A comparison of two methods to assess apparent total tract digestibility of nutrients in dogs

ABSTRACT

The apparent total tract digestibility (ATTD) of nutrients can be assessed by total collection of faeces (TC), which is the reference method, or by the indicator method (IM). Little information is available on proper faecal sampling methodologies for IM in canines to obtain results comparable to TC. The objective of this study was to determine the minimum number of sub-samples required for the IM to make it comparable with TC. A total of 11 adult male dogs were individually housed in metabolism cages. Dogs had access to a grass yard to facilitate defaecation. Faecal sub-samples (1/day) were taken from the daily faecal output to prepare the pooled samples for IM, obtaining cumulative sample combinations of 3 (IM3), 4 (IM4), 5 (IM5), 6 (IM6) and 7 d (IM7). Digestibility of dry matter, gross energy, crude protein and crude fibre was similar between TC and IM5, IM6 and IM7 (p > 0.05). The IM7 presented the greatest statistical similarity with TC. Nevertheless, IM was not a good predictor of crude fibre digestibility. In conclusion, IM can replace the TC method in dogs to evaluate ATTD of several nutritional fractions as long as the composite sample is collected during seven consecutive days. For estimation of fibre digestibility by IM, longer collection periods are probably required.     

Simple recessive mutation in ENAM is associated with amelogenesis imperfecta in Italian Greyhounds

We report a familial enamel hypoplasia in Italian Greyhounds resembling non‐syndromic autosomal recessive amelogenesis imperfecta (AI) of humans. The condition uniformly affects deciduous and permanent teeth and is manifested by enamel roughening/thinning and brownish mottling. Affected teeth are often small and pointed with increased gaps. However, basic tooth structure is usually maintained throughout life, and fractures and dental cavities are not a serious problem as in humans. No tissues or organs other than teeth were affected by this mutation, and there was no relationship between enamel hypoplasia and either autoimmunity or periodontal disease, which also are prevalent in the breed. The enamel hypoplasia was associated with a 5‐bp deletion in exon 10 of the enamelin (ENAM) gene. The prevalence of the enamel defect in Italian Greyhounds was 14%, and 30% of dogs with normal teeth were carriers. Genome analyses suggest that the trait is under inadvertent positive selection. Based on the deletion detected in the ENAM gene, a genetic test was developed for identifying mutation carriers, which would enable breeders to manage the trait.     

A single base deletion in the SLC45A2 gene in a Bullmastiff with oculocutaneous albinism

Oculocutaneous albinism type 4 (OCA4) in humans and similar phenotypes in many animal species are caused by variants in the SLC45A2 gene, encoding a putative sugar transporter. In dog, two independent SLC45A2 variants are known that cause oculocutaneous albinism in Doberman Pinschers and several small dog breeds respectively. For the present study, we investigated a Bullmastiff with oculocutaneous albinism. The affected dog was highly inbred and resulted from the mating of a sire to its own grandmother. We obtained whole genome sequence data from the affected dog and searched specifically for variants in candidate genes known to cause albinism. We detected a single base deletion in exon 6 of the SLC45A2 gene (NM_001037947.1:c.1287delC) that has not been reported thus far. This deletion is predicted to result in an early premature stop codon. It was confirmed by Sanger sequencing and perfectly co‐segregated with the phenotype in the available family members. We genotyped 174 unrelated dogs from diverse breeds, all of which were homozygous wildtype. We therefore suggest that SLC45A2:c.1287delC causes the observed oculocutaneous albinism in the affected Bullmastiff.     

Can Coyotes Affect Deer Populations in Southeastern North America?

ABSTRACT The coyote (Canis latrans) is a recent addition to the fauna of eastern North America, and in many areas coyote populations have been established for only a decade or two. Although coyotes are known predators of white-tailed deer (Odocoileus virginianus) in their historic range, effects this new predator may have on eastern deer populations have received little attention. We speculated that in the southeastern United States, coyotes may be affecting deer recruitment, and we present 5 lines of evidence that suggest this possibility. First, the statewide deer population in South Carolina has declined coincident with the establishment and increase in the coyote population. Second, data sets from the Savannah River Site (SRS) in South Carolina indicate a new mortality source affecting the deer population concurrent with the increase in coyotes. Third, an index of deer recruitment at SRS declined during the period of increase in coyotes. Fourth, food habits data from SRS indicate that fawns are an important food item for coyotes during summer. Finally, recent research from Alabama documented significant coyote predation on fawns there. Although this evidence does not establish cause and effect between coyotes and observed declines in deer recruitment, we argue that additional research should proactively address this topic in the region. We identified several important questions on the nature of the deer—coyote relationship in the East.     

A second KRT71 allele in curly coated dogs

Major characteristics of coat variation in dogs can be explained by variants in only a few genes. Until now, only one missense variant in the KRT71 gene, p.Arg151Trp, has been reported to cause curly hair in dogs. However, this variant does not explain the curly coat in all breeds as the mutant ¹⁵¹Trp allele, for example, is absent in Curly Coated Retrievers. We sequenced the genome of a Curly Coated Retriever at 22× coverage and searched for variants in the KRT71 gene. Only one protein‐changing variant was present in a homozygous state in the Curly Coated Retriever and absent or present in a heterozygous state in 221 control dogs from different dog breeds. This variant, NM_001197029.1:c.1266_1273delinsACA, was an indel variant in exon 7 that caused a frameshift and an altered and probably extended C‐terminus of the KRT71 protein NP_001183958.1:p.(Ser422ArgfsTer?). Using Sanger sequencing, we found that the variant was fixed in a cohort of 125 Curly Coated Retrievers and segregating in five of 14 additionally tested breeds with a curly or wavy coat. KRT71 variants cause curly hair in humans, mice, rats, cats and dogs. Specific KRT71 variants were further shown to cause alopecia. Based on this knowledge from other species and the predicted molecular consequence of the newly identified canine KRT71 variant, it is a compelling candidate causing a second curly hair allele in dogs. It might cause a slightly different coat phenotype than the previously published p.Arg151Trp variant and could potentially be associated with follicular dysplasia in dogs.     

Autoantibodies recognizing proteins copurified with PCNA in patients with connective tissue diseases

Objective. Proliferating cell nuclear antigen (PCNA), one of the target antigen recognized by lupus sera, has been reported to be present as a subnuclear multi-peptide complex. But autoantibodies reacting with components of PCNA complex are poorly understood. To study the specificity of those autoantibodies, immunoreactivities of autoimmune sera against purified PCNA antigen were studied. Methods. PCNA antigens were purified from rabbit thymus extract by affinity column using murine monoclonal antibodies (mAbs) to PCNA, TOB7, TO17 and TO30. Immunoreactivities of autoimmune sera against purified PCNA were analyzed by WB. Results. PCNA antigen purified by serum AK predominantly showed a 34 kD band specific for PCNA in SDS-PAGE. When antigens were purified by anti-PCNA mAb TOB7 and TO30 which are known to be targeting different epitopes on PCNA antigen, SDS-PAGE analysis showed various mol. wt of proteins in addition to the 34 kD PCNA while both AK and mAbs reacted only with 34 kD PCNA in WB. In WB using PCNA purified by TOB7, various immunoreactivities were observed at 150, 66, 58, 48, 45, 37, 32 and 16 kDa in sera from patients with connective tissue diseases. Conclusions. These results suggested that many of the proteins copurified with PCNA were also targets of autoimmune responses and these autoantibody experssion may be induced through antigen-driven mechanisms.Abbreviations mAb monoclonal antibody - PCNA proliferating cell nuclear antigen - PCNA/AK PCNA affinity purified by antibodies from patient serum AK - PCNA/TO30 PCNA purfied by mAb TO30 - PCNA/TOB7 PCNA purified by mAb TOB7 - SLE systemic lupus erythematosus