The Impact of Natural Deep Eutectic Solvents and Extraction Method on the Co-Extraction of Trace Metals from Fucus vesiculosus

In recent years, natural deep eutectic solvents (NADES) have been widely investigated for the extraction of food and medicinal plants as well as seaweeds. However, the ability of NADES for trace elements co-extraction from natural sources is not well investigated. The aim of this study was to investigate the ability of common NADES for trace elements co-extraction from Fucus vesiculosus. All of the tested NADES did not recover As and Co (concentration <LOQ). Moreover, all of the tested NADES provided a low recovery (<9%) of Ba, Ca, Fe, Mg, Mn, Sr, and Zn. The method of extraction had not shown a statistically significant effect on the co-extraction of all elements (excluding Ba and Ca). In contrast, the water content in NADES was significantly affected on the recovery of Ba, Ca, Mg, Mn, Sr, and Zn. The recovery of Al and Cr was relatively high and considerably varied (from 1.5 to 59.9%). NADES comprising lactic acid:glucose:H₂O (5:3:1) provided the lowest contents of all elements, and the highest extracted amounts were obtained employing water contents of 60–80%. The calculated daily intake of all the elements contained in NADES extracts were less than the daily dose risk estimators. The hazard quotients, hazard indexes, and carcinogenic risk calculated for all trace elements and their combination were considerably less than 1. This evidences no health risk, and carcinogenic risk after topical application of all studied NADES. For the first time, the results of the current study demonstrated that NADES extracts of F. vesiculosus contain a lower amount of trace metals and are safer than the extracts obtained with water and 70% acetone. This indicates a significant advantage for NADES compared with the other solvents.     

Breaching the black box: The role of ramps in Thai sustainable palm oil certification

Certification of sustainable palm oil as organised through the Round Table on Sustainable Palm Oil is based on a simplified understanding of the global palm oil value chain – according to which instructions about production practices can be directly translated from the palm oil mill to the primary producer. The reality of palm oil provision is much more complex than this as is shown in the case of Thailand. On the basis of qualitative field study in Southern Thailand this paper clarifies that intermediary stages, such as the collection of oil palm fruit bunches at the ramp, play a key role in the organisation of the chain. The fluidity and complexity of the palm oil flow at the local level complicates the promotion of sustainability through certification. Global and national stakeholders, such as processing and trading firms, non‐governmental organisations and national governments, should therefore open this black box of local dynamics to more effectively contribute to sustainability in palm oil supply.     

Using genetic methods for analysis of fish meals and feeds employed in Greek mariculture

Large quantities of high protein fish meals are needed to sustain cultured species and thus the impact to marine ecosystem has been highly discussed. The aim of this study was to apply a PCR‐cloning methodology for a robust insight into the composition of commercial fish meals and feeds for farmed species of the Greek mariculture, assessing the risk posed by aquaculture to marine ecosystems but also the risk posed by commercial fish feeds to the increase in trophic level of species farmed in Greece. 89% of the sequences were identified to species level and only 11% to genus/family level. Overall, a total of 49 taxa were identified (44 fish species/taxon, five non‐fish species/taxon). Even though small pelagic fish like Engraulis sp. were the main portion, a wide range of species constituted the fish meals and feeds. Plant and animal species were also detected as an alternative protein source. Feed products employed in Greek mariculture still contain large portions of fish meals which increase the mean trophic level of farmed species causing a farming up trend. The results emphasize that such molecular methodologies are needed to certify aquafeeds allowing fish feed producers to demonstrate their commitment to sustainable aquaculture.     

Subependymal Reaction Secondary to Choroid Plexus Papilloma in a Horse

Choroid plexus papilloma (CPP) is a rare, benign, intracranial neoplasm. The present report described a choroid plexus papilloma of the fourth ventricle with unusual proliferative reaction of ependymal/subependymal cells in a stallion. A single, soft neoformation, red-gray in color, with an irregular surface, resembling a cauliflower, expanding from the fourth ventricle to the cerebellar peduncles was observed. Microscopically, the neoformation was composed of a single layer of simple cuboidal to columnar epithelium with a well-vascularized papillary-arboriform pattern. Multifocal subependymal aggregates forming rosettes were observed near the mesencephalic aqueduct (MA). Glial-fibrillary-acidic-protein (GFAP) was markedly positive only in the neuropil adjacent to the proliferative areas of the MA. Ependymal/subependymal reactions secondary to CPP have never been described in equine or human neuropathology. The capabilities of ependymal cells to regenerate and eventually proliferate are controversial and unclear and, thus, require additional investigation to understand the adaptive mechanisms of this rare pathogenic condition.     

Genome-wide detection of signatures of selection in three Valdostana cattle populations

The Valdostana is a local dual purpose cattle breed developed in Italy. Three populations are recognized within this breed, based on coat colour, production level, morphology and temperament: Valdostana Red Pied (VPR), Valdostana Black Pied (VPN) and Valdostana Chestnut (VCA). Here, we investigated putative genomic regions under selection among these three populations using the Bovine 50K SNP array by combining three different statistical methods based either on allele frequencies (F_ST) or extended haplotype homozygosity (iHS and Rsb). In total, 8, 5 and 8 chromosomes harbouring 13, 13 and 16 genomic regions potentially under selection were identified by at least two approaches in VPR, VPN and VCA, respectively. Most of these candidate regions were population-specific but we found one common genomic region spanning 2.38 Mb on BTA06 which either overlaps or is located close to runs of homozygosity islands detected in the three populations. This region included inter alia two well-known genes: KDR, a well-established coat colour gene, and CLOCK, which plays a central role in positive regulation of inflammatory response and in the regulation of the mammalian circadian rhythm. The other candidate regions identified harboured genes associated mainly with milk and meat traits as well as genes involved in immune response/inflammation or associated with behavioural traits. This last category of genes was mainly identified in VCA, which is selected for fighting ability. Overall, our results provide, for the first time, a glimpse into regions of the genome targeted by selection in Valdostana cattle. Finally, this study illustrates the relevance of using multiple complementary approaches to identify genomic regions putatively under selection in livestock.     

DNA Vaccination Partially Protects Muskellunge against Viral Hemorrhagic Septicemia Virus (VHSV-IVb)

Abstract

A DNA vaccine containing the glycoprotein (G) gene of the North American viral hemorrhagic septicemia virus (VHSV) genotype IVb was developed to evaluate the immune response of fish following vaccination and evaluate its efficacy in protecting a susceptible species, the Muskellunge Esox masquinongy, against VHSV-IVb challenge. Seven weeks (539 degree-days) following vaccination with 10 μg of either pVHSivb-G or a control plasmid, Muskellunge were challenged by immersion with 10⁵ plaque-forming units (pfu)/mL of VHSV-IVb. Fish vaccinated with pVHSivb-G had a relative percent survival (RPS) of 45%. Vaccinated fish also had significantly lower mean viral titers in tissues (4.2 × 10² pfu/g) and viral prevalence (4%) than fish receiving the plasmid control vaccine (3.3 × 10⁵ pfu/g; 82%). Neutralizing antibodies were detected 28 d (308 degree-days) postchallenge (11 weeks postvaccination) in 100% of Muskellunge vaccinated with pVHSivb-G compared with only 12% of plasmid-control-vaccinated Muskellunge, suggesting robust induction of a secondary, adaptive immune response. In addition, pVHSivb-G–vaccinated Rainbow Trout Oncorhynchus mykiss challenged 7 d (100 degree-days) postvaccination with the heterologous novirhabdovirus, infectious hematopoietic necrosis virus (IHNV), experienced an RPS of 61%, compared to control fish, suggesting induction of an early and transient nonspecific antiviral immune response. This study provides an important starting point for VHSV-IVb vaccine development and useful information about the antiviral immune response elicited by DNA vaccination in a nondomesticated fish species.

Received May 1, 2016; accepted September 1, 2016     

Relationships between agronomic factors and epidemics of Phytophthora branch canker of citrus in southwestern Spain

Phytophthora branch canker, caused by Phytophthora citrophthora, has been an increasing problem in clementine (Citrus reticulata) production in Spain during last years. The disease was particularly severe in the new citrus-growing areas of the southwestern coastal areas in Huelva Province. Recent studies revealed that disease emergence was not related to either genetic drift or host specificity changes in P. citrophthora population. Therefore, the possible association of agronomic factors with the disease was investigated. A total of 110 orchards were selected arbitrarily from the main citrus-growing areas in Huelva Province. The presence of branch cankers together with agronomic factors including soils, cultivars, rootstocks, irrigation, pruning, techniques to improve fruit production, fungicide treatments, presence of brown rot of fruit and frost damage were recorded. Logistic regression analysis was used to detect correlations between the agronomic factors studied and disease prevalence. Phytophthora branch canker was significantly associated with mature clementine orchards. Sweet orange and hybrid cultivars as well as young clementine orchards were less affected by the disease. Although disease was less frequent in Salorthid soils, alternative high resolution procedures are required to draw conclusions about the effect of soil properties on disease prevalence. As in other Phytophthora-induced diseases, soil flooding during the rainy season was correlated positively with the prevalence of branch cankers. Improving fruit production by branch scoring showed a strong positive correlation with Phytophthora branch canker. This is the first time that girdling has been associated with Phytophthora disease epidemics on a fruit tree crop, but further research is needed to determine the cause of this relationship. Cultural practices including pruning, regulated deficit irrigation, additional phosphonate sprays, and abiotic and disease factors such as frost damage and presence of brown rot of fruit were not significantly correlated with disease prevalence.     

Phytophthora boehmeriae boll rot: a new threat to cotton cultivation in the mediterranean region

A boll rot of cotton (Gossypium hirsutum L.) was observed for the first time in Greece in August 1993 in Larissa and Volos counties, and in August and September 1995 in Trikala and Phthiotis counties. Fungi of the genusPhytophthora were isolated from diseased plants. Morphological characteristics of the pathogen were recorded on mounts made directly from the infected tissues or after growth of the isolated fungus on corn meal agar or sterile distilled water. Colony morphology, growth rates, features of asexual and sexual structures and maximum growth temperatures were examined. APhytophthora species new to Europe,Phytophthora boehmeriae Sawada, attacking cotton bolls, was identified. The pathogenicity of the isolates was confirmed by artificial inoculations of detached cotton bolls. Analysis of α-esterase isozymes revealed unique banding patterns for isolates ofP. boehmeriae compared with those ofP. cactorum andP. parasitica, which arePhytophthora species with similar morphology.     

Collar rot caused byPhytophthora citrophthora on pear trees in Greece

A severe crown rot of pear trees of cultivar ‘Kondoula’ grafted on quince rootstock was observed in Greece. Isolations from the affected tissues repeatadly yielded aPhytophthora sp. that was determined by morphological and physiological characteristics to beP. citrophthora. The pathogenicity of two of theP. citrophthora isolates was tested by inoculating trunks of 2-year-old pear trees by mycelial agar disks. Thirty-two days after inoculation all inoculated trees were infected. Although the pear isolates could not be differentiated from isolates ofP. palmivora orP. nicotianae based on isozyme profiles of α-esterase or lactate dehydrogenase, RAPD profiles with one selected primer differentiated the pear isolates from the other species and revealed an electrophoretic banding pattern similar to that of aP. citrophthora standard. This is the first report ofP. citrophthora on pear trees in Greece.