High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

接种沙门氏菌苗可使新生雏鸡和强制换羽母鸡得到免疫保护

过去人们常认为,由于刚出壳雏鸡的免疫系统尚未发育成熟,同时肠道内缺乏有益的微生物,因而处于高度的敏感期,此时接种沙门氏菌苗产生的保护作用很有限。然而,最新的研究表明,在这些敏感期接种疫苗,可诱发产生保护作用。     

High yield and rapid growth of Neoparamoeba pemaquidensis in co-culture with a rainbow trout gill-derived cell line RTgill-W1

Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD.     

Possible applications of modern fish larviculture technology to ornamental fish production

There has been rapid development in the marine foodfishlarviculture technology in Europe since the early eighties,especially in the flat fish, turbot and halibut, and the bass andbream species. The most significant improvements in the eightieswere the introduction of light control, artificial reproductiontechniques, appropriate water treatment and the use of rotifersand Artemia nauplii of specific sizes and in the late eightiesand early nineties the quality enhancement of live food organismsusing specific enrichment techniques. Present research is focusedon the implementation of several microbial techniques to improvethe hygiene of live prey and fish.Many of the modern larviculture techniques being used in marine foodfish could be adapted for application in the ornamental fish industry. For examples, research in the Onamental Fish Section, Primary Production Department, Singapore has demonstrated that the use of freshwater rotifers and Artemia nauplii would enable artificial rearing of Discus in the absence of the parent fish and improve the larval performance of Gouramis and Tetra larvae. The use of such small live food organisms is likely to facilitate breeding of new fish species with small larvae. Research has also indicated that the use of diets containing vitamin C and certain immuno-stimulants improved the stress resistance of guppy. Such techniques would have important application in the fish transport, an important aspect in the ornamental fish industry     

Polyunsaturated fatty acids modulate the properties of the sex steroid binding protein in goldfish

This study examines the effects of nonesterified fatty acids on the properties of the sex steroid binding protein (SSBP) in the plasma of goldfish (Carassius auratus). Scatchard analysis revealed a single class of high affinity (Kd 1.89±0.20 nM), low capacity (Bmax 302±17 nM) binding sites for [3H]17-estradiol in female goldfish plasma. The SSBP bound 17-estradiol and testosterone with similar affinity but had much lower affinity for estriol, 17,20-dihydroxy-4-pregnen-3-one and cholesterol. Nonesterified fatty acids inhibit the binding of [3H]17-estradiol to the SSBP as a function of dose, degree of unsaturation and fatty acid chain length. Polyunsaturated fatty acids (PUFAs) such as arachidonic acid (C20:4), eicosapentaenoic acid (C20:5) and docosahexaenoic acid (C22:6) strongly inhibit the binding of [3H]17-estradiol to the SSBP. By comparison, saturated fatty acids such as heptadecanoic acid (C17:0), palmitic acid (C16:0) and stearic acid (C18:0) were without effect. Scatchard analysis and Lineweaver-Burk plots showed that PUFAs act through a competitive mechanism whereby they reduce the affinity but have no effect on the binding capacity of the SSBP. Collectively, these studies suggest that in addition to their roles as metabolic energy sources and as precursors to eicosanoids, PUFAs can be potent modulators of steroid hormone interactions with the SSBP in goldfish plasma.     

Effect of carbon/nitrogen ratio manipulation in feed supplements on Artemia production and water quality in solar salt ponds in the Mekong Delta,Vietnam

The effect of carbon/nitrogen (C/N) ratio manipulation in feed supplements on Artemia production and water quality was investigated in solar salt ponds in the Mekong Delta, Vietnam. It was assumed that development of bacterial bioflocs through C/N manipulation would improve Artemia production and water quality as demonstrated in freshwater and marine aquaculture. Twelve ponds were used for three treatments and the control, in triplicates. Green water (GW) was supplied to all ponds, with the standard Vietnamese procedure of supplying GW and chicken manure (CM) as the control (C/N 1.8). Treatment ponds were supplemented with tapioca (TAP) as carbon source, combined with either CM, pig manure (PM) or rice bran (RB), with C/N ratios of 7.4, 10.5 or 20.1 respectively. After 6 weeks of culture, no single treatment supported both improved water quality and enhanced Artemia production. Overall, improved water quality was observed at C/N 20.1 and higher Artemia production at C/N 7.4. Although external field factors could have interfered with the set‐up, this is the first study on the effect of C/N manipulation in feed supplements in Artemia pond production systems and provides the basis for development of bacterial bioflocs as a technology to improve water quality and Artemia production.     

The Pelagos Sanctuary for Mediterranean marine mammals

• 1. In February 2002, France, Italy and Monaco agreed to establish an international sanctuary for Mediterranean marine mammals. The resulting Pelagos Sanctuary encompasses over 87500 km² of the north‐western Mediterranean Sea, extending between south‐eastern France, Monaco, north‐western Italy and northern Sardinia, and surrounding Corsica and the Tuscan Archipelago.
• 2. The Pelagos Sanctuary illustrates how the tenets of Marine Protected Area (MPA) design can be reconciled with the dynamic nature of oceanic systems, because its spatial scale was defined by oceanographic and ecological considerations, specifically the location of the Ligurian permanent frontal system.
• 3. By expanding protective measures beyond national waters, the Pelagos Sanctuary also sets a precedent for the implementation of pelagic protected areas in the high seas. The Pelagos Sanctuary will contribute to the conservation of the Mediterranean Sea at two scales: (i) locally, by protecting important cetacean foraging and breeding grounds in the Ligurian Sea, and by providing ‘umbrella’ protection to other marine predators in this area; and (ii) regionally, by empowering other conservation measures, such as the Specially Protected Areas Protocol of the Barcelona Convention and the wider goals of the Agreement on the Conservation of Cetaceans of the Black and Mediterranean Seas (ACCOBAMS).
• 4. However, because few cetacean species are resident within the Sanctuary, their effective long‐term conservation will require large‐scale management and coordinated monitoring throughout the Mediterranean basin.

Copyright © 2007 John Wiley & Sons, Ltd.     

Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

Background  

Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data.