Particle Size Distribution in Two Lipid Emulsions Used for the Enrichment of Artemia nauplii as a Function of Their Preparation Method and Storage Time

The present study examined the distribution of particle sizes in two experimental standardized lipid emulsions (distributed by ICES, International Council for the Exploration of the Sea) as a function of the preparation method (hand shaking or ultrasonic blending) and as a function of storage time. A 24-h enrichment trial compared the incorporated HUFA levels in the nauplii of Artemia franciscana .
The emulsion droplets in the 50% HUFA emulsion (ICES 50, ethyl ester based) were much smaller than in the 30% HUFA emulsion (ICES 30, triacylglycerol-based) in which 90% of the droplets had a diameter below 12.3 μm as compared to 1.3 pn in ICES 50. The blending method highly affected particle sizes. High-shear blending instead of the classical hand shaking of the emulsion of both ICES 30 and 50 reduced the mean particle size from 5.06 to 1.07 μm and from 0.38 to 0.11 μm, respectively. The particle size distributions remained stable during the 1-wk storage, suggesting the absence of agglomeration or bacterial development. The fatty acid composition of 24-h enriched Artemia reflected differences in the HUFA profile of the emulsions, but was independent of observed differences in the size of the particles.     

Pilot Production of Hatchery-Reared Summer Flounder Paralichthys dentatus in a Marine Recirculating Aquaculture System: The Effects of Ration Level on Growth, Feed Conversion, and Survival

Pilot‐scale trials were conducted to evaluate growout performance of hatchery‐reared summer flounder fingerlings in a state‐of‐the‐art recirculating aquaculture system (RAS). The outdoor RAS consisted of four 4.57‐m dia × 0.69‐m deep (vol. =11.3 m³) covered, insulated tanks and associated water treatment components. Fingerlings (85.1 g mean initial weight) supplied by a commercial hatchery were stocked into two tanks at a density of 1,014 fish/tank (7.63 kg/m³). Fish were fed an extruded dry floating diet consisting of 50% protein and 12% lipid. The temperature was maintained between 20 C and 23 C and the salinity was 34 ppt. Under these conditions, growth, growth variation (CV_wt), feed utilization, and survival of fish fed to 100% and 82% of a satiation rate were compared. Due to clear changes in growth patterns during the study, data was analyzed in three phases. During phase 1 (d 1–d 196), fish showed rapid growth, reaching a mean weight of 288 g ± 105 and 316 g ± 102, with a CV_wt of 0.36 and 0.32 and FCR's of 1.38 and 1.36 in the subsatiation and satiation groups, respectively. During phase 2 (d 196–d 454), fish displayed slower growth reaching mean weights of 392 g ± 144 and 436 g ± 121, with a CV_wt of 0.37 and 0.28, and increasing FCR's of 3.45 and 3.12 in the subsatiation and satiation groups, respectively. During phase 3 (d 454–d 614), fish showed little growth reaching mean weights of 399 g ± 153 and 440 g ± 129, with a CV_wt of 0.38 and 0.29 in the subsatiation and satiation groups, respectively. Over the entire growout period (d 1–d 614), feed conversion ratios were 2.39 and 2.37 and survival was 75% and 81 % in the subsatiation and satiation treatments, respectively. The maximum biomass density reached during the study was 32.3 kg/m³. The satiation feed rate was superior to the 82% satiation rate, since it maximized growth rates, with no effect on FCR. The higher CV_wt in the subsatiation group suggests increased competition for a restricted ration led to a slower growth with more growth variation. The decrease in growth in phases 2 and 3 was probably related to a high percentage of slower growing male fish in the population and the onset of sexual maturity. This study demonstrated that under commercial scale conditions, summer flounder can be successfully grown to a marketable size in a recirculating aquaculture system. Based on these results, it is recommended that a farmer feed at a satiation rate to minimize growout time. More research is needed to maintain high growth rates through marketable sizes through all‐female production and/or inhibition of sexual maturity.     

How well does a botanical garden collection of a rare palm capture the genetic variation in a wild population?

Conservation is increasingly central to the botanic garden mission. Living plant collections are important components of conservation. Critical evaluation of living conservation collections with population genetic analysis can directly inform ex situ conservation strategy. Here, we quantify the degree of genetic variation captured through a population-based collection protocol, and explore optimal sampling for ex situ conservation. An extensive living collection derived from one population of Leucothrinax morrisii (Arecaceae) provided a model system. We compared 58 specimens from the ex situ collection with 100 individuals from throughout the parent population via 6 ISSR loci. Random bootstrapped resamples of the data were made to model differently structured ex situ collections. Mean diversity (He) differed little between the collection (0.204) and the population (0.216), and genetic distance (D) was very close (0.036). Very few private alleles were found between the collection and the population. Allelic capture, as measured by percent of private alleles, was greater than 94%. Resampled collections of different sizes captured from 48% to 94% of alleles. Pairwise comparison of bootstrapped resamples suggests that increasing the representation of half-sibling groups does not significantly increase allele capture. Increase in allele capture with increasing sample size is greatest at low resample sizes, and showed diminishing returns as resample size increased. No appreciable increase in allele capture was gained through maintaining different half-sibling groups. These data inform sampling for ex situ conservation purposes, and recommend sample sizes of at least 15 individuals, with the upper limit based on resources.     

Effects of time and concentration on mortality of phosphine-resistant Sitophilus oryzae (L) fumigated with phosphine

The effects of exposure period and phosphine concentration on mortality of susceptible and resistant Sitophilus oryzae (L) were investigated. Although S oryzae is one of the world's most serious pests of stored grain there are few data on the practical significance of phosphine resistance in this species. The strains investigated were an Australian susceptible strain, a homozygous resistant strain exhibiting a level of resistance common in Australia and an unselected field strain from China with a much stronger resistance. Fumigations were carried out at 25 degrees C on adults and mixed-age cultures. For adults of all three strains and mixed-age cultures of the susceptible and resistant Australian strains, the relationship between concentration and time could be described by equations of the form Cnt = k. In all cases n < 1, indicating that time was a more important variable than concentration. In all fumigations of adults the resistant strains were harder to kill than the susceptible strain. However, in fumigations of mixed-age cultures, which contained the tolerant pupal stage, the difference between susceptible and resistant strains was more pronounced at lower concentrations than higher concentrations. For example, at 0.02 mg litre-1 the estimated LT99.9 for mixed-age cultures of the Australian resistant strain (27 days) is 3.4 times that of the susceptible strain (8 days), but at 1 mg litre-1 there is no difference between the two strains (4 days). Limited data on the Chinese resistant strain supported this finding. Twenty-three days exposure at 0.02 mg litre-1 had no effect on mixed-age cultures of this strain, but there were no survivors after 5 days exposure to 1 mg litre-1.     

Comparison of effects of high-pressure processing and heat treatment on immunoactivity of bovine milk immunoglobulin G in enriched soymilk under equivalent microbial inactivation levels

Immunoglobulin-rich foods may provide health benefits to consumers. To extend the refrigerated shelf life of functional foods enriched with bovine immunoglobulin G (IgG), nonthermal alternatives such as high-pressure processing (HPP) may offer advantages to thermal processing for microbial reduction. To evaluate the effects of HPP on the immunoactivity of bovine IgG, a soymilk product enriched with milk protein concentrates, derived from dairy cows that were hyperimmunized with 26 human pathogens, was subjected to HPP or heat treatment. To achieve a 5 log reduction in inoculated Escherichia coli 8739, the HPP or heat treatment requirements were 345 MPa for 4 min at 30 degrees C or for 20 s at 70 degrees C, respectively. To achieve a 5 log reduction in natural flora in the enriched soymilk, the HPP or heat treatments needed were 552 MPa for 4 min at 30 degrees C or for 120 s at 78.2 degrees C, respectively. At equivalent levels for a 5 log reduction in E. coli, HPP and heat treatment caused 25% and no detectable loss in bovine IgG activity, respectively. However, at equivalent levels for a 5 log reduction in natural flora, HPP and heat resulted in 65 and 85% loss of bovine IgG activity, respectively. Results of combined pressure-thermal kinetic studies of bovine milk IgG activity were provided to determine the optimal process conditions to preserve product function.     

Genetic diversity of livestock-associated MRSA isolates obtained from piglets from farrowing until slaughter age on four farrow-to-finish farms

During a previous longitudinal study, performed on four farrow-to-finish farms (A to D), samples were taken from twelve sows, their offspring, and the environment on various occasions over six months to study the MRSA presence. During the present study, a selection of the obtained MRSA isolates were typed by multiple-locus variable-number tandem-repeat analysis (MLVA), Pulsed Field Gel Electrophoresis (PFGE), spa typing, and SCCmec typing to study the genetic diversity of LA-MRSA isolates and to determine possible MRSA sources for pig(let)s. PFGE, spa typing, and SCCmec typing revealed the presence of one or few dominant genotype(s) per farm. In contrast, 212 MLVA types were detected on the four farms, forming one cluster on farm A, three on farm B, four on farm C and two on farm D. The genotype, found on farm A was unique for this farm. Farms B, C and D shared one cluster. In general, MLVA types from these clusters were isolated from piglets, sows, and the environment on various sampling events. Piglets carried MLVA types both related and unrelated to their mother sows’ MLVA types at farrowing and onwards. In conclusion, molecular typing revealed that within a farm one or a few dominant strain(s) are widespread. Potential MRSA sources for piglets were mother sows, the environment and other piglets.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0089-4) contains supplementary material, which is available to authorized users.     

Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.