Fungal endophytes in grasses from semi-arid permanent grasslands of western Spain

Abstract A survey of grass species associated with Epichloë and Neotyphodium fungal endophytes was made in permanent semi‐arid grasslands of western Spain. Endophytes were isolated from 11 of 49 grass species analysed. Infected plants of Festuca arundinacea, Festuca rubra, Dactylis glomerata and Lolium perenne were detected at most locations studied. The grasses, Agrostis castellana, Brachypodium phoenicoides and Festuca ampla, are previously unknown endophyte hosts. The results obtained show that endophyte–grass interactions are present in the grasslands where this study was conducted.     

Evidence on the response of Patagonian forage grasses to the mulching effect of recent tephra deposits in Argentina

In Patagonia, arid and semiarid lands are being affected both by inappropriate management practices, which are leading to degradation, and by volcanic activity, whose effects are still unclear. This study aimed to test whether superficial deposition of volcanic tephra could benefit two of the most prominent Patagonian forage grass species (Poa ligularis and Pappostipa speciosa var. speciosa). Pots with P. ligularis and P. speciosa were kept under wet (W) and dry (D) conditions in the presence (T+) or absence (T?) of tephra for 105 days, and then were all well-watered. We determined the effects of tephra on soil water retention and conservation, soil moisture content (% v/v), plant growth, stomatal conductance (gs), and gs recovery capacity. The water regime significantly affected both species performance and gs. The presence of tephra increased soil water conservation, soil moisture content in wet conditions, and P. ligularis gs in wet conditions, and decreased senescence in dry conditions (9% in P. ligularis and 16% in P. speciosa). The presence of tephra allowed roots to grow in 8/10 and 2/10 pots in W conditions for P. ligularis and P. speciosa, respectively, and in only 1/10 pots in D conditions, only for P. ligularis. Tephra was also associated with gs recovery after dry conditions. Poa ligularis was more positively affected by tephra than P. speciosa, probably because P. ligularis has higher phenotypic plasticity. The positive effects of tephra may increase the resilience and resistance of P. speciosa and P. ligularis to periods of water shortage.     

Physiological and population genetic analysis of Botrytis field isolates from vineyards in Castilla y León,Spain

Grey mould is reported in the vineyards of Castilla y León, Spain, every year. However, the natural populations of the pathogen have yet to be properly characterized. Vineyards from six wine-producing areas were surveyed in 2002 and 2007, sampling from bunches of grapes with and without symptoms. A total of 283 Botrytis field isolates were selected for physiological and genetic analyses. Botrytis cinerea isolates predominated in the population, although isolates belonging to Botrytis pseudocinerea and Botrytis prunorum were also identified. These two species are recorded for the first time in Spain in this work. In addition, two isolates closely related to Botrytis californica were identified. Physiologically, the B. cinerea population is very diverse, displaying a normal distribution of aggressiveness values in Vitis vinifera leaves, suggesting a quantitative nature for this trait. Several isolates unable to cause infection were identified, most of them belonging to a mycelial morphotype. Population genetic analysis revealed that genotypic diversity is high and that multiple infections of the same bunch of grapes by different genotypes occur frequently. The high genotypic diversity observed, an even distribution of both mating types and the linkage disequilibrium values detected support a mixed mode of reproduction with low levels of clonality. The wine-producing area in which each isolate was collected imposed a low degree of population differentiation, an effect that does not depend solely on the geographic distances but rather on the management practices used by growers and wine producer associations.     

The infection of Festuca rubra subsp. pruinosa by Epichloë festucae

Festuca rubra subsp. pruinosa is a grass that inhabits the sea cliffs of the Atlantic coasts of Europe. Four populations of this plant in the northern coast of Galicia, in northern Spain, were analysed for the presence of fungal endophytes. Of the plants included in the study, on average 0·69 were infected by an intercellular fungal endophyte. Using morphological and molecular characters, this endophyte was identified as Epichloë festucae, a clavicipitaceous, seed‐transmitted species. A half‐sibling line, consisting of endophyte‐free and endophyte‐infected plants, was developed and used to test whether the presence of the fungus affected the response of the plants to salinity. Dry weights of infected or uninfected plants, watered with sea‐water solutions or tap water, did not significantly differ.     

Seroprevalence of Sarcocystis neurona and Its Association With Neurologic Disorders in Argentinean Horses

Equine protozoal myeloencephalitis (EPM) is generally caused by Sarcocystis neurona and can produce substantial economic losses on equine production in America. The aims of the present study were to evaluate the seroprevalence of S. neurona in the main horse-production area of Argentina and associate it with the occurrence of neurologic disorders. Serum samples were collected from 640 horses in nine Argentinean provinces. Most of the samples correspond to animals ≥1.5-year-old from different breeds (n = 628); 12 samples were from younger horses. Further seroprevalence comparison was conducted from the older animals grouped with (n = 148) or without neurologic signs (n = 480). Immunoblot: proteins from 2 × 10⁷S. neurona merozoites were used as antigen on each membrane. Reactivity to antigens with relative mobility of 7, 10, and 16 kDa was considered specific for antibodies against S. neurona; reactivity at 30 kDa was recorded separately. The overall seroprevalence for S. neurona was 26.1% (167/640), and all the provinces had positive horses. Seroprevalence of animals with neurologic signs was greater (P < .001) than what was observed in normal horses (39.2% vs. 22.1%), with an odds ratio of 2.27. Reactivity at 30 kDa was detected in 71% of all samples. This study identified a wide distribution of S. neurona–positive animals in Argentina and horses with neurologic signs having a greater seroprevalence than normal horses. Sarcocystis neurona infection should be considered for early differential diagnosis and treatment of animals with neurologic disorders to decrease the economic impact of EPM in Argentina.     

Standardization and validation of an agar gel immunodiffusion test for the diagnosis of equine infectious anemia using a recombinant p26 antigen

We developed and validated an agar gel immunodiffusion test (AGID) test for the diagnosis of equine infectious anemia (EIA) using as antigen the p26 protein of equine infectious anemia virus (EIAV) produced in the Escherichia coli expression system. The developed rp26-AGID test showed an excellent diagnostic relative sensitivity (100%) and specificity (100%) compared to a commercial AGID assay when 1855 field serum samples were analyzed. In addition, the rp26-AGID demonstrated to be a precise assay with excellent repeatability and reproducibility. In the analytical sensitivity trial, positive sera showed nearly the same endpoint dilutions for both compared tests. No positive-reactions were observed with 35 serum samples with antibodies related to other endemic agents and also with severely hemolysed samples, demonstrating that the rp26-AGID has an excellent analytical specificity. Complete concordance with blind previous results from five proficiency test panels confirmed the capability of the assay of accurate detection of EIAV antibodies. This is the first time that a recombinant AGID assay able to identify EIAV infections has been standardized and validated in Argentina according to international guidelines. Taking into account the results obtained, the p26-AGID could be adopted as an official test method for the diagnosis and control of EIA in this country.     

Effects of choke disease in the grass Brachypodium phoenicoides

Using phenotypic and molecular characters, as well as mating compatibility tests, the fungus causing choke disease in Brachypodium phoenicoides was identified as Epichloë typhina . A three year field experiment conducted with infected and uninfected plants of a single clone of B. phoenicoides showed no significant differences in biomass production during their vegetative growth stage, but the content of calcium, magnesium and manganese was significantly greater, and that of sodium was significantly lower in infected plants compared to uninfected plants. Infected plants produced up to twice as many reproductive tillers as healthy plants, but their reproductive tissue biomass was significantly smaller than that of healthy plants, because tiller development was arrested by choke forming stromata.     

Anatomy and Histology of the Male Reproductive Tract and Spermatogenesis Fine Structure in the Lesser Anteater (Tamandua tetradactyla,Myrmecophagidae, Xenarthra): Morphological Evidences of Reproductive Functions

The anatomy and histology of the male genital tract of the lesser anteater were studied. Fine details of spermatozoa regarding their genesis and morphology were also studied in six adult specimens. The testes lie in the pelvic cavity. The deferent duct emerges from the epididymis and opens into the ejaculatory duct, which drains into the membranous urethra. Accessory glands (prostate, seminal vesicle and bulbourethral gland) are histologically similar to those described in other mammals. The short penis presents an urethral orifice, while the corpus spongiosum becomes thinner at the end indicating the absence of a histologically defined glans. The seminiferous epithelium shows: (1) Sertoli cells with deep nuclear indentations, (2) spermatogonia with crusty‐like chromatin, (3) spermatocytes at different stages of maturation and (4) three morphologically distinct stages of spermatid differentiation according to nuclear shape, acrosome development and chromatin condensation. Sperm heads appear oval. The length of the spermatozoa averages 67.33 ± 1.60 μm. Two specimens with inactive spermatogenesis were azoospermic. Their testes and epididymis presented sizes smaller than those with active spermatogenesis. These studies together with others in anteaters may contribute to successful breeding in conservation programmes.     

The infection of Festuca rubra by the fungal endophyte Epichloë festucae in Mediterranean permanent grasslands

The purpose of this work was to determine the frequency of infection by fungal endophytes in Festuca rubra plants from Mediterranean permanent grasslands (dehesas). Plants infected with the fungal endophyte Epichloë festucae were detected in all twenty-seven populations studied in the province of Salamanca, Spain. Infection frequencies in six selected populations ranged from 44% to 92% (average, 70%) of the plants. Infected plants were almost always symptomless. These results suggest that the association between E. festucae and F. rubra is common and occurs at a high frequency in Mediterranean permanent grasslands.     

Movements,behavior, and habitat utilization of yellowfin tuna (Thunnus albacares) in waters surrounding the Revillagigedo Islands Archipelago Biosphere Reserve,Mexico

The movements, behavior, and habitat utilization of yellowfin tuna, Thunnus albacares, following capture and release with archival tags in the Revillagigedo Islands Archipelago Biosphere Reserve (RIABR), Mexico, are described from analyses of 16 578 days of time‐series data, downloaded from 52 archival tags recovered from yellowfin (78–173 cm in length and 1.7–8.0 yr of age) at liberty from 93 to 1773 days ( = 411 days), collected during 2006–2012. An unscented Kalman filter model with sea‐surface temperature measurements integrated (UKFsst) was used to process the archival tag data sets to obtain improved estimates of geographic positions, most probable tracks (MPTs), and movement parameters. The MPTs indicate restricted movements, low levels of dispersion, and fidelity of yellowfin tuna to the RIABR. The median parameter estimates from the UKFsst model for errors in longitude (σ_x) and latitude (σ_y) were 0.46° and 1.84°, respectively, for directed movements (u and v) –0.05 NM day^?1 and –0.05 NM day^?1, respectively, and for dispersive movement (D) 117.99 NM² day^?1. Analyses of daily timed depth and temperature records resulted in the classification of the data into four distinct behaviors. There are significant differences among ages in the durations of Type I and Type II diving behaviors and in the daytime and nighttime vertical habitat utilization distributions. The oceanography surrounding the RIABR appears to have a profound influence on the movements, behavior, and habitat utilization of yellowfin in this area.