Haematogenous spread of Sporothrix schenckii in cats with naturally acquired sporotrichosis

The recovery of Sporothrix schenckii from blood samples is rare, and the diagnosis of systemic sporotrichosis is usually made at necropsy. In this report, S schenckii was isolated from two or more internal organs of nine necropsied cats with naturally acquired sporotrichosis. Haematogenous spread was demonstrated in vivo by the isolation of S schenckii from the peripheral blood of 17 (n = 49, 34.4 per cent) cats. Feline leukaemia virus (FeLV) was not detected, and co-infection with feline immunodeficiency virus (FIV), observed in nine cases (n = 43, 20.9 per cent), apparently did not affect the isolation of S schenckii from peripheral blood or from the internal organs.     

Uncovering the Bioactive Potential of a Cyanobacterial Natural Products Library Aided by Untargeted Metabolomics

The Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC) holds a vast number of cyanobacteria whose chemical richness is still largely unknown. To expedite its bioactivity screening we developed a natural products library. Sixty strains and four environmental samples were chromatographed, using a semiautomatic HPLC system, yielding 512 fractions that were tested for their cytotoxic activity against 2D and 3D models of human colon carcinoma (HCT 116), and non-cancerous cell line hCMEC/D3. Six fractions showed high cytotoxicity against 2D and 3D cell models (group A), and six other fractions were selected by their effects on 3D cells (group B). The metabolome of each group was organized and characterized using the MolNetEnhancer workflow, and its processing with MetaboAnalyst allowed discrimination of the mass features with the highest fold change, and thus the ones that might be bioactive. Of those, mass features without precedented identification were mostly found in group A, indicating seven possible novel bioactive molecules, alongside in silico putative annotation of five cytotoxic compounds. Manual dereplication of group B tentatively identified nine pheophytin and pheophorbide derivatives. Our approach enabled the selection of 7 out of 60 cyanobacterial strains for anticancer drug discovery, providing new data concerning the chemical composition of these cyanobacteria.     

General and specific combining ability in Serrasalmidae

This work was carried out to predict the combining abilities, both general and specific, for performance traits and bodily yields of Serrasalmidae. Ninety‐six 30‐day‐old juveniles were purchased from two commercial fish farms, 12 of each of the following eight genetic groups: pacu, pirapitinga, tambaqui, tambacu, tambatinga, patinga, paqui and piraqui. Six fish from each genetic group were grown in 500‐L fibreglass tanks (two tanks per genetic group) until they were 495 days old. At the end of the growth period fish were weighed, subjected to morphometric analysis and processed to obtain their bodily yields. Two nuclear markers and one mitochondrial marker were used to confirm the identity of the animals. Combining abilities were obtained using the method proposed by Griffing in 1956 (Australian Journal of Biological Science, 4, 463–493) adapted to a mixed models analysis, environmental effects were estimated by the empirical best linear unbiased estimator method and genetic effects (general and specific combining abilities) were estimated with the empirical best linear unbiased predictor. Predictions of the combining abilities of advanced hybrids were obtained by the mixed models mixture method with normal distributions. Tambaqui showed higher general and specific combining abilities than the other groups for most of the variables, making it the most important genetic group. General combining ability makes a greater contribution to phenotypic variance than specific combining ability for most variables, indicating a predominance of genes with an additive effect in the control of evaluated traits.     

Use of salmon by‐product meals as a replacement for anchovy meal in practical diets for the Pacific white shrimp (Litopenaeus vannamei)

A series of trials were conducted with Pacific white shrimp Litopenaeus vannamei, to evaluate the efficiency of two salmon meals as compared to anchovy meal. The basal diet contained 200 g/kg anchovy meal, which was systematically replaced (0%, 50%, 75% and 100%) with salmon meal on an isonitrogenous basis. Another two diets were formulated with a hydrolysed salmon meal to replace 50% and 100% anchovy meal. Each diet was randomly fed to four replicate groups of 25 and 30 shrimp per tank in clear (indoor) and green (outdoor) water trials, respectively. The results showed that growth performance and feed conversion ratio were not statistically different when salmon meal replaced anchovy meal in both trials. However, when hydrolysed salmon meal was used to replace 100% of the anchovy meal, growth performance of the shrimp significantly decreased. The four kinds of fish meal (anchovy, salmon by‐product meals and menhaden) were evaluated in an ingredient digestibility trial using the 70:30 replacement technique. In general, dry matter, energy, protein and individual amino acids digestibility of salmon meal were significantly higher than those of menhaden and anchovy meal. Results of this study demonstrated that salmon meals are a good protein sources which can replace anchovy meal.     

Extraction and Characterization of Collagen from Elasmobranch Byproducts for Potential Biomaterial Use

With the worldwide increase of fisheries, fish wastes have had a similar increase, alternatively they can be seen as a source of novel substances for the improvement of society’s wellbeing. Elasmobranchs are a subclass fished in high amounts, with some species being mainly bycatch. They possess an endoskeleton composed mainly by cartilage, from which chondroitin sulfate is currently obtained. Their use as a viable source for extraction of type II collagen has been hypothesized with the envisaging of a biomedical application, namely in biomaterials production. In the present work, raw cartilage from shark (Prionace glauca) and ray (Zeachara chilensis and Bathyraja brachyurops) was obtained from a fish processing company and submitted to acidic and enzymatic extractions, to produce acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC). From all the extractions, P. glauca PSC had the highest yield (3.5%), followed by ray ASC (0.92%), ray PSC (0.50%), and P. glauca ASC (0.15%). All the extracts showed similar properties, with the SDS-PAGE profiles being compatible with the presence of both type I and type II collagens. Moreover, the collagen extracts exhibited the competence to maintain their conformation at human basal temperature, presenting a denaturation temperature higher than 37 °C. Hydrogels were produced using P. glauca PSC combined with shark chondroitin sulfate, with the objective of mimicking the human cartilage extracellular matrix. These hydrogels were cohesive and structurally-stable at 37 °C, with rheological measurements exhibiting a conformation of an elastic solid when submitted to shear strain with a frequency up to 4 Hz. This work revealed a sustainable strategy for the valorization of fisheries’ by-products, within the concept of a circular economy, consisting of the use of P. glauca, Z. chilensis, and B. brachyurops cartilage for the extraction of collagen, which would be further employed in the development of hydrogels as a proof of concept of its biotechnological potential, ultimately envisaging its use in marine biomaterials to regenerate damaged cartilaginous tissues.     

Quantitation Overcoming Matrix Effects of Lipophilic Toxins in Mytilus galloprovincialis by Liquid Chromatography-Full Scan High Resolution Mass Spectrometry Analysis (LC-HR-MS)

The analysis of marine lipophilic toxins in shellfish products still represents a challenging task due to the complexity and diversity of the sample matrix. Liquid chromatography coupled with mass spectrometry (LC-MS) is the technique of choice for accurate quantitative measurements in complex samples. By combining unambiguous identification with the high selectivity of tandem MS, it provides the required high sensitivity and specificity. However, LC-MS is prone to matrix effects (ME) that need to be evaluated during the development and validation of methods. Furthermore, the large sample-to-sample variability, even between samples of the same species and geographic origin, needs a procedure to evaluate and control ME continuously. Here, we analyzed the toxins okadaic acid (OA), dinophysistoxins (DTX-1 and DTX-2), pectenotoxin (PTX-2), yessotoxin (YTX) and azaspiracid-1 (AZA-1). Samples were mussels (Mytilus galloprovincialis), both fresh and processed, and a toxin-free mussel reference material. We developed an accurate mass-extracted ion chromatogram (AM-XIC) based quantitation method using an Orbitrap instrument, evaluated the ME for different types and extracts of mussel samples, characterized the main compounds co-eluting with the targeted molecules and quantified toxins in samples by following a standard addition method (SAM). An AM-XIC based quantitation of lipophilic toxins in mussel samples using high resolution and accuracy full scan profiles (LC-HR-MS) is a good alternative to multi reaction monitoring (MRM) for instruments with HR capabilities. ME depend on the starting sample matrix and the sample preparation. ME are particularly strong for OA and related toxins, showing values below 50% for fresh mussel samples. Results for other toxins (AZA-1, YTX and PTX-2) are between 75% and 110%. ME in unknown matrices can be evaluated by comparing their full scan LC-HR-MS profiles with those of known samples with known ME. ME can be corrected by following SAM with AM-XIC quantitation if necessary.     

Follicle Development during Luteal Phase and Altrenogest Treatment in Pigs

Synchronization of the oestrous cycle of gilts using altrenogest treatment has been found to increase ovulation rate. The current experiment investigated if the increase in ovulation rate after altrenogest treatment is related to increased follicle size at the end of altrenogest treatment compared with late luteal phase follicles. Crossbred gilts (n = 15) received altrenogest during 18 days [20 mg Regumate (Janssen Animal Health, Beerse, Belgium)], starting 5-7 days after onset of first oestrus. Control gilts (n = 15) did not receive altrenogest. At days 10-12 of the oestrous cycle [i.e. in the presence of corpora lutea (CL)], average follicle development was 2.51 +/- 0.20 mm (assessed with ultrasound) in altrenogest-treated gilts and 2.58 +/- 0.16 mm in control gilts (p > 0.10). During the last days of altrenogest treatment (i.e. when CL had gone into regression), average follicle size had increased to 3.01 +/- 0.31 mm (p < 0.05). Subsequent ovulation rate was 16.6 +/- 1.7 in altrenogest treated gilts and 15.1 +/- 1.2 in control gilts (p < 0.05). Altrenogest treatment resulted in increased follicle size after regression of the CL, showing that suppression of follicle growth by altrenogest alone is less severe than suppression by endogenous progesterone (either with or without altrenogest). Altrenogest treatment also resulted in a higher ovulation rate. However, it is unclear if the increased follicle size and higher ovulation rate after altrenogest treatment are causally related, as the relation between the two on an animal level was not significant.     

Cytotoxic tumor necrosis factor activity produced by equine alveolar macrophages: preliminary characterization

Blood monocytes and alveolar macrophages (AM) were harvested from foals (aged 46 days to 6 months) and cultured in either medium alone or medium containing 10 micrograms/ml bacterial lipopolysaccharide (LPS). After 24 h, culture supernates were collected and analyzed for cytotoxic activity on sensitized L929 cells. Both monocytes and AM that had been treated with LPS produced significantly more cytotoxic activity than the same cell type exposed to medium lacking LPS. LPS-treated macrophages secreted significantly more cytotoxic activity (120 +/- 17.8 U/ml) than did LPS-treated monocytes (47.3 +/- 17.0 U/ml); however, constitutive production of cytotoxin by monocytes was higher (16.7 +/- 4.1 versus 1.2 +/- 1.2 U/ml). The identification of the cytotoxin as tumor necrosis factor (TNF) was strongly suggested by its reactivity with a rabbit antiserum directed against the N-terminal 15 amino acids of human TNF. TNF secretion by AM increased in a dose-dependent manner between LPS concentrations of 0.0001 and 1 microgram/ml, then leveled off. Most of the cytotoxic TNF activity produced by AM was secreted within the first 8 h after initial contact with LPS. Macrophage supernatant TNF was stable over a pH range of 6-11, but lost activity when kept at a pH less than 6. Equine TNF also was destroyed by exposure for 1 h to temperatures more than 60 degrees C. TNF bioactivity was recovered as a single peak after crude macrophage supernate was subjected to analysis by either anion exchange or gel filtration chromatography (molecular weight approximately 56,000).