Mortality outbreak by perch rhabdovirus in European perch (Perca fluviatilis) farmed in Italy: Clinical presentation and phylogenetic analysis

This work reports a mortality outbreak, occurred in 2015 and affecting juveniles of European perch (Perca fluviatilis L.) farmed in Italy. Perch rhabdovirus (PRV) was detected by viral isolation and biomolecular investigations. Phylogenetic analysis clustered our isolate into genogroup B, which also includes PRV isolates from Perca fluviatilis identified in France (2004–2009); diagnostic investigations also revealed opportunistic bacteria (Aeromonas hydrophila) and parasites (Chilodonella piscicola). Since, occasionally, PRV has been reported in the natural environment, which is often a source of eggs and broodstock for farms, it could be possible that both similar France and Italian isolate were imported from a same place elsewhere and have a common origin. Improving biosecurity measures (batch control) and disinfection of egg strings with an iodine‐based solution helps prevent apparent vertical transmission of PRV.     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

Not the one,but the only one: about <Emphasis Type="Italic">Cannabis cryptic virus</Emphasis> in plants showing ‘hemp streak’ disease symptoms

Interveinal chlorosis and leaf margin wrinkling are widespread symptoms of Cannabis sativa. They are traditionally attributed to the so-called hemp streak virus (HSV), but its existence has not been demonstrated yet. To our knowledge, no molecular investigation has so far been performed in order to identify the causal agent of this symptomatology, we therefore decided to use traditional and molecular virology techniques to better characterize symptoms and pursue the etiological agent. No pathogenic virus was found by using targeted PCR reactions and by RNA sequencing, whereas we were able to detect the Cannabis cryptic virus (CanCV) with both techniques. We, therefore, developed an RT-qPCR assay based on a CanCV-specific TaqMan probe and applied it to a wide range of symptomatic and symptomless plants, using a two-step (for quantification), or a one-step (for fast detection) protocol. Both symptoms and the virus were only shown to be transmitted vertically and did not pass via mechanical inoculation or grafting, though we could not find any cause-effect correlation between them. In fact, the virus was found in all the tested hemp samples, and its abundance varied greatly between different accessions and individuals, independently from the presence and severity of symptoms. The suggestion that hemp streak is caused by a virus is therefore questioned. Some abiotic stresses seem to play a role in triggering the symptoms but this aspect needs further investigation. For breeding purposes, a selection of parental plants based on the absence of symptoms proved to be efficient in containment of the disease.     

Mineral contents of the muscle tissue and the digestive gland of Octopus vulgaris during short‐term starvation

We used spectrometry to study variation in the contents of minor (B, Cu, Fe, Mn and Zn) and major essential elements (Ca, K, Mg, Na and P) in the muscle tissue and digestive glands of Octopus vulgaris after 1, 2, 4 or 8 days of starvation and in fed controls. The contents of the major essential elements remained constant during 8 days of starvation. Na was the principal element found in both tissues, followed by K, P and Mg. The contents of the minor essential elements were higher in the digestive gland than in the muscle tissue, with increased contents of Fe, Mn and Zn (p < .05) during starvation, confirming the high capacity of this organ to accumulate trace minerals. In muscle tissues, B, Fe and Mn contents were lower and Cu and Zn contents higher, in starved animals than in fed animals (p < .05). Significant negative correlations between the digestive gland index and the contents of Cu, Fe and Zn suggested that the loss of weight of the digestive gland during starvation is responsible for this increase in mineral concentration. This and further information on variation in mineral content will help to optimize performance of diets formulated for the octopus.     

Feeding of yeast (Candida spp.) improves in vitro ruminal fermentation of fibrous substrates

In vitro gas production technique (IVGPT) was used with the objective of determining the inclusion effect of live cells of two strains of Candida yeast on in vitro ruminal fermentation of two fibrous substrates. In order to achieve this, two experiments were performed: A) using oat straw (Avena sativa) as substrate; B) using alfalfa hay (Medicabo sativa) as substrate, comparing the effect of two different strains of Candida genre, both isolated from the rumen, on the mentioned substrates. Levica 25 (Candida tropicalis) yeast belongs to the culture collection of the Institute of Animal Science, Cuba, and Levazoot 15 (Candida norvegensis) yeast is part of the collection of the Faculty of Zootechnology and Ecology of the Autonomous University of Chihuahua, Mexico. Both strains demonstrated their potential in activating the ruminal fermentation. They stimulated (P<0.0001) the ruminal fermentation of the substrates under study. However, the Levazoot strain stimulated the dry matter (DM) fermentation of alfalfa in 21.43%, more than Levica 25. It is concluded that there is an influence of yeast strain and diet on the rumen environment and, therefore, it is important to select the appropriate strain in every production condition.