Batrachochytrium dendrobatidis is a fungus belonging to the Phylum Chytridiomycota, Class Chytridiomycetes, Order Chytridiales, and is the highly infectious aetiological agent responsible for a potentially fatal disease, chytridiomycosis, which is currently decimating many of the world's amphibian populations. The fungus infects 2 amphibian orders (Anura and Caudata), 14 families and at least 200 species and is responsible for at least 1 species extinction. Whilst the origin of the agent and routes of transmission are being debated, it has been recognised that successful management of the disease will require effective sampling regimes and detection assays. We have developed a range of unique sampling protocols together with diagnostic assays for the detection of B. dendrobatidis in both living and deceased tadpoles and adults. Here, we formally present our data and discuss them in respect to assay sensitivity, specificity, repeatability and reproducibility. We suggest that compliance with the recommended protocols will avoid the generation of spurious results, thereby providing the international scientific and regulatory community with a set of validated procedures which will assist in the successful management of chytridiomycosis in the future.

/pdf/diagnostic-assays-and-sampling-protocols-for-the-detection-2gocrwtm5f.pdf

Diagnostic assays and sampling protocols for the detection of Batrachochytrium dendrobatidis

In the last decade betanodavirus infections have emerged as major constraints on the culture of marine fish in all parts of the world with the exception of the African continent. The occurrence of these infections appears to be a function of the number of species cultured and the intensity of culture. This has been further complicated by the promiscuous translocation of stock within and between countries. Great strides have been made in defining these agents and producing diagnostic techniques but much more remains to be done. Lack of knowledge of the epidemiology of the diseases caused by nodaviruses, except for vertical transmission of the pathogen in some species, has impeded the development of control measures but, even so, the measures identified to date have not been adequately implemented by producers with the result that catastrophic losses still occur on a regular basis.

Betanodavirus infections of teleost fish: a review

The outbreaks of viral encephalopathy and retinopathy (VER), caused by nervous necrosis virus (NNV), represent one of the main infectious threats for marine aquaculture worldwide. Since the first description of the disease at the end of the 1980s, a considerable amount of research has gone into understanding the mechanisms involved in fish infection, developing reliable diagnostic methods, and control measures, and several comprehensive reviews have been published to date. This review focuses on host–virus interaction and epidemiological aspects, comprising viral distribution and transmission as well as the continuously increasing host range (177 susceptible marine species and epizootic outbreaks reported in 62 of them), with special emphasis on genotypes and the effect of global warming on NNV infection, but also including the latest findings in the NNV life cycle and virulence as well as diagnostic methods and VER disease control.

Betanodavirus and VER Disease: A 30-year Research Review.

Viral encephalopathy and retinopathy (VER), otherwise known as viral nervous necrosis (VNN), is a major devastating threat for aquatic animals. Betanodaviruses have been isolated in at least 70 aquatic animal species in marine and in freshwater environments throughout the world, with the notable exception of South America. In this review, the main features of betanodavirus, including its diversity, its distribution and its transmission modes in fish, are firstly presented. Then, the existing diagnosis and detection methods, as well as the different control procedures of this disease, are reviewed. Finally, the potential of selective breeding, including both conventional and genomic selection, as an opportunity to obtain resistant commercial populations, is examined.

/pdf/viral-encephalopathy-and-retinopathy-in-aquaculture-a-review-3lhm7tau86.pdf

Viral encephalopathy and retinopathy in aquaculture: a review

http://digital.csic.es/bitstream/10261/49429/1/Nodavirus%20increases.pdf

Nodavirus increases the expression of Mx and inflammatory cytokines in fish brain.

In a series of three 17 d replicate trials, a total of 36 ileal cannulated pigs were used to determine the effects of volatile fatty acid (VFA)inclusion in drinking water on ileal microflora, hemolytic Escherichia coli, VFA concentrations, pH, dry matter (DM), and pig performance. All pigs were weaned at 21 d-of-age and assigned to one of three treatments including: Tl)ad libitimi access to water, T2)ad libitum access to water containing acetate(50mM), propionate(5mM), and butyrate (3mM), and T3)ad libitum access to water containing acetate(25mM), propionate(2.5mM), and butyrate(1.5mM). Individual water intake was measured daily and feed intake was measured twice weekly. All pigs were individually caged in an environmentally controlled room with ad libitiam access to a phase starter diet. Ileal samples were collected at 21, 24, 28, 31, 35, and 38 d-ofage. At 38 d-of-age, pigs were sacrificed and contents were collected from the stomach, duodenum, ileum, cecum, and spiral colon. Digesta were analyzed for total E. coli, streptococci, lactobacilli, VFA, pH, and DM. Neither water nor feed intake differed between treatments and no differences were observed in microflora concentrations

The effect of volatile fatty acid inclusion in drinking water on intestinal microflora, VFA, pH, dry matter, and performance in weanling pigs

. Earthworm invasion in North American forests has the potential to greatly impact soil microbial communities by altering soil physicochemical properties, including structure, pH, nutrient availability, and soil organic matter (SOM) dynamics. While most research on the topic has been carried out in northern temperate forests, little is known on the impact of invasive earthworms on soil microbial communities in the boreal forest, a region characterized by a slower decay of organic 15 matter (OM). Earthworm activities can increase OM mineralization, altering nutrient cycling and biological activity in a biome where low carbon (C) and nitrogen (N) availability is typically limiting microbial and plant growth. Here, we characterized and compared microbial communities of earthworm-invaded and non-invaded soils in previously described sites across three major soil types found in the Canadian boreal forest using a space-for-time approach. Microbial communities of forest floors and surface mineral soils were characterized using phospholipid fatty acid (PLFA) analysis and metabarcoding of the 16S 20 rRNA gene, for bacteria and archaea, and ITS2 region for fungi. In forest floors, the effects of earthworm invasion were minor. In mineral soil horizons, earthworm invasion was associated with higher fungal biomass and greater relative abundance of ectomycorrhizal fungi. Oligotrophic bacteria (Acidobacteriota and Chloroflexi) were less abundant in invaded mineral soils, where Gram(+):Gram(

Earthworm-invaded boreal forest soils harbour distinct microbial communities

The coat protein encoded by the nodavirus RNA2 gene originally isolated from greasy grouper, Epinephelus tauvina, was cloned, expressed as a recombinant polyhistidine-tailed fusion protein and characterized by immunoblot analysis. The purified recombinant protein was used to develop an indirect enzyme-linked immunosorbent assay (ELISA) to detect body exudate and plasma antibodies against the coat protein in both experimentally infected and commercial barramundi. In addition, the nucleotide sequence was employed to develop a RT-PCR detection assay based on the T4 region. The results showed that the virus could be detected as early as 3 days post-infection by RT-PCR while antibodies against the recombinant coat protein were detectable on day 6 post-infection. Among 112 commercial barramundi samples collected from October 1999 to April 2000, 9% showed positive ELISA results which were further verified by Western blot.

Detection of nodavirus in barramundi, Lates calcarifer (Bloch), using recombinant coat protein‐based ELISA and RT–PCR

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The effect of volatile fatty acid inclusion in drinking water on intestinal microflora, VFA, pH, dry matter, and performance in weanling pigs

Earthworm-invaded boreal forest soils harbour distinct microbial communities

Detection of nodavirus in barramundi, Lates calcarifer (Bloch), using recombinant coat protein‐based ELISA and RT–PCR