease development. In this study a multilocus sequence typing approach (MLST) based on the partial sequence of seven loci was applied to a selection of strains representative of the main areas of cultivation and hosts. The aim was to investigate whether the long tradition of brassica crops in Italy has influenced the evolution of different Xcc populations. All loci were polymorphic; 14 allelic profiles were identified of which 13 were unique to Italian strains. Based on the seven loci, the most common genotype within the Italian Xcc strains (AP1) was also the most representative genotype found in worldwide Xcc strains. This genotype was included in a new clonal complex in addition to three other clonal complexes already identified in Xcc populations. The phylogenetic reconstruction using a concatenated dataset of four conserved protein-coding genes, dnaK, fuyA, gyrB and rpoD, showed that the Italian strains belonged to two genetic groups. Physiological races were also investigated for the first time in Italy. The race structure of Xcc was determined by inoculating eight differential Brassica lines belonging to five species and showed that, in Italy, race 4 is the most widespread, followed by races 1 and 6. No correlation was found between allelic profiles, host of isolation, geographical origin and races, although a prevalent race was identified within the same clonal complex.     

Seroprevalence of Babesia bigemina in smallholder dairy cattle in Tanzania and associated risk factors

Variations in the seroprevalence of antibody to Babesia bigemina infection by farm and animal level risk factors were investigated for 2 contrasting regions of Tanga and Iringa in Tanzania. Tanga is situated in the eastern part of the country and has typical tropical coast climate while Iringa is situated in the Southern Highlands and has a tropical highland climate. Two hundred farms from each region were selected using simple random sampling procedure and visited once between January 1999 and April 1999. Blood samples were collected from 1329 smallholder dairy animals on selected farms for harvesting serum which was subsequently used for serodiagnosis of B. bigemina using an indirect enzyme linked immuno-sorbent assay (ELISA). Of the 1329 sera samples screened, 34.9 % were positive for B. bigemina. The prevalence was higher in Iringa Region [43%, 95% confidence intervals (CI) = 39.5-47.3] than in Tanga Region (27%, CI = 23.6-30.5). Using a logistic binomial regression model as an analytical method for predicting the likelihood of animal seropositivity, we found (in both regions) that the risk of positive reaction varied with the animal's age, history of grazing and geographical location. Seroprevalence increased with age (beta = 0.01 and 0.01 per year of age, P < 0.005 in Tanga and Iringa, respectively). Animals located in Lushoto and Iringa urban district were associated with increased risk of seropositivity [Odds ratio (OR) = 4.24, P = 0.001, for Lushoto, and OR = 1.81, P = 0.040, for Iringa Urban, respectively). Animals grazed 3 months prior to sampling had higher odds for seropositivity than zero/semi-grazed, despite farmer-reported high frequency of tick control (OR = 2.71, P = 0.0087, for Tanga, and OR = 4.53, P = 0.001, for Iringa). Our study suggests that even though herd sizes are small, B. bigemina infection is widespread in many smallholder dairy farms and endemic stability with respect to this disease has not yet been attained, but the observed levels are sufficiently high to ensure that clinical disease would be a risk.     

Green tea flavan-3-ols and oligomeric proanthocyanidins inhibit the motility of infective larvae of Teladorsagia circumcincta and Trichostrongylus colubriformis in vitro

The effects of a hot water infusion and an aqueous acetone extract of green tea (Camellia sinensis) on the motility of infective larvae of the sheep nematodes Teladorsagia circumcincta and Trichostrongylus colubriformis were investigated under in vitro conditions. The infusion and extract dose-dependently inactivated the infective larvae as assessed by the larval migration inhibition (LMI) assay. To determine the components responsible for the inhibitory activity, the hot water infusion and aqueous acetone extract of green tea were fractionated on Sephadex LH-20 and the green tea extract fractions (GTE-I-VIII) characterised by mass spectrometry. The larvae were exposed to increasing concentrations of these GTE fractions. Fractions containing epigallocatechin gallate (EGCG) and proanthocyanidin oligomers were most effective. GTE fractions were more effective against T. circumcincta than T. colubriformis larvae as assessed by the LMI assay.     

Ectopic Bone as a Nidus for Calcium Oxalate Urocystolithiasis in a Cat

A 7‐year‐old female spayed domestic shorthair cat was referred to the urology service for nonsurgical urocystolith removal. A urolith was attached to the urothelium by ectopic bone. Interventional removal without surgery was successful. Follow‐up evaluation 3 years after urolith removal revealed recurrent uroliths, bladder wall mineralization, and marked renal mineralization. This case illustrates the metaplastic potential of the urothelium and that ectopic bone should be included among the pathological factors that promote lithogenesis.     

Predicting quantitative host plant resistance against phoma black stem in sunflower

Phoma black stem is an important disease in sunflower, against which no specific management method is currently deployed in France. Relevant phenotyping methods for quantitative resistance are critical for integration of this trait into breeding programmes. Components of resistance associated with physiological resistance, and morphological traits associated with disease escape were measured on 21 sunflower genotypes under growth chamber (on seedlings), greenhouse (on adult plants), and field conditions, together with disease intensity in the field. Potential predictors were first selected for sensitivity and robustness from mixed model anova s. Analyses involving ranking tests and logistic regressions were then performed to identify predictors for field resistance. The identification of predictors for resistance involved analyses conducted in two broad steps: process‐oriented experiments, and epidemic‐oriented experiments. This stepwise approach departs from many studies aimed at identifying predictors for field resistance, which rely mainly on the computation of correlation coefficients between predictors and measured field disease variables. Predictors for quantitative resistance were identified: (i) lesion length and lesion expansion on stems of plants before flowering stage, and (ii) lesion length on first leaf petioles of seedlings. A high number of leaves and tall plants were associated with disease escape. Control genotypes for susceptibility and quantitative resistance were identified, and implications for breeding and improvements were derived from this work.