Cryptic species and population genetic structure of Plasmopara viticola in São Paulo State,Brazil

Downy mildew (Plasmopara viticola) is one of the most important diseases in grape-growing areas worldwide, including Brazil. To examine pathogen population biology and structure, P. viticola was sampled during the 2015/16 growing season from 516 lesions on nine grape cultivars in 11 locations in subtropical areas of São Paulo State, Brazil. For identification of cryptic species, a subsample of 130 isolates was subjected to cleaved amplified polymorphic sequence (CAPS) analysis, and for 91 of these isolates the ITS1 region was sequenced. These analyses suggest that the population of P. viticola in São Paulo State consists of a single cryptic species, P. viticola clade aestivalis. Seven microsatellite markers were used to determine the genetic structure of all 516 P. viticola isolates, identifying 23 alleles and 55 multilocus genotypes (MLGs). Among these MLGs, 34.5% were clonal and represented 93% of the isolates sampled. Four dominant genotypes were present in at least five different locations, corresponding to 65.7% of the isolates sampled. Genotypic diversity (Ĝ = 0.21–0.89) and clonal fraction (0.58–0.96) varied among locations (populations). Most populations showed significant deviation from Hardy–Weinberg expectations; in addition, excess of heterozygosity was verified for many loci. However, principal coordinate analysis revealed no clusters among locations and no significant isolation by distance was found, suggesting high levels of migration. The results indicate that downy mildew epidemics result from multiple clonal infections caused by a few genotypes of P. viticola, and reproduction of P. viticola in São Paulo State is predominantly asexual.     

Site‐specific management is crucial to managing Mikania micrantha

Increasingly, weeds have been taking on global distributions. With the proliferation of invasive weeds has come the challenge of managing these species over broad geographical regions, with diverse habitats and political jurisdictions. Here, we review the management of Mikania micrantha Kunth (Asteraceae; mile‐a‐minute) throughout its invaded range, extending through most of the Pacific islands and southern and south‐east Asia. Context matters when determining the best course of action for managing M. micrantha, as it has invaded a large variety of agricultural and natural systems. In Queensland, Australia and Florida, USA, M. micrantha has been targeted in relatively successful eradication campaigns, highlighting the importance of early detection and rapid response methods, while elsewhere in its invaded range, populations are either still increasing or showing limited signs of decline. An inter‐regional approach to research and management should incorporate successful management strategies employed throughout the invaded range including, but not limited to, chemical and cultural control practices, manual and mechanical control, classical biological control using the rust fungus Puccinia spegazzinii, plant–plant competition and integrated approaches utilising two or more control methods concurrently. Additional knowledge of M. micrantha genetics is required to determine if management approaches could be fine‐tuned for particular populations. Countries bordering the Mekong River formed a network in 2011 to co‐ordinate the management of invasive species such as M. micrantha. Expanding such a collaborative approach to other regions could further reduce populations of M. micrantha and limit its spread.     

Migration pathways,speed and mortality of Atlantic salmon (Salmo salar) smolts in a Scottish river and the near‐shore coastal marine environment

Long‐distance migration of Atlantic salmon (Salmo salar) is known to result in high levels of mortality. For a species experiencing global population decline, it is thus vital to better understand migration behaviour, both in the river and marine stages. Atlantic salmon smolts (n = 50) were tracked using acoustic telemetry in the River Deveron, Scotland, and adjacent coastal area. Higher rates of mortality were observed in the river (0.77% per km) than the early marine stage of migration (0.0% per km). Mortality likely resulted from predation. Higher swim speeds were recorded in the early marine stage compared with the river (marine = 7.37 ± 28.20 km/day; river = 5.03 ± 1.73 km/day [mean ± SD]), a potential predator avoidance behaviour. The majority of smolts leaving the river did so in darkness and on a flooding tide. Overall river and marine migration success were linked to nights of lower lunar brightness. Marine migration speed decreased with increasing environmental noise levels, a finding with implications for fisheries management. The migration pathway in the early marine environment did not follow obvious geographical features, such as the coastline. Thus, we suggest that early marine environment pathways are more influenced by complex water currents. These findings highlight factors that influence smolt migration survival and behaviour, areas on which future research should focus.     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish <Emphasis Type="Italic">Hoplias malabaricus</Emphasis> hepatocytes

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 μg l^?1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.     

Effect of dietary protein content on growth rate,survival and body composition of juvenile cauque river prawn,Macrobrachium americanum (Bate 1868)

We determined the effect of four concentrations of dietary crude protein, 30.7, 37.2, 41.8 and 46.8% on growth rate, survival and body composition of the juvenile cauque river prawn (Macrobrachium americanum). The prawns were hatched in the laboratory from the spawn of one wild ovigerous female. Prawns consuming 37.2% crude protein reached a final weight of 0.58 g (feed conversion ratio of 2.15), which was significantly better than the other treatments. Survival was 100% in all treatments. Protein content in the diets had no significant effect on whole body proximate composition and amino acid profile. Juveniles consuming the 37.2% crude protein diet grew faster than those fed the other diets. Specific growth rate was adjusted to the two‐slope broken‐line regression analysis model to estimate the optimal protein requirement. In conclusion, these results demonstrated that the 37.2% protein level diet is optimal for juvenile cauque river prawn M. americanum in the experimental conditions of this study.     

Understanding farmers’ potato production practices and use of improved varieties in Chencha,Ethiopia

This study was carried out to better understand non-adoption of improved varieties of potato (Solanum tuberosum) and associated technologies by smallholder farmers in Chencha, Ethiopia. Data were collected through a survey (n = 47) and in-depth interviews (n = 20). It shows how wealth status was a factor of major importance. Most wealthy and some medium-wealthy farmers adopted improved potato varieties and many of the improved production practices; they had access to seed, associated knowledge and support, and sufficient resources that were necessary to apply the improved practices. Non-adoption was common among many medium-wealthy and most poor farmers: they lacked—next to access to the technologies and knowledge—cash, land, and labor. Results indicated the need to rethink research and intervention efforts. Next to paying attention to differences in the access to technology and the related knowledge, there is a need to consider the variation in technology needs, supporting microcredit services, and room to experiment. As a result, different combinations of improved production practices may be adopted.     

Chemical composition of Lippia spp. essential oil and antimicrobial activity against Aeromonas hydrophila

Phytotherapy can replace antibiotic administration as an alternative to control Aeromonas hydrophila, one of the main bacteria involved in the aetiology of farmed fish diseases. Given that plants of the Lippia spp. genus show biological potential for antimicrobial activity, this study evaluated the chemical composition of essential oils extracted from Lippia alba, Lippia origanoides and Lippia sidoides and their activity against A. hydrophila. The oils were obtained by steam distillation in a Clevenger‐type apparatus and their composition determined by gas chromatography and mass spectrometry (CG/MS). Antibacterial activity was assessed by calculating the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using broth microdilution method. The main compounds identified were geranial (25.4%) and neral (16.6%) in L. alba oil, carvacrol (40.4%) and p‐cymene (11.4%) in L. origanoides oil and thymol (76.6%) and ortho‐cymene (6.3%) in L. sidoides oil. The three Lippia species showed bacteriostatic and bactericidal action against A. hydrophila, with MICs and MBCs ranging from 1250 to 5000 μg mL^?1. Of the species tested, the best performance was obtained with essential oil of L. sidoides.     

Enhancing capacity for freshwater conservation at the genetic level: a demonstration using three stream macroinvertebrates

1. Species diversity is declining more rapidly in freshwater ecosystems than in any other, but the consequences for genetic diversity, and hence evolutionary potential, are poorly understood. In part this reflects limited use and development of modern molecular tools and genetic approaches to address conservation questions in rivers, lakes and wetlands. As widespread, diverse and functionally important organisms, freshwater macroinvertebrates are ideal candidates for genetic approaches to reveal, for example, the conservation consequences of demographic histories and past disturbances. However, the availability of microsatellite markers for this group is very limited.
2. Using next generation sequencing, microsatellite markers were developed for Isoperla grammatica (Poda, 1761), Amphinemura sulcicollis (Stephens, 1836) and Baetis rhodani (Pictet, 1843) to enable conservation genetic investigations of these widespread invertebrate species. Fifty‐two robust microsatellite loci were developed (18, 21 and 13 per species), all with high levels of allelic diversity (7–27, 3–16, 5–13 alleles per loci, respectively).
3. These tools will allow assessment of genetic structure, dispersal and demographic resilience in these model species as a function of environmental change and variation, thereby aiding freshwater monitoring and conservation. The authors urge further capacity building to support genetic applications to the conservation biology of other aquatic organisms.

Copyright © 2016 John Wiley & Sons, Ltd.     

Impact of sampling and storage technique,and duration of storage,on the composition of fresh grass when analysed using near‐infrared reflectance spectroscopy

The objective of this study was to examine the effect of sampling technique (pluck or cut), storage duration (immediate analysis, 24‐h or 48‐h), storage temperature (ambient or chilled) and storage conditions (air present, air excluded or breathable) on the composition of fresh grass sampled from a sward managed to simulate grazing. Treatments were repeated across four sampling dates, with grass samples stored in grip seal bags prior to analysis using near‐infrared reflectance spectroscopy. Grass sampled by ‘pluck’ had a higher crude protein and ME content, and a lower acid detergent fibre (ADF) content, compared to that sampled by ‘cut’. Grass stored for 48 h had a lower water soluble carbohydrate (WSC) and ME content and a higher ADF content than for immediate analysis. Samples stored for 24 h did not differ from immediate analysis. Grass stored at ambient temperature had a lower WSC and ME content compared to immediate analysis. Grass stored under ‘breathable’ conditions had a lower ME content and higher ADF content than immediate analysis or samples stored with air present or air excluded. It is recommended that grass for analysis should be sampled by cutting, stored chilled (4°C) in a sealed bag to minimize exposure to oxygen and analysed within 24 h of harvest.