A Controlled Safety Study of Elevated Dosages of Trimethoprim Plus Sulfadiazine in Mature Horses

A novel, proprietary, oral suspension of trimethoprim plus sulfadiazine (TMP/SDZ) had no serious adverse effects on clinical, laboratory, or pathologic parameters of mature horses when administered at up to five times the intended combined dosage of 24 mg/kg twice daily for 30 consecutive days. Loose feces was the most common observation that was likely related to TMP/SDZ treatment, and the incidence of loose feces was greater in groups treated with higher dosages of TMP/SDZ. However, all episodes were self-limiting, and no horses were treated for loose feces.     

Strategies and hurdles using DNA vaccines to fish

DNA vaccinations against fish viral diseases as IHNV at commercial level in Canada against VHSV at experimental level are both success stories. DNA vaccination strategies against many other viral diseases have, however, not yet yielded sufficient results in terms of protection. There is an obvious need to combat many other viral diseases within aquaculture where inactivated vaccines fail. There are many explanations to why DNA vaccine strategies against other viral diseases fail to induce protective immune responses in fish. These obstacles include: 1) too low immunogenicity of the transgene, 2) too low expression of the transgene that is supposed to induce protection, 3) suboptimal immune responses, and 4) too high degradation rate of the delivered plasmid DNA. There are also uncertainties with regard distribution and degradation of DNA vaccines that may have implications for safety and regulatory requirements that need to be clarified. By combining plasmid DNA with different kind of adjuvants one can increase the immunogenicity of the transgene antigen – and perhaps increase the vaccine efficacy. By using molecular adjuvants with or without in combination with targeting assemblies one may expect different responses compared with naked DNA. This includes targeting of DNA vaccines to antigen presenting cells as a central factor in improving their potencies and efficacies by means of encapsulating the DNA vaccine in certain carriers systems that may increase transgene and MHC expression. This review will focus on DNA vaccine delivery, by the use of biodegradable PLGA particles as vehicles for plasmid DNA mainly in fish.     

Thermal shock induction of triploidy in Atlantic cod (Gadus morhua L.)

The effects of thermal treatments on induction of triploidy in Atlantic cod have been investigated. Cold shock [−1.7±0.1°C at 20 min post fertilization (PF) for 2 h] was based on a previously developed protocol, and heat shocks, below the lethal threshold of 24°C, were at 16, 18 or 20°C applied 20, 30 or 40 min PF for 20 min. Cold shock did not affect larval survival and was ineffective for producing triploids (range 0–4%). A heat shock of 20°C at 20 min PF generated the highest percentages (range 66–100%) of triploid larvae at hatching, with survival ranging from 10% to 20% relative to the controls. Lower heat shock temperatures or delayed shocks increased survival but decreased the number of triploids, providing no net gain in triploid yield (range 1–9%). Heat shocks applied later than 20 min PF produced 2–4% tetraploid larvae at hatching. A thermal shock of 20°C initiated at 20 min PF and lasting 20 min proved to be the most generally efficient treatment for induction of triploidy in Atlantic cod.     

Equine hyperimmune serum protects mice against Clostridium difficile spore challenge

Clostridium (C.) difficile is a common cause of nosocomial diarrhea in horses. Vancomycin and metronidazole have been used as standard treatments but are only moderately effective, which highlights the need for a novel alternative therapy. In the current study, we prepared antiserum of equine origin against both C. difficile toxins A and B as well as whole-cell bacteria. The toxin-neutralizing activities of the antibodies were evaluated in vitro and the prophylactic effects of in vivo passive immunotherapy were demonstrated using a conventional mouse model. The data demonstrated that immunized horses generated antibodies against both toxins A and B that possessed toxin-neutralizing activity. Additionally, mice treated with the antiserum lost less weight without any sign of illness and regained weight back to a normal range more rapidly compared to the control group when challenged orally with 10⁷ C. difficile spores 1 day after serum injection. These results indicate that intravenous delivery of hyperimmune serum can protect animals from C. difficile challenge in a dose-dependent manner. Hence, immunotherapy may be a promising prophylactic strategy for preventing C. difficile infection in horses.     

Digestive capacity, intestinal morphology, and microflora of 1-year and 2-year old Atlantic cod (Gadus morhua) fed standard or bioprocessed soybean meal

The objectives of this work were to evaluate how dietary soybean meal (SBM) or a soy meal made by bioprocessing the SBM (BPSBM) to remove anti-nutritional factors affected hydrolytic capacity, amino acid absorption, intestinal morphology, and microflora along the intestinal tract of Atlantic cod at two life stages. Three fish meal based standard cod diets were formulated to contain no soy (FM control), 25% SBM, or 22% BPSBM. Prior to sampling the diets were fed to duplicate groups of 0.5 kg (1-year old) and 1.7 kg (2-year old) cod for a period of 3 months, and the groups reached 0.9 and 2.5 kg, respectively. Digesta was then sampled from different intestinal sections for analyses of trypsin and amylase activity as well as absorption of amino acids, nitrogen, and sulphur. Gastrointestinal sections were sampled for measurements of relative weight (kg^− 1 body weight), and tissues from these sections were sampled for analyses of brush border enzyme (alkaline phosphatase (ALP), leucine aminopeptidase (LAP), and maltase) activity and histological examination. Microflora was sampled from both digesta and the intestinal wall. The SBM diet stimulated relative growth of all gastrointestinal sections except the distal intestine in both age classes. Relative growth of the pyloric intestine was also stimulated by BPSBM. The pyloric caeca and the upper mid intestine were found to be the major sites for enzymatic hydrolysis of protein and starch and for amino acid absorption. Dietary SBM and BPSBM did not alter the activity of trypsin and LAP, but the activity of these enzymes in the proximal intestine was affected by age, being higher in 1-year old than in 2-year old cod. The rate of amino acid, nitrogen, and sulphur absorption along the gastrointestinal tract was not affected by SBM, but was slowed by BPSBM. Dietary SBM or BPSBM did not alter the morphology of the intestinal mucosa in any sections of the cod intestine. The distal-most structure of the intestine, a compartment with inlet and outlet (anus) valves, showed very high microbial colonisation in the mucosal brush border. Inclusion of SBM in the diet changed the intestinal microflora, increasing the population level of transient bacteria in the pyloric and mid intestine, but reducing the population level of adherent bacteria throughout the intestine. To conclude, Atlantic cod appeared to have a robust and flexible digestive system able to adjust to high dietary levels of soy protein meals.     

Introduction of genetic engineering in aquaculture: Ecological and ethical implications for science and governance

Within aquaculture, genetic engineering (GE) is emerging as a powerful method for breeding of fish and shellfish, and for developing alternative sources of feed and vaccines to combat diseases. On the other hand, the use of GE in aquaculture raises ecological, ethical and economic concerns. For instance, genetically modified (GM) feed could be spread to the aquatic environment and consumed by other marine organisms, and horizontal gene transfer may conceivably occur from DNA in feed or vaccines to a recipient genome or by faeces to the environment. Numerous reports have described beneficial effects such as viral disease resistance following DNA vaccination. However, side effects, such as activation of other genes than those which are central in immune defence mechanisms, may occur and warrant further investigations. In order to achieve sustainable introduction of GE, it is crucial that appropriate scientific investigations and ethical considerations are done prior to large-scale introduction of GE products such as DNA/GE vaccines and GM feed in commercial fish farming. This may result in a solid basis for the avoidance of potentially undesirable health and environmental effects. If GE can help make aquaculture a sustainable industry, this opens the possibility of positive market and consumer responses. This can best be achieved by involving the stakeholders from the conceptual stage to the commercial stage by facilitating a transparent process whose purpose is to inform research, to identify decision stakes, and to influence design, adoption and implementation of pro-active policy.     

Cold tolerance at three salinities in post-larval prawns,Macrobrachium rosenbergii (De Man)

Post-larval Macrobrachium rosenbergii acclimated to 27, 22, or 16°C and 5, 8, or 14 ‰ were temperature-shocked to 16°C or $\text{13°}\text{16°}\text{C}$ for 1 week. Survival was significantly better in animals acclimated to 22°C or below. Salinity had no significant effect on survival. Growth of post-larvae which had been maintained at 16°C or below for 3 weeks was followed for 1 month at 27°C; it was not significantly different from that of 27°C controls. Slower growth was observed in the 14‰ prawns. These data indicate that post-larvae stocked into cool ponds from warm hatcheries might benefit from pre-acclimation to lower temperatures. Exposure to cold does not adversely affect later growth.     

Temperature variations during grain filling obtained in growth tunnel experiments and its influence on protein content,polymer build-up and gluten viscoelastic properties in wheat

The aim of this study was to investigate effects of temperature during grain filling on gluten quality characteristics at a lower to moderate temperature range. Experiments with two wheat varieties grown in field covered by polypropylene tunnels during grain filling were performed in two seasons. Mean day temperature differences achieved within the tunnel were approximately 2–2.5 °C from the open to the closed end. There were significant effects of temperature on grain maturity time, thousand grain weight and protein content. The resistance to stretching of the gluten doughs increased with the increasing day temperatures. This was reflected in the proportion of unextractable polymeric proteins (UPP). The results suggest that increases in temperature within this temperature range affect the polymerization of polymeric proteins, giving higher molecular weights, and hence increased Rmax and stronger gluten. The two varieties differed in their response to temperature. In addition, there were seasonal variations in gluten functionality that may be associated with fluctuations in day temperatures between the seasons.     

Larval development in European hake (Merluccius merluccius L.) reared in a semi-intensive culture system

Eggs of European hake (Merluccius merluccius L.) were stripped from fish caught at sea. Larvae were kept under semi‐intensive conditions at around 12°C. In addition, eggs were incubated in single wells at 9.2, 12.7 and 14.5°C, where hatching, development and survival were closely examined. During the larval stage, a total of 299 larvae were sampled to follow development and growth. In addition a small number of juveniles were sampled. Larvae hatched approximately 4 days after fertilization, and were 2.9 mm in total length (TL). At 6‐day post hatching (dph), the larvae were 4.1 mm (TL), the jaw apparatus was developed, and the larvae had started to feed. Most of the growth during the early larval period is restricted to the head, and there is almost no increase in length for the first 3–4 weeks post hatching. Teeth and pelvic fins appear at 25 dph. Development of unpaired fins at approximately 30 dph marks the start of the larval–juvenile transition. Weaning to formulated feed was accomplished 50 dph, when external morphology was similar to that of adult hake.