Diversity of <Emphasis Type="Italic">Colletotrichum</Emphasis> species in coffee (<Emphasis Type="Italic">Coffea arabica</Emphasis>) plantations in Mexico

The aim of this study was to identify the Colletotrichum species associated with anthracnose symptoms in coffee (Coffea arabica L.) plantations in northern Puebla, Mexico. In 2013, five surveys were conducted in different production areas and at different altitudes. Symptomatic leaves, shoots, and ripe and unripe fruits of the coffee variety Red Caturra were collected. Isolates were obtained and the Colletotrichum species were identified morphologically and characterized by multilocus sequence analyses of the ACT, CAL, GAPDH, and TUB2 genes and the rDNA region. Additionally, pathogenicity tests were conducted using six isolates. We identified C. gigasporum, C. gloeosporioides, C. karstii (two isolates), C. siamense, and C. theobromicola. This is the first report of these five species infecting leaves of coffee. The symptoms caused by these species were characterized, but the species causing Coffee Berry Disease was not found. This is the first report of a complex of species affecting coffee plants in the same geographical area in Mexico, and suggests that other complexes of species may be important pathogens in coffee-producing areas elsewhere.     

Genetic diversity evaluation of a loquat (<Emphasis Type="Italic">Eriobotrya japonica</Emphasis> (Thunb) Lindl) germplasm collection by SSRs and <Emphasis Type="Italic">S</Emphasis>-allele fragments

Loquat (Eriobotrya japonica (Thunb.) Lindl.) is a minor Rosaceae fruit of growing interest as an alternative to the main fruit crops. In this context, the selection of new cultivars to satisfy the market demand will request the suitable characterization of the available germplasm. In this work, genetic relationships among 83 loquat accessions from different countries belonging to the European loquat germplasm collection, held at the Instituto Valenciano de Investigaciones Agrarias (IVIA) in Moncada (Spain) were evaluated using microsatellites and S-allele fragments. A total of nine single sequence repeats (SSRs) from Malus and Eriobotrya genera revealed 53 informative alleles and the S-RNases consensus primers detected 11 self-incompatibility putative alleles. The combined data allow to distinguish unambiguously 80 out of the 83 accessions studied. Unweighted pair-group method (UPGMA) cluster and principal coordinates analysis (PCoA), based on Dice’s genetic distance, generally grouped genotypes according to their geographic origins and pedigrees. Discrepancies and similarities of the results obtained with other variability analysis, based on pomological traits or molecular markers, on the same loquat collection are discussed.     

Pathogenicity of two recent Western Mediterranean West Nile virus isolates in a wild bird species indigenous to Southern Europe: the red-legged partridge

ABSTRACT: West Nile virus (WNV) is an emerging zoonotic pathogen whose geographic spread and incidence in humans, horses and birds has increased significantly in recent years. WNV has long been considered a mild pathogen causing self-limiting outbreaks. This notion has changed as WNV is causing large epidemics with a high impact on human and animal health. This has been particularly noteworthy since its introduction into North America in 1999. There, native bird species have been shown to be highly susceptible to WNV infection and disease with high mortalities. For this reason, the effect of WNV infection in North American bird species has been thoroughly studied by means of experimental inoculations in controlled trials. To a lesser extent, European wild birds have been shown to be affected clinically by WNV infection. Yet experimental studies on European wild bird species are lacking. The red-legged partridge (Alectoris rufa) is a gallinaceous bird indigenous to the Iberian Peninsula, widely distributed in South Western Europe. It plays a key role in the Mediterranean ecosystem and constitutes an economically important game species. As such it is raised intensively in outdoor facilities. In this work, red-legged partridges were experimentally infected with two recent WNV isolates from the Western Mediterranean area: Morocco/2003 and Spain/2007. All inoculated birds became viremic and showed clinical disease, with mortality rates of 70% and 30%, respectively. These results show that Western Mediterranean WNV variants can be pathogenic for some European bird species, such as the red-legged partridge.     

Rectovaginal fistula with anal atresia in 5 dogs

Five dogs with rectovaginal fistula and atresia ani that had been treated by surgical correction of the malformations were studied retrospectively. Ages at presentation varied from 1 to 3 months and weight from 350 g to 7.5 kg. The histories included voiding of feces through the vulva, with or without tenesmus, usually observed after weaning. Atresia ani, presence of feces in the vaginal canal, abdominal distention, and discomfort on abdominal palpation were observed during clinical examination. Also, 3 dogs had partial tail agenesis. In all dogs, the rectovaginal fistula was isolated and transected, the vulvar and rectal defects were closed separately, and the atresia ani was repaired. Normal defecation was restored, but 1 dog had fecal incontinence that subsequently resolved. One dog died 2.5 months postoperatively, and follow-up was done on the others for periods ranging from 1.6 year to 7.7 years. Surgical correction in dogs with rectovaginal fistula and atresia ani may result in a favorable outcome, if it is done early.     

Microsatellite DNA variation within and among invasive populations of Ambrosia artemisiifolia from the southern Pannonian Plain

Although the Pannonian Plain presents one of the areas in Europe most highly infested by Ambrosia artemisiifolia, previously reported population genetic studies of the invasive species have included only a few populations from this region. Our goal was to determine the level of genetic diversity and genetic structure of A. artemisiifolia populations from the southern Pannonian Plain using microsatellite DNA markers. We documented a high level of genetic diversity within the Pannonian populations, as compared with the genetic diversity parameters previously published for the other invasive populations of A. artemisiifolia in Europe. The mean number of alleles per locus (N_A) ranged between 6.8 and 9.4, allelic richness (r) between 6.59 and 8.53 and mean number of rare alleles per locus (N_R) ranged between 3.4 and 6.4 per analysed population. A low level of among‐population differentiation was detected, with percentage of variation between populations of 3.3%, generally low pairwise F_ST values (ranged from ?0.007 to 0.152) and also according to principal coordinate analysis. A Bayesian approach revealed that most individuals did not strongly associate with any single genetic cluster, confirming a lack of genetic structuring in the analysed region. Together with the results of quantification of the level of gene flow (N_m = 5.671), these observations indicated a presence of extensive gene exchange and admixture between the populations. Therefore, A. artemisiifolia in the Pannonian Plain region has the potential for rapid expansion.     

Dissecting the genetic basis of popping ability in nuña bean,an ancient cultivar of common bean

Nuña beans (Phaseolus vulgaris L.), an ancient and pre-ceramic landrace native to Andean region, possess the property of popping and a high content of proteins and carbohydrates, which makes it an alternative nutritious snack. Knowledge on the genetic bases of popping ability is relevant for common bean improvement. Progenies derived from two nuña bean crosses were used in a generation mean based mating design to determine the inheritance and gene action for five popping related traits: length of popped seeds (PSL), popping dimension index (PDI), percentage of un-popped seeds (PUS), popping percentage average (PPA) and expansion coefficient (EC). Significant additive gene effects were found for all traits, and was the only source of the observed variation for PSL, while dominance and higher-order interactions among loci contributed to the genetic divergence for the other traits. The dominance of the cultivated over nuña alleles for PDI, PPA, EC and PUS, was confirmed by high mid-parent heterosis values and generation mean comparisons. The [d] and [dd] gene effects were in opposite direction for PPA and EC, indicating duplicate epistasis. Therefore, epistasis is likely to be an important explanation for the heterosis in both traits. For PDI and PUS, the opposite signs for [aa] and [dd] gene effects indicated that the genes for increasing popping are dispersed between the parents. Generation means and variances of BC₁P₂ indicated advantages of the backcross breeding procedure to improve the adaptation of the exotic germplasm and at the same time, transfer part of the desired donor genes to cultivated common bean. The backcross to the nuña parent could be an alternative to maintain/recover the favorable epistatic gene combinations found for PDI, PPA, EC and PUS traits.     

Differences in susceptibility to Haemophilus parasuis infection in pigs

In animal breeding programs, deoxyribonucleic acid (DNA) markers can be used to identify sires that are less susceptible to disease. These DNA markers are typically discovered in populations that display differences in susceptibility. To find those differences, it was hypothesized that sires influence their offspring responses to infection with H. parasuis. To identify differences in susceptibility, colostrum-deprived pigs derived from 6 sires were inoculated with a virulent strain of H. parasuis serovar 5. Pigs were infected at 21-d of age and euthanized 1, 2, or 3 days post-infection. Rectal temperatures, bacterial detection, clinical signs, and lesions were measured by comparing disease susceptibility in the offspring from each sire. The effect of the sire on the severity of disease in the offspring was statistically analyzed using to a 2-way ANOVA with sire and test day as fixed effects. Significant differences among sires were found for lesions, rectal temperatures from days 0-1 and 0-2 (P < 0.05) and marginal effects for clinical signs (P = 0.08). On average, the offspring of sire H94 was the most susceptible to challenge. Responses to infection were categorized to determine the clinical responses and analyzed by Chi square. Overall, 10% of all pigs infected were fully resistant to H. parasuis infection. Boar H94 didn't produce any fully resistant offspring. Differences in susceptibility to H. parasuis were observed, and the results support the hypothesis that sires influence their offspring's response to infection. Tissues from this population could be used to identify DNA markers for genetic selection of sires that produce offspring more resistant to H. parasuis infection.     

Essential Oregano Oil as a Growth Promoter for the Yellowtail Tetra,Astyanax altiparanae

This study aimed to evaluate the potential of oregano oil as a growth promoter for the yellowtail tetra, Astyanax altiparanae. The fish (1.46 ± 0.09 g) were distributed into twenty‐four 60‐L aquaria at a stocking density of 0.5 fish/L. Six isonitrogenous (350 g crude protein/kg) and isocaloric (4272 kcal gross energy/kg) diets containing 0.0, 0.5, 1.0, 1.5, 2.0 and 2.5 g/kg of oregano oil were evaluated. At the end of 90 d there was a quadratic effect of oregano oil levels for weight gain, specific growth rate, protein efficiency ratio and carcass weight, and the estimated values to maximize these variables were between 0.2 and 0.6 g/kg. There was also a quadratic effect on the feed conversion ratio, and the estimated value to improve this variable was 0.62 g/kg. A positive linear effect of the treatments was observed for muscle glycogen. With increased levels of oregano oil in the diet, there was a reduction in dry matter and ether extract and an increase in the protein content of the carcass. Thus, it can be concluded that oregano oil, at the level of 0.5 g/kg, acts as a growth promoter for A. altiparanae by improving growth performance and carcass composition.     

Nile tilapia (Oreochromis niloticus), liver morphology, CYP1A activity and thyroid hormones after Endosulfan dietary exposure

Endosulfan is a worldwide used insecticide suspected to be highly toxic to aquatic organisms, including fish. Most of the available studies have focused in water exposures, although this pollutant can be transferred through food chain. Therefore, in the present study, the effects of Endosulfan on tilapia (Oreochromis niloticus), when administered through the diet. Fish were fed 21 days with diets containing 1 and 0.5 μg g⁻¹ of Endosulfan, after which qualitative histological liver analysis showed that Endosulfan induced hepatocyte destruction, vessel endothelium rupture and increased melanomacrophages aggregates. To test lower environmentally relevant doses of Endosulfan could induce hepatic damage, as well as other negative effects, such as altered phase I metabolism and plasma thyroid hormone levels. Hence, tilapia were orally exposed to 0.1 and 0.001 μg g⁻¹ for 35 days. Low environmentally realistic doses of Endosulfan were still able to induce liver histopathological damage such as increased hepatocyte vacuolization and increased eosinophil granular cell aggregates. Liver cytochrome P450 1A activity, evaluated through ethoxyresorufin-o-deethylase (EROD), was enhanced in tilapia exposed to 0.001 μg g⁻¹, whereas the highest dose had no measurable effects in this enzyme activity. Fish exposed to 0.1 μg g⁻¹ of Endosulfan had depressed T4 plasma levels. Overall, the results of the present study further demonstrate the toxic effects of Endosulfan in tilapia when administered in the diet at environmentally relevant concentrations, which indicates that in the field food chain transfer may also be an importance source of this pollutant.     

Photoprotective potential of strawberry (Fragaria × ananassa) extract against UV-A irradiation damage on human fibroblasts

Exposure to UV-A radiation is known to induce discrete lesions in DNA and the generation of free radicals that lead to a wide array of skin diseases. Strawberry (Fragaria × ananassa) contains several polyphenols with strong antioxidant and anti-inflammatory activities. Because the major representative components of strawberry are anthocyanins, these may significantly contribute to its properties. To test this hypothesis, methanolic extracts from the Sveva cultivar were analyzed for anthocyanin content and for their ability to protect human dermal fibroblasts against UV-A radiation, as assayed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide and Comet assays. Five anthocyanin pigments were identified using high-performance liquid chromatography-diode array detection-electrospray ionization/mass spectrometry. Moreover, the strawberry extract showed a photoprotective activity in fibroblasts exposed to UV-A radiation, increasing cellular viability, and diminishing DNA damage, as compared to control cells. Overall, our data show that strawberry contains compounds that confer photoprotective activity in human cell lines and may protect skin against the adverse effects of UV-A radiation.