Tom Luyten

Interactions between genes involved in growth and muscularity in pigs: IGF-2, myostatin, ryanodine receptor 1, and melanocortin-4 receptor.

In the swine breeding industry, two economical traits are of particular importance in sires, namely, muscle growth and average daily gain (ADG). These traits are quantitative, which implies that they are under the control of multiple genes. Mutations in these genes, associated with either muscularity or growth, are useful quantitative trait nucleotides (QTN) for unraveling genetic variation of these traits and can be used in marker-assisted selection. Until now, QTN involved in muscle growth and/or ADG in pigs were identified in porcine ryanodine receptor 1 (RYR1), insulin-like growth factor-2 (IGF-2), and melanocortin-4 receptor (MC4R). Recently, a fourth possible QTN was found in porcine myostatin (MSTN). All four QTN have an influence on muscle growth and/or somatic growth, so an influence of one mutation on one or more of the other mutations should not be excluded. However, although the polymorphisms in the RYR1 and the MC4R gene affect the function of the respective protein, the polymorphisms of the IGF-2 and MSTN gene influence the mRNA expression of the respective gene. Therefore, this study investigated possible interactions between the genotypes of MSTN, IGF-2, and MC4R (population 1) or the RYR1, IGF-2, and MSTN QTN (population 2) on IGF-2 and MSTN expression in different muscle types in pigs. In both skeletal muscle and heart muscle growth, the IGF-2:MSTN ratio seems to play an important role. Also, the RYR1 genotype had a significant effect on IGF-2 expression in m. longissimus dorsi. No effect of the MC4R QTN could be seen.

Escherichia coli ghosts or live E. coli expressing the ferri-siderophore receptors FepA, FhuE, IroN and IutA do not protect broiler chickens against avian pathogenic E. coli (APEC).

The aim of this study was to investigate if immunization with the ferri-siderophore receptors FepA, FhuE, IroN and IutA could protect chickens against avian pathogenic Escherichia coli (APEC) infection. The antigens were administered as recombinant proteins in the outer membrane (OM) of E. coli strain BL21 Star DE3. In a first immunization experiment, live E. coli expressing all 4 recombinant ferri-siderophore receptors (BL21(L)) were given intranasally. In a second immunization experiment, a mixture of E. coli ghosts containing recombinant FepA and IutA and ghosts containing recombinant FhuE and IroN was evaluated. For both experiments non-recombinant counterparts of the tentative vaccines were administered as placebo. At the time of challenge, the IgG antibody response for BL21(L) and a mixture of E. coli ghosts containing recombinant FepA and IutA and ghosts containing recombinant FhuE and IroN was significantly higher in all immunized groups as compared to the negative control groups (LB or PBS) confirming successful immunization. Although neither of the tentative vaccines could prevent lesions and mortality upon APEC infection, immunization with bacterial ghosts resulted in a decrease in mortality from 50% (PBS) to 31% (non-recombinant ghosts) or 20% (recombinant ghosts) and these differences were not found to be significant.

Bioluminescent avian pathogenic Escherichia coli for monitoring colibacillosis in experimentally infected chickens

Avian pathogenic Escherichia coli (APEC) are responsible for significant economic losses in the poultry industry. In this study, a model for investigating the pathogenesis of APEC infections was established. APEC strain CH2 (O78) was marked with the luciferase operon (luxCDABE) using a Tn7 transposon and tissues of experimentally infected chickens were analysed for a correlation between the bioluminescent signal and the number of bacteria. Transposition of the lux operon into the chromosome of the APEC isolate did not affect sensitivity to lytic bacteriophages and there was no effect on virulence in an intratracheal infection model in 1-day-old chicks, although results with a subcutaneous infection model were inconclusive. A correlation between the number of bacteria and the luminescent signal was found in liquid medium, as well as in homogenised heart, liver, spleen and lung of 4-week-old experimentally infected chickens. This study showed that lux could be used for identification of the infecting strain after experimental infection with APEC in poultry.