Issues and special features of animal health research

ABSTRACT: In the rapidly changing context of research on animal health, INRA launched a collective discussion on the challenges facing the field, its distinguishing features, and synergies with biomedical research. As has been declared forcibly by the heads of WHO, FAO and OIE, the challenges facing animal health, beyond diseases transmissible to humans, are critically important and involve food security, agriculture economics, and the ensemble of economic activities associated with agriculture. There are in addition issues related to public health (zoonoses, xenobiotics, antimicrobial resistance), the environment, and animal welfare.Animal health research is distinguished by particular methodologies and scientific questions that stem from the specific biological features of domestic species and from animal husbandry practices. It generally does not explore the same scientific questions as research on human biology, even when the same pathogens are being studied, and the discipline is rooted in a very specific agricultural and economic context.Generic and methodological synergies nevertheless exist with biomedical research, particularly with regard to tools and biological models. Certain domestic species furthermore present more functional similarities with humans than laboratory rodents.The singularity of animal health research in relation to biomedical research should be taken into account in the organization, evaluation, and funding of the field through a policy that clearly recognizes the specific issues at stake. At the same time, the One Health approach should facilitate closer collaboration between biomedical and animal health research at the level of research teams and programmes.     

Stratum corneum removal facilitates experimental sensitization to mite allergens in atopic dogs

In humans with atopic dermatitis and in mouse models of IgE-mediated allergic diseases, evidence is mounting that the stratum corneum (SC) provides an important barrier against environmental allergens. At this time, it is not known whether the SC has a similar role in dogs, especially in those with atopic dermatitis. The objectives of this pilot study were to determine whether SC removal led to earlier and stronger sensitization of atopic dogs to Dermatophagoides farinae (Df) house dust mites. Five Maltese-beagle atopic (MBA) dogs were sensitized epicutaneously after the SC was removed with ten tape strips (TS group), while sensitization was done without tape strips in five other MBA dogs (nontape stripping; NTS group). During this 16 week study, sensitization was assessed with allergen-specific IgE serology, intradermal testing with Df allergens and determination of stimulation indices of blood mononuclear cells cultured with Df and stained for CD4 and the activation markers CD25 or CD30. Compared with dogs from the NTS group, those of the TS group exhibited earlier rises in Df-specific IgE serum levels, usually had higher allergen-specific IgE titres, showed higher intradermal test reactivity and had earlier increases and higher percentages of CD25- or CD30-positive activated allergen-specific peripheral CD4-positive T lymphocytes. These observations implicate a role of the SC as a barrier limiting sensitization to exogenous allergens in this experimental atopic dog model.     

Establishment of a quantitative ELISA for the measurement of allergen-specific IgE in dogs using anti-IgE antibody cross-reactive to mouse and dog IgE

As IgE plays a pivotal role in type I hypersensitivity-mediated allergic diseases, it is valuable to measure absolute quantity of serum antigen-specific IgE for clinical and research purposes. Here we describe a novel ELISA system that enables quantification of antigen-specific IgE in ng/ml in dogs. A newly developed monoclonal antibody (CRE-DM) was shown to recognize canine and mouse IgE equally in a dose dependent manner, but it did not recognize canine IgG. The reactivity of CRE-DM to canine IgE was also confirmed by an inhibition ELISA using canine IgE as an inhibitor and the maximum inhibition rate was 91.3%. In order to know whether canine IgE specific to an allergen could be quantitatively measured with an ELISA using CRE-DM, we established a quantitative ELISA that could measure canine IgE recognizing Cry j 1, one of the major allergens of Japanese cedar pollen. In this ELISA, a standard curve was created by using concentration-predetermined Cry j 1-specific monoclonal mouse IgE. According to the standard curve, the concentration of Cry j 1-specific IgE in dogs that were experimentally sensitized to Japanese cedar pollen could be calculated and determined in ng/ml. The specificity of the Cry j 1-specific IgE ELISA using CRE-DM was also confirmed by inhibition ELISA using canine IgE as an inhibitor and the inhibition rate was 97.0%. Reproducibility of the ELISA in three independent assays was determined using groups of pooled canine sera whose Cry j 1-IgE titers ranged from 155.9 to 888.2 ng/ml. Intra- and inter-assay reproducibility was determined with coefficient of variation ranging between 3.1-5.2% and 2.2-8.0%, respectively. These results demonstrated that the ELISA utilizing CRE-DM was a specific, reliable and robust new laboratory test that could quantify absolute amount of antigen-specific IgE in canine serum. The ELISA will serve as a useful tool in the clinics to evaluate the change of serum IgE titers during anti-allergic treatments as well as during seasonal fluctuation of allergen exposure.     

Measurement of systemic and local respiratory cell-mediated immunity after influenza infection in chickens

The detection of ChIFN production after ex vivo antigenic-stimulation of T-lymphocytes has been evaluated for the first time, as a tool to assess cell-mediated immunity (CMI) after avian influenza (AI) infection in 10-day-old SPF chickens. Preliminarily, recall antigens have been produced either by concentrating and inactivating the whole virus or by dissociating the viral proteins. Biologically and structurally intact forms of the viral proteins were isolated by non-ionic detergents while heating, chemical agents and ionic detergent used for virus inactivation altered the antigenic viral components. The n-octyl-B-D-gluco-pyranoside treatment at low temperature was very efficient to produce AI antigenic proteins used for evaluation of ChIFN production after ex vivo antigenic-stimulation of splenic and peripheral lymphocytes. In addition, protocols to isolate lymphocytes from the respiratory tract - the trachea and the lung - have been adapted for local CMI evaluation after similar ex vivo recall assay. Specific AI CMI in the spleen, the blood and the lung was detected for 5 weeks after low pathogenic AI (LPAI) infection in chickens, while further development is needed for tracheal CMI measurement.     

Effects of exogenous amylase on the in vitro digestion kinetics of whole-crop maize silages made from flint or dent grain type at different phenological stages grown in tropical condition

The effect of exogenous amylase on the in vitro rumen digestion kinetics of whole-crop maize silage made from dent (RB9004) or flint grain type (RB9308) was evaluated at different phenological stages: soft dough (SOD), early dent (EAD), ½ milkline (½M) and ¾ milkline (¾M). Forage was harvested from 70 to 110 days after sowing. Two rumen-cannulated cows receiving or not exogenous amylase (0.7 g/kg dry matter—DM, provided to achieve 396 kilo Novo units of amylase activity/kg of TMR DM) were used as donor of ruminal fluid. The in vitro gas production kinetics was evaluated according to a dual-pool logistic model. The chemical composition and gas production kinetics were affected by the hybrid and phenological stages. The flint hybrid had lower range for chemical analysis among physiological stages. Harvesting at ½M and ¾M improved DM content, bromatological composition and silage quality parameters compared to dent or flint types. Amylase (i) increased methane (CH₄) production and in vitro dry matter digestibility (IVDMD) in ½M stage, (ii) improved digestion kinetics by reducing lag time and increasing total gas production and fermentation rates of non-fibrous carbohydrates (NFC) and fibrous carbohydrates (FC), and (iii) increased extent and fermentation rate of NFC and increased fermentation rate of FC fraction in whole-crop maize silages produced from dent or flint types in all phenological stages. Harvesting between ½M and ¾M is the best phenological stage to improve chemical composition and silage quality parameters. Exogenous amylase showed improvements on fibre digestion of silages at ½M and ¾M phenological stages in both grain types of corn.     

Global analysis of Coffea canephora Pierre ex Froehner (Rubiaceae) from the Guineo-Congolese region reveals impacts from climatic refuges and migration effects

As the second species used for commercial coffee, evaluation of Coffea canephora Pierre ex Froehner population genetics is a challenging task for coffee breeding. This study examined the pattern of genetic variability and genetic relationships of cultivated and wild populations of C. canephora sampled across the Guineo-Congolese region of Africa and some improved populations maintained in field genebanks. A total of 293 individuals, sampled from 17 populations, were genotyped with 39 nuclear microsatellite markers. Genetic diversity and structure were investigated with both a model-based and a graphical approach; isolation by distance was also tested. Relationships between the diversity clusters are discussed with regard to differentiation due to several glacial refuges during the Last Glacial Maximum (LGM). High genetic diversity within C. canephora is confirmed with a mean number of alleles of 11.85 per marker, a mean gene diversity of 0.72 and a mean observed heterozygosity of 0.36. An overall structure of two main groups (Guinean and Congolese) subdivided in six subgroups (2 for the Guinean and 4 for the Congolese) was found, including one described for the first time in the Guinean group. A fine structure within the Guinean group was also newly detected. Genetic structure of C. canephora appears to be consistent with its geographic repartition at the continent scale. Structure of diversity was found in accordance with localizations of refuge zones during LGM and migration from this period. Results from this genetic structure study raise our capabilities to better manage and use the collections of genetics resources for breeding purposes. Those results will be used in future association studies to optimize the number of genotypes to be phenotyped.     

Evaluation of sugar beet pulp efficiency for improving the retention of copper in stormwater basin

Purpose

Sugar beet pulp (SBP) has been shown to be a promising low-cost sorbent for the removal of metals from contaminated waters. The objective of this study was to investigate the impact of SBP addition to horizontal-flow gravel filters in increasing the copper (Cu) retention ability of stormwater basins.

Material and methods

Small-scale gravel filters filled with 6 kg of a sand–sediment mix (SS) were used to experimentally treat Cu-contaminated water under conditions that represented an intense storm event. Sugar beet pulp efficiency was assessed by adding 100 g of raw SBP. Two ways of applying SBP were tested: (1) mixed with SS into the gravel filter or (2) packed at the outlet in a PVC column. Eluates were characterized by their volume, pH, Cu and dissolved organic carbon (DOC) concentrations.

Results and discussion

When placed at the outlet, SBP fixed 73 % of the Cu remaining in solution and increased the overall retention capacity of the gravel filter to 99.4 %. Conversely, when SBP was mixed with SS, the outflowing water carried higher concentrations of Cu and DOC. Complementary batch experiments underlined the crucial role of DOC in the decline of Cu sorption ability observed when SS and SBP were mixed. Geochemical calculations suggested that DOC (assumed to be pectins) promotes the mobilisation of Cu from SS by complexing it in porewater.

Conclusions

Accompanied with careful guidance, SBP has the potential of removing dissolved Cu from contaminated water in gravel filters. Protocols for SBP preparation and conditions of use should be established so as to promote its sorption efficiency and decrease its release of Cu-complexing compounds like pectins.     

Zn, Cu, Cd and Hg binding to metallothioneins in harbour porpoises Phocoena phocoena from the southern North Sea

Background  

Harbour porpoises Phocoena phocoena from the southern North Sea are known to display high levels of Zn and Hg in their tissues linked to their nutritional status (emaciation). The question arises regarding a potential role of metallothioneins (MTs) with regard to these high metal levels. In the present study, metallothionein detection and associated Zn, Cd, Cu and Hg concentrations were investigated in the liver and kidney of 14 harbour porpoises collected along the Belgian coast.