Genetic structure of Xiphinema pachtaicum and X. index populations based on mitochondrial DNA variation

The dagger nematodes Xiphinema pachtaicum and X. index are two of the most widespread and frequently occurring Xiphinema spp. co-infesting vineyards and other crops and natural habitats worldwide. Sexual reproduction is rare in these species. The primary objective of this study was to determine the genetic structure of X. pachtaicum and X. index populations using eight and seven populations, respectively, from different "wine of denomination of origin (D.O.) zones" in Spain and Sardinia (Italy), by studying mitochondrial (cytochrome oxidase c subunit 1 or COI) and nuclear (D2-D3 expansion segments of 28S rDNA) markers. Both Xiphinema spp. showed low intraspecific divergence among COI sequences, ranging from 0.2% (1 base substitution) to 2.3% (10 substitutions) in X. pachtaicum and from 0.2% (1 base substitution) to 0.4% (2 substitutions) in X. index. Population genetic structure was strong for both species. Nevertheless, molecular differences among grapevine-growing areas were not significant, and intrapopulation diversity was very low. It is hypothesized that this genetic homogeneity in the nematode populations reflects their predominant parthenogenetic reproduction mode and low dispersal abilities. Our results also show that X. pachtaicum populations in Spain have possibly been established from two different populations of origin. Results also demonstrated that the two DNA regions studied are suitable diagnostic markers for X. index and X. pachtaicum.     

Exploring soil bacterial communities in different peanut-cropping sequences using multiple molecular approaches

Soil bacterial communities have significant influence on soilborne plant pathogens and, thus, crop health. The present study focuses on ribotyping soil bacterial communities in different peanut-cropping sequences in Alabama. The objective was to identify changes in microbial assemblages in response to cropping sequences that can play a role in managing soilborne plant pathogens in peanut. Four peanut-cropping sequences were sampled at the Wiregrass Research Station, Headland, AL in 2006 and 2007, including continuous peanut, 4 years of bahiagrass followed by peanut, peanut-cotton, and peanut-corn-cotton. Soil sampling was done at early and mid-season and at harvest. Bacterial community structure was assessed using ribosomal intergenic spacer analysis (RISA) combined with 16S rRNA cloning and sequencing. RISA results indicated >70% dissimilarities among different cropping sequences. However, 90% similarities were noticed among replicated plots of the same cropping sequences. Cropping sequences and time of soil sampling had considerable effect on soil microbial community structure. Bahiagrass rotation with peanut was found to have the highest bacterial diversity, as indicated by a high Shannon Weaver Diversity index. Overall, higher bacterial diversity was observed with bahiagrass and corn rotations compared with continuous peanut. The bacterial divisions Proteobacteria, Acidobacteria, Firmicutes, Bacteroidetes, and Actinomycetes were the predominant bacterial phyla found in all peanut-cropping sequences. The Proteobacteria taxa in these soils were negatively correlated with the abundance of members of division Firmicutes but, conversely, had a significant positive correlation with Gemmatimonadetes taxa. The prevalence of the division Actinomycetes was negatively correlated with the relative abundance of members of division Verrucomicrobia. These results indicate complex interactions among soil bacteria that are important contributors to crop health.     

Prevalence of mixed trematode infections in an abattoir receiving cattle from northern Portugal and north-west Spain

Between February 2007 and January 2009, a total of 776 slaughtered animals were examined for the presence of Trematoda in the liver, gall bladder, oesophagus and stomach (rumen and reticulum). Data collected were analysed by the age and breed of the animals. The percentage of cattle from which Trematoda were found was 38 per cent (95 per cent confidence interval [CI] 35 to 41 per cent); 28 per cent (95 per cent CI 25 to 31 per cent) had Fasciola hepatica, 12 per cent (95 per cent CI 10 to 14 per cent) Calicophoron daubneyi and 6 per cent (95 per cent CI 4 to 8 per cent) Dicrocoelium species. A significantly high prevalence of fasciolosis and dicrocoeliosis was observed in cattle over 10 years of age. Autochthonous Rubia Gallega cattle had the highest prevalence of fasciolosis and crossbred cattle had the highest prevalence of dicrocoeliosis (P<0.05). Twenty per cent (95 per cent CI 15 to 25 per cent) of the cattle positive for Fasciola also had Calicophoron species; 10 per cent (95 per cent CI 6 to 14 per cent) also had small liver flukes (Dicrocoelium species).     

Ability of the fungus Duddingtonia flagrans to adapt to the cyathostomin egg-output by spreading chlamydospores

The analysis of the capability of the nematode trapping-fungus Duddingtonia flagrans to adapt to the cyathostomin egg-output in horses was evaluated. Fecal samples from 196 pasturing autochthonous Pura Raza Galega horses were collected from the rectum and then divided according to the egg-output into three groups: ≤ 300, 310-800 and >800 eggs per gram feces. Four doses of chlamydospores (0.1, 0.2, 0.4 and 0.8 × 10(6)/100g feces) were directly spread onto fecal pats on the ground, remaining one without treatment as control. Fecal pats confirmed the presence of gastrointestinal nematode larvae belonging to strongylid cyathostomins (Cyathostomum and Gyalocephalus spp). An overall 94% (95% CI 91, 97) percentage of reduction was obtained, and an increase in the activity of the trapping-fungi simultaneously to the rising in the number of cyathostomin eggs and larvae in the coprocultures was detected. A significantly highest reduction of the cyathostomin L3 in the coprocultures with more than 800 EPG was found, which indicates that Df trapping activity is larvae nematode density-dependant. The present research showed the high biological activity of D. flagrans against nematode larvae can adjust to the cyathostomin egg-output, and underlines its efficacy as a practical method for the control of these parasites in grazing horses.     

Brachylaima aspersae n. sp. (Digenea: Brachylaimidae) infecting farmed snails in NW Spain: morphology, life cycle, pathology, and implications for heliciculture

The life cycle of Brachylaima aspersae n. sp. (Trematoda: Brachylaimidae) in heliciculture farms is elucidated in light of field and experimental studies. Embryonated asymmetrical eggs (33.3 μm × 20.2 μm) are passed in the faeces of the definitive host, the domestic mouse (Mus musculus), and are ingested by its unique first intermediate host, the helicid snail Helix aspersa aspersa. After hatching, the miracidium develops into a highly branched sporocyst in the connective tissues of the digestive gland. Microcaudate cercariae emerging from this gastropod migrate up the ureter of the second intermediate host, the snails H. a. aspersa and H. a. maxima, and develop into non-encysted metacercariae in the kidney. Following predation of infected snails, the metacercariae develop into adults preferentially in the proximal portion of the duodenum of the definitive host. The strict oioxenic character for the first intermediate host, as well as the cercarial chaetotaxy (3 C(I)V+1 C(I)D, 10 C(II), 5 C(III)V, 14 C(III)L, 2 C(III)D, 16 H, 6 S(I), 6 S(II), 6 S(III), 2 A(I)L+1 A(I)V, 1 A(II)L, 3 ML, 1 P(I)L and 3 P(III)L), the distinct pars prostatica, the variable appearance of testes (rounded to irregular, with smooth or slightly to moderately lobulated margins), the size of eggs, the position of acetabulum (located somewhat posterior to the anterior third of body), and the microhabitat of the adult in the final host allow differentiation of B. aspersae from other well-known species in the genus. Massive infections with sporocysts or metacercariae of this brachylaimid may induce extensive pathological changes in the organs affected. Our results confirm that control of rodents in heliciculture farms is essential to minimize the potential health risks and morbimortality associated with this newly described species.     

Diagnosis of Eimeria species using traditional and molecular methods in field studies

The objective of this study was to identify and characterize species of Eimeria in broiler chickens using traditional morphological and pathological plus molecular (DNA amplification) diagnostic methodologies. Using a combination of those techniques it was possible to identify the presence of multiple circulating species in the flock as well as higher frequencies for some of them, especially Eimeria praecox and Eimeria maxima, which were identified in 100% of the flocks. The frequencies of the other species were Eimeria mitis and Eimeria necatrix (93.3%), Eimeria tenella (76,7%), Eimeria acervulina (56.7%) and Eimeria brunetti (16.7%). However using the lesion score, the most common species were E. maxima (46.7%), E. acervulina (30%), E. tenella (23.3%), and E. necatrix (10%). E. brunetti and E. praecox were not identified by using lesion score. DNA amplification had detection sensitivity for Eimeria species in the field samples of at least 20 oocysts. The implementation of DNA amplification as a routine diagnostic technique in aviaries can assist Eimeria population.