Genetics and disease resistance

Genetic components of disease resistance have been described in most of important diseases in human as well as in laboratory and livestock animals. However the basic mechanisms have been established in a few examples only. The reasons herefore are the mostly polygenic inheritance of disease resistance traits, the missing of suitable animal models and the dominance of environmental effects like infection pressure, immune status, and stressors, limiting the view on responsible gene variants. Ethical and practical aspects may further hinder research on disease resistance in certain species. Livestock animals play a crucial role in disease resistance research, because of distinct genetic diversity within and between breeds, because of an often distinct metabolic congruency with humans, and aiming towards the improvement of hygiene and economy of production and animal welfare. The following sections will review disease resistance in livestock animals and their practical implications, completed by examples of our own research activities.     

Water transport in the kidney and nephrogenic diabetes insipidus

Nephrogenic diabetes insipidus is caused by an inability of the kidney to concentrate urine despite adequate concentration of vasopressin in blood and is characterized by polyuria, polydipsia, and hyposthenuria in the presence of plasma hyperosmolality. Nephrogenic diabetes insipidus is the result of defects in water homeostasis in the kidney. Nephrogenic diabetes insipidus occurs when the kidneys cannot or do not respond to vasopressin. There are 2 categories of nephrogenic diabetes insipidus. Congenital nephrogenic diabetes insipidus is a rare, inherited, irreversible cause of polyuria and polydipsia in humans that is even rarer in animals. Acquired nephrogenic diabetes insipidus is more common and is often secondary to illness or medication that interferes with the action of vasopressin in the renal tubules. Unlike congenital nephrogenic diabetes insipidus, acquired or secondary nephrogenic diabetes insipidus is often reversible with correction of the associated or causative problem.     

Analysis of bacterial load and prevalence of mixed infections with Lawsonia intracellularis, Brachyspira hyodysenteriae and/or Brachyspira pilosicoli in German pigs with diarrhoea

Lawsonia (L.) intracellularis, Brachyspira (B.) hyodysenteriae and B. pilosicoli are important pathogens in domestic pig production world-wide, responsible for porcine intestinal adenomatosis, swine dysentery, and porcine intestinal spirochetosis, respectively. Conventional PCR is the major diagnostic tool in the detection of the three pathogens, but the sole detection of bacterial DNA might lead to misinterpretations of results with respect to their clinical relevance, especially with mixed infections. Thus, the present study targeted the detection and quantification of the three pathogens in samples from herds with a case history of diarrhoea. Herds and samples were selected by the practitioners on a voluntary basis. Results were based on 1176 individual samples from 95 herds from Southern Germany. The pathogens were detected simultaneously by multiplex real-time PCR. The overall prevalence for L. intracellularis, B. hyodysenteriae and B. pilosicoli was 12.6%, 8.4% and 3.2% in faecal samples and 48.4%, 24.2% and 31.6% in herds, respectively. Sixty one percent, 82.6%, and 73.4% of herds positive for L. intracellularis, B. hyodysenteriae, and B. pilosicoli, respectively, had mixed infections. Median log values of DNA equivalents/g of faeces for L. intracellularis, B. hyodysenteriae and B. pilosicoli were 3.3, 5.9 and 3.2, with maxima of 8.3, 8.0 and 6.3, respectively. Within herd prevalence of B. hyodysenteriae and B. pilosicoli as well as the load of B. hyodysenteriae were significantly associated with the severity of diarrhoea.     

Ultra‐frequent HRAS p.Q61R somatic mutation in canine acanthomatous ameloblastoma reveals pathogenic similarities with human ameloblastoma

Ameloblastoma is a locally aggressive odontogenic tumour that occurs in humans and dogs. Most ameloblastomas (AM) in humans harbour mutually‐exclusive driving mutations in BRAF, HRAS, KRAS, NRAS or FGFR2 that activate MAPK signalling, and in SMO that activates Hedgehog signalling. The remarkable clinical and histological similarities between canine acanthomatous ameloblastoma (CAA) and AM suggest they may harbour similar driving mutations. In this study, aimed at characterizing the mutational status of SMO, BRAF, HRAS, KRAS, NRAS and FGFR2 in CAA, we used RNA sequencing, Sanger sequencing and restriction fragment length polymorphism assays to demonstrate that 94% of CAA (n = 16) harbour a somatic HRAS p.Q61R mutation. The similarities in MAPK‐activating mutational profiles between CAA and AM implicate conserved molecular mechanisms of tumorigenesis, thus, qualifying the dog as a potentially useful model of disease. Given the relevance of RAS mutations in the pathogenesis of odontogenic tumours and other types of cancer, the results of this study are of comparative, translational, and veterinary value.     

A molecular approach for identification of paramphistomes from Africa and Asia

Paramphistomiasis is a disease of domestic and wild ruminants caused by some digenetic trematodes of the superfamily Paramphistomoidea. Human infections with certain species of paramphistomes have been reported. Given their ubiquity and their abundance within hosts, it seems likely that the importance of these worms is underestimated globally. Based on the reported difficulties of morphological identification of paramphistomes, the development of alternative approaches to better delineate species is needed. Paramphistome samples examined during this study were cercariae, rediae from snail hosts and adult flukes obtained from slaughterhouses from Kenya, Tanzania, Egypt and Nepal. The results confirm that ITS2 is a good molecular marker for paramphistome identification and can be used to determine the affiliation of samples among the different taxa of Paramphistomoidea. Our 22 samples include eight species of paramphistomes. Ten samples of paramphistome larvae (cercariae or rediae) yielded seven species and the 12 samples of adult flukes from bovines yielded four, only one of which was not also represented among the larvae. We found three cases in which the sequences for adult worms and larvae matched, thus revealing the hosts involved locally in those particular life cycles. We interpret our specimens to include five species from Kenya, two from Nepal, two from Tanzania and one from Egypt. Bulinus forskalii was found infected with three species of paramphistomes. The results of the present study will help construct a framework to facilitate identification and eventually control of this underestimated cosmopolitan group of digeneans.     

Interferon-gamma response of PBMC indicates productive pseudorabies virus (PRV) infection in swine

In Chinese Meishan/German Landrace cross-bred swine F2 generation interferon gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) was determined directly ex vivo at different time points after survival of a virulent pseudorabies virus (PRV) infection. This reactivity was compared with the reactivity of na?ve PBMC. Significant IFN-gamma production was determined in ELISA and ELISPOT only after in vitro PBMC re-stimulation with PRV and not with the closely related bovine herpesvirus BHV-1. The PRV-specific IFN-gamma secretion from re-stimulated PBMC showed high levels 6 days after infection, before the presence of serum antibodies, and it persisted at a high level over a 3 months period. The response of a group of eight piglets infected intranasally with PRV varied. Only two animals showed the expected typical fever response. PRV specific IFN-gamma production by PBMC clearly indicated that infection had occurred. Early significant IFN-gamma production by primed PBMC turned out to be a reliable and specific ex vivo marker for cellular response against productive PRV infection in swine before antibody formation.     

A mechanical comparison of 4.5 mm narrow and 3.5 mm broad plating systems for stabilization of gapped fracture models

OBJECTIVE: To evaluate and compare the mechanical properties of 4.5 narrow and 3.5 broad plating systems using their respective cortical and cancellous screws in unstable, central, and eccentric gap fracture models. STUDY DESIGN: Mechanical evaluation and comparison of 2 dynamic compression plate (DCP) systems. SAMPLE POPULATION: Eighteen cortical and 30 cancellous gapped fracture models. METHODS: DCP (4.5 mm narrow, 3.5 mm broad) with their respective cortical screws were applied to cortical bone density polyurethane foam blocks to construct center gap cortical fracture models that were tested in gap closing monotonic 4-point bending. DCP (4.5 mm narrow, 3.5 mm broad) with their respective cancellous screws were applied to cancellous bone density polyurethane foam blocks to construct eccentric gap cancellous fracture models. The cancellous constructs were tested in monotonic gap opening and gap closing cantilever bending and in cyclic axial loading. Univariate and multivariate repeated measures ANOVA were used to compare the maximum loads at failure of the 4.5 mm constructs and 3.5 mm constructs. RESULTS: The 4.5 mm narrow plating system withstood significantly higher loads at failure than the 3.5 mm broad plating system in 4-point bending (P<.0001) and gap opening cantilever bending (P<.0001). The 4.5 mm system failed in gap closing cantilever bending by plastic deformation of the plate, whereas the 3.5 mm system failed by screw pullout. There was no difference between the 2 systems in cyclic axial loading. CONCLUSION: Results indicate that the 4.5 mm narrow plating system has a mechanical advantage over the 3.5 mm broad plating system for stabilization of gapped fracture models. CLINICAL RELEVANCE: The 4.5 mm narrow plating system may be mechanically advantageous compared with the 3.5 mm broad plating system for stabilizing unreconstructed comminuted long bone fractures in large dogs.