Cloning of fox (Vulpes vulpes) Il2, Il6, Il10 and IFNgamma and analysis of their expression by quantitative RT-PCR in fox PBMC after in vitro stimulation by Concanavalin A

The immune response in the fox (Vulpes vulpes), despite the success of the oral rabies vaccine is not well characterised, and specific immunological tools are needed. A quantitative RT-PCR using SyBR Green to investigate fox cytokine expression after antigen PBMC in vitro re-stimulation is presented here. First, we cloned by homology with dog cytokine sequences the fox IL2, IL6, IL10, IFNγ and a partial 18S sequence. Fox specific primers were then defined and used to set up a species-specific quantitative RT-PCR assay using SyBR Green and 18S housekeeping gene as internal standard. The technique was validated using total RNA from fox PBMC stimulated with a polyclonal activator, Concanavaline A.     

First report of Tuta absoluta resistance to diamide insecticides

Journal of Pest Science - The tomato borer Tuta absoluta (Lepidoptera: Gelechiidae) is an invasive pest of tomato crops that is rapidly expanding around the world. It is considered a devastating...     

Energy and protein utilization in growing cattle

Limited data are available to describe the different phases of dietary protein and energy utilization in growing cattle as compared with those in adult cattle or in growing nonruminants. The European data on this topic are summarized to indicate application in appropriate feeding standards. Net protein requirements are widely variable with breed and sex. They are lower in steers than in bulls and lower in early maturing than in late maturing breeds. They are clearly defined for growing and fattening bulls where they are influenced by breed, live weight and live weight gain. New systems have been proposed to express the protein allowances. They provide a great step towards a concept explaining N supply to ruminants. However, protein degradability in the rumen, efficiency of microbial protein synthesis, intestinal digestibility and metabolic efficiency of amino acid absorption in the intestine need to be described more accurately. Even if body energy retention measured by the slaughter technique is systematically lower than when measured by calorimetric balance, both techniques can correctly describe the effect of breed sex, weight, or daily gain on energy retained, in relative value, and its distribution between protein and fat deposition. But further research is needed to confirm the distribution of metabolizable energy between maintenance and growth and the efficiency of metabolizable energy utilization for growth. Thus, different authors have preferred to calculate the energy allowances, not by a factorial method, but by regression between energy intake and the corresponding weight and daily gain of animals measured during feeding trials.     

Determination of the Antioxidant Capacity of Culinary Herbs Subjected to Various Cooking and Storage Processes Using the ABTS*+ Radical Cation Assay

Culinary herbs have the potential to be a significant source of antioxidants in the diet. However, many culinary herbs are cooked or undergo some other form of processing before they are consumed as part of a meal and such factors may affect their significance as a source of dietary antioxidants. Thus, the impact of cooking (simmering, microwaving, stewing, stir frying and grilling) and storage (vinegar maceration, cold maceration and freezing) on the antioxidant capacity of common culinary herbs was investigated. Extracts of cinnamon, cloves, fennel, ginger, lavender, parsley, rose, rosemary, sage and thyme were prepared pre and post cooking or storage and their antioxidant capacities determined using the Trolox equivalent antioxidant capacity assay (TEAC). Simmering, soup making and stewing significantly increased antioxidant capacity, whilst grilling and stir frying decreased it. Both freezing herbs at -20 degrees C and cold maceration had preservative effects on antioxidant capacity. Herbs in cold vinegar macerations for 1 week showed a decrease in antioxidant capacity compared to the control extracts. These results indicate that the potential of culinary herbs to be significant contributors to dietary antioxidant intake is significantly affected by both cooking and storage.     

Dynamics of acetylcholinesterase activity recovery in two earthworm species following exposure to ethyl-parathion

In order for cholinesterase (ChE) activity to be used as an effective biomarker in earthworms, the time course of enzyme activity inhibition and recovery must be fully characterized. A laboratory experiment was carried out using parathion as a model organophosphorus pesticide at the recommended dose (1 mg kg⁻¹) and a 10 fold higher dose (10 mg kg⁻¹), on two earthworm species (Allolobophora chlorotica and Aporrectodea caliginosa). ChE activity and weight were measured every week for a 14 day period of exposure to parathion and then for 8 weeks in uncontaminated soil. After 3 days of exposure, the weight of both earthworm species had decreased by 10–15% compared to the control, regardless of the dose used. During the remainder of the exposure period, no differences were observed between the two doses for A. chlorotica; but A. caliginosa showed rapid weight recuperation for the lowest dose applied. After 28 days and over, the control and both exposed species of worms lost similar amounts of weight. ChE inhibition was measured during and after the exposure period. ChE inhibition followed a different time course for the two species investigated. A. chlorotica appeared less sensitive to parathion than A. caliginosa. In this latter species, ChE inhibition was rapid at close to 70% of the control after 3 days, for either dose, and reached 80–90% after 7 days exposure. While A. chlorotica exhibited the same pattern of inhibition for 10 mg kg⁻¹ of parathion, the inhibition process was slower for the recommended dose with 50% inhibition after 7 days of exposure and 70% after 14 days. ChE activity recovery, after transfer to uncontaminated soil, also followed a different pattern for the two species. After exposure to 1 mg kg⁻¹ parathion, ChE activity from A. chlorotica underwent a slow but constant recovery process to regain the control value after 8 weeks in unpolluted conditions. On the other hand, the ChE activity from A. caliginosa remained strongly inhibited. The differential susceptibility to parathion found in this study could be related to differences in the specificity of the total ChE activities between those two species.     

Detection of the first lectin with antimicrobial activity present in serum of the Amazonian fish tambaqui Colossoma macropomum

Tambaqui Colossoma macropomum is the most important Amazonian native species in South American aquaculture. Innate immunity at least partially depends on the recognition of pathogen-associated molecular patterns by receptor-recognizing pathogens (PRRs). Some PRRs have been characterized in fishes, and several studies have focused on the role of lectins in the immune system of various fishes. Lectins are proteins that specifically recognize carbohydrates and which have important biological functions. Tambaqui serum lectin (ComaSeL), which was identified on the basis of its hemagglutinating activity, was pre-purified, biochemically characterized, and used in assays of antibacterial activity against pathogenic bacteria in freshwater fishes. A study of the seasonality of this lectin was performed. Comasel activity was stable at a pH between 4.0 and 9.0 and lost 100?% of its activity at 70?°C. It recognized the carbohydrates d-galactose, 1-O-methyl-d-galactopyranoside, and d-fucose, showing antibacterial activity for Gram-negative bacteria. Its activity showed significant differences between the summer and winter (p?<?0.05, Tukey test), thereby corroborating observations that tambaqui becomes more susceptible to mortality from diseases caused by bacteria and fungi during the winter. With this information, new tools may be developed for gaining a better understanding of the role of these proteins in the immune system of the tambaqui, ultimately resulting in the improved management of this fish by pisciculturists.     

Experimental approach on the selection and persistence of anti-microbial-resistant Aeromonads in faecal matter of rainbow trout during and after an oxolinic acid treatment

The presence of anti-microbial-resistant bacteria in the aquatic environment remains a matter of concern for environmental, animal and human health risks. Many published data have reported increased anti-microbial resistance among the aquatic microbiota from fish farm environments which often coincides with the use of anti-microbial agents. It can be assumed that the selection of anti-microbial-resistant bacteria first occurs in the fish farms themselves, at locations where anti-microbial concentrations are active. As anti-microbial agents are frequently administered through feed pellets, faecal matter excreted could favour conditions for such selection. In this study, OA (oxolinic acid) concentrations and OA resistance of Aeromonads were surveyed in output water and in faecal matter from OA medicated (test) and non-medicated (control) fish tanks during and after an OA oral treatment. Because of the low counts of Aeromonads in the output water, it was not possible to obtain reliable data on the proportion of anti-microbial resistance in this compartment. On the other hand, a time-limited effect on the abundance of Aeromonads isolated from faecal matter was only observed in test tanks. In these tanks, high proportion (80-100%) of OA-resistant Aeromonads was present in deposited faecal matter until at least one week after the end of the anti-microbial treatment. At that time OA was no longer detected within the faecal matter, after having reached concentrations between 100 and 190 μg/g during the treatment. We discuss how the leaching of OA from faecal pellets, from the intestine to their deposit, may favour the selection of OA-resistant Aeromonads. Further investigations in real conditions on microbiota associated with faecal matter in transit through the fish farm system are required. Particular attention should be paid to the wastes collected by the filter treatments of fish farm effluents.     

Chronic mucocutaneous candidiasis in humans with inborn errors of interleukin-17 immunity

Chronic mucocutaneous candidiasis disease (CMCD) is characterized by recurrent or persistent infections of the skin, nails, and oral and genital mucosae caused by Candida albicans and, to a lesser extent, Staphylococcus aureus, in patients with no other infectious or autoimmune manifestations. We report two genetic etiologies of CMCD: autosomal recessive deficiency in the cytokine receptor, interleukin-17 receptor A (IL-17RA), and autosomal dominant deficiency of the cytokine interleukin-17F (IL-17F). IL-17RA deficiency is complete, abolishing cellular responses to IL-17A and IL-17F homo- and heterodimers. By contrast, IL-17F deficiency is partial, with mutant IL-17F-containing homo- and heterodimers displaying impaired, but not abolished, activity. These experiments of nature indicate that human IL-17A and IL-17F are essential for mucocutaneous immunity against C. albicans, but otherwise largely redundant.