Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach

With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.     

Comparative Characterization of Enzymatic Digestion from Fish and Soybean Meal from Simulated Digestive Process of Pacific Bluefin Tuna,Thunnus orientalis

The digestive process of the Pacific bluefin tuna (PBT), Thunnus orientalis, was simulated through two phases of in vitro digestion: acidic digestion with porcine pepsin, followed by alkaline digestion with pancreatic crude extract (PCE) obtained from the PBT to hydrolyze fish meal (FM) and soybean meal (SBM) as protein substrates. The crude protein from FM resulted in a lower degree of hydrolysis (73.3%) compared with SBM (79.2%). However, the resulting digested products showed that FM contained 35% more small peptides, with sizes <6.5 kDa than those from the starting material (>150 kDa). The SBM had an increase of only 1.3% in the similar peptide cut‐offs found after hydrolysis. These results suggested that FM appeared to be a better source of protein according to the amount of low‐molecular weight peptides. In addition, the proteolytic activity of PCE showed that 88.9% of its alkaline proteolytic activity corresponded to trypsin and 2.9% corresponded to chymotrypsin activity. The results shown here demonstrate that peptide sizes are important in identifying suitable protein sources for aquafeed production to reinforce the primary results obtained from the in vitro digestibility using the pH‐Stat system. These results also contribute to a better understanding of the digestibility process in aquatic organisms.     

Associations of fish with various types of littoral habitats in reservoirs

Fish associations with different types of littoral habitats were studied in four canyon‐shaped reservoirs in the Czech Republic in years 2010 and 2011 by gillnets. Two to three habitats per reservoir–beaches (former meadows), stump fields (former forest) and rubble slopes–were defined and sampled along the longitudinal axis of reservoirs. Effects of reservoir, habitat and locality (position along longitudinal axis) on fish biomass, abundance and species structure were tested for juvenile and adult fish separately. Hierarchical analysis of variance revealed that habitats differed significantly in fish biomass and abundance. Redundancy analysis showed that analysed environmental variables had significant influence on fish community structure. Most variability in community structure was explained by reservoir and then by combination of habitat and slope steepness. Locality position had the smallest influence on community structure. For both adult and juvenile fish total abundance and biomass, the most inhabited habitat was beaches; rubble slopes were the least inhabited. Habitat associations differed among species. Among adults, bream Abramis brama, white bream Blicca bjoerkna and roach Rutilus rutilus were associated with beaches and stump fields, whereas perch Perca fluviatilis, ruffe Gymnocephalus cernuus, asp Aspius aspius and pike Esox lucius were associated with rubble slopes. Bream, white bream, bleak Alburnus alburnus, roach, ruffe and pikeperch Sander lucioperca were associated with beaches among juveniles, whereas the only juvenile associated with rubble slopes was perch. We showed that most common species are associated with distinct habitats and also that utilisation of various littoral habitats differs in general.     

Angiographic Anatomy of External Iliac Arteries in the Sheep

External iliac artery atherosclerotic disease and aneurism occur in man. For treatment, imaging is required to facilitate minimally invasive introduction and advancement of stents within the intended vessels. Sheep are commonly used to test and improve stents. However, little information is published regarding the angiographic anatomy of the iliac arteries in the ovine species. The objective of this study was to describe the angiographic anatomy of the iliac arteries in the sheep. Computed tomography (CT) angiography and gross anatomical dissection were performed in, respectively, 10 and 43 adult ewes. Diameters and lengths of the arteries were measured. In comparison with man, salient anatomical differences were identified in the sheep: (1) the absence of common iliac arteries, (2) the common trunk at the origin of internal iliac arteries and (3) the location of the bifurcation of the external iliac arteries into femoral arteries in the pelvis (not in the limb). External iliac arteries in this series of sheep were 86 mm long in average and had a mean diameter of 7.5 mm. Lengths of arteries are only slightly different between man and sheep, while diameters are rather similar. Therefore, the sheep model appears to be sufficiently similar to man to test stent properties. This study provides useful reference images and measures of lengths and diameters of relevant arteries that could be applied to research with ovine models.     

Evaluation of the effectiveness of two infectious bronchitis virus vaccine programs for preventing disease caused by a California IBV field isolate

Infectious bronchitis virus CA99 serotype was isolated from several broiler flocks in Northern California. The virus caused late-onset respiratory disease and increased airsacculitis condemnation in affected flocks despite the use of an established infectious bronchitis virus vaccination program. An experimental study compared Holland/Arkansas and Massachusetts/Arkansas vaccination protocols to determine the efficacy of commercial infectious bronchitis virus vaccines in reducing respiratory disease and airsacculitis lesions found at processing that were associated with a CA99 field isolate. All vaccination groups were given Massachusetts/Connecticut strains of infectious bronchitis virus vaccines at age 1 day followed by vaccination with either Holland/ Arkansas or Massachusetts/Arkansas vaccine strains at 18 days of age. Birds were challenged at age 31 days with a CA99 field isolate. Gross pathology, histopathology, and virus isolation were evaluated. Chickens vaccinated with Holland/Arkansas had marginally better protection against CA99 challenge than chickens vaccinated with Massachusetts/Arkansas, although differences were not statistically significant.