KRT5 in-frame deletion in a family of German Shepherd dogs with split paw pad disease resembling localized epidermolysis bullosa simplex in human patients.

Split paw pad disease is a scarcely defined phenotype characterized by skin lesions on the paw pads of dogs. We studied a family of German Shepherd dogs, in which four dogs developed intermittent paw pad lesions and lameness. The paw pads of two of the affected dogs were biopsied and demonstrated cleft formation in the stratum spinosum and stratum corneum, the outermost layers of the epidermis. Whole genome sequencing data from an affected dog revealed a private heterozygous 18 bp in frame deletion in the KRT5 gene. The deletion NM_001346035.1:c.988_1005del or NP_001332964.1:p.(Asn330_Asp335del) is predicted to lead to a loss of six amino acids in the L12 linker domain of the encoded keratin 5. KRT5 variants in human patients lead to various subtypes of epidermolysis bullosa simplex (EBS). Localized EBS is the mildest of the KRT5-related human diseases and may be caused by variants affecting the L12 linker domain of keratin 5. We therefore think that the detected KRT5 deletion in dogs represents a candidate causal variant for the observed skin lesions in dogs. However, while the clinical phenotype of KRT5-mutant dogs of this study closely resembles human patients with localized EBS, there are differences in the histopathology. EBS is defined by cleft formation within the basal layer of the epidermis while the cleft formation in the dogs described herein occurred in the outermost layers, a hallmark of split paw pad disease. Our study provides a basis for further studies into the exact relation of split paw pad disease and EBS.

The intraruminal redox potential is stabilised by opposing influences during fermentation.

An optimal fermentation process in the forestomach is pivotal for the wellbeing and performance of ruminants. Complex carbohydrates are broken down into short-chain fatty acids (SCFA) which form the major energy source for the animal. A strong interrelationship of this process with intraruminal pH and redox potential (Eh) exists. These parameters can be measured with intraruminal sensors, but the interpretation of the measurements, especially of Eh, and their meaning for intraruminal homeostasis is not completely clear. In this study, factors influencing intraruminal Eh were elucidated. We hypothesised that intraruminal Eh is influenced by the fermentation process as such, but not by its end products SCFA. We measured Eh and pH in ruminal fluid from fasting cannulated sheep after the addition of 0.06 m Na-acetate, -propionate, -butyrate or glucose in vitro. Furthermore, we assessed the interrelation of pH and Eh. Basal Eh and pH values were -120 ± 41 mV and 7.0 ± 0.3, respectively, in native ruminal fluid in vitro. While the addition of SCFA did not induce any changes, glucose addition caused a significant decrease in both pH and Eh compared to the values before the addition (paired Student's t-test, p < 0.05). We attribute the decrease in Eh to an increased production of H2 in the process of generating SCFA, predominantly acetate. By titrating both native and particle-free ruminal fluid to more acidic and basic pH values (4.5-8.5), we found a non-linear inverse correlation of pH and Eh, counteracting the H2 -driven decrease of Eh during fermentation. Thus, the intraruminal Eh is influenced by pH and H2 output during SCFA formation. The opposed character of these factors stabilises the intraruminal homeostasis which might help maintain symbiotic microbiota in the rumen. Understanding, monitoring, and supporting this system will be an essential part of modern cattle production.

EDA Missense Variant in a Cat with X-Linked Hypohidrotic Ectodermal Dysplasia.

Hypohidrotic ectodermal dysplasia is a developmental defect characterized by sparse or absent hair, missing or malformed teeth and defects in eccrine glands. Loss-of-function variants in the X-chromosomal EDA gene have been reported to cause hypohidrotic ectodermal dysplasia in humans, mice, dogs and cattle. We investigated a male cat exhibiting diffuse truncal alopecia with a completely absent undercoat. The cat lacked several teeth, and the remaining teeth had an abnormal conical shape. Whole-genome sequencing revealed a hemizygous missense variant in the EDA gene, XM_011291781.3:c.1042G>A or XP_011290083.1:p.(Ala348Thr). The predicted amino acid exchange is located in the C-terminal TNF signaling domain of the encoded ectodysplasin. The corresponding missense variant in the human EDA gene, p.Ala349Thr, has been reported as a recurring pathogenic variant in several human patients with X-linked hypohidrotic ectodermal dysplasia. The identified feline variant therefore represents the likely cause of the hypohidrotic ectodermal dysplasia in the investigated cat, and the genetic investigation confirmed the suspected clinical diagnosis. This is the first report of an EDA-related hypohidrotic ectodermal dysplasia in cats.