Non-additive association analysis using proxy phenotypes identifies novel cattle syndromes.

Mammalian species carry ~100 loss-of-function variants per individual1,2, where ~1-5 of these impact essential genes and cause embryonic lethality or severe disease when homozygous3. The functions of the remainder are more difficult to resolve, although the assumption is that these variants impact fitness in less manifest ways. Here we report one of the largest sequence-resolution screens of cattle to date, targeting discovery and validation of non-additive effects in 130,725 animals. We highlight six novel recessive loci with impacts generally exceeding the largest-effect variants identified from additive genome-wide association studies, presenting analogs of human diseases and hitherto-unrecognized disorders. These loci present compelling missense (PLCD4, MTRF1 and DPF2), premature stop (MUS81) and splice-disrupting (GALNT2 and FGD4) mutations, together explaining substantial proportions of inbreeding depression. These results demonstrate that the frequency distribution of deleterious alleles segregating in selected species can afford sufficient power to directly map novel disorders, presenting selection opportunities to minimize the incidence of genetic disease.

Evaluation of infrared thermography as a non-invasive method of measuring the autonomic nervous response in sheep.

Eye temperature measured using infrared thermography (IRT) can be used as a non-invasive measure of autonomic nervous system (ANS) activity in cattle. The objective of this study was to evaluate if changes in eye temperature (measured using IRT) can be used to non-invasively measure ANS activity in sheep. Twenty, 2 to 4-year-old, Romney ewes were randomly assigned to receive either epinephrine (EPI) or physiological saline (SAL) for 5 min administered via jugular catheter (n = 10 ewes/treatment). Eye temperature (°C) was recorded continuously using IRT for approximately 25 min before and 20 min after the start of infusion. Heart rate and heart rate variability, measured using the root mean square of successive differences (RMSSD) and the standard deviation of all inter-beat intervals (SDNN), were recorded for 5 min before and up to 10 min after the start of infusion. Blood samples were taken before and after the infusion period to measure plasma epinephrine, norepinephrine, cortisol and packed cell volume (PCV) concentrations. During the infusion period, maximum eye temperature was on average higher (P<0.05) in sheep that received epinephrine than those that received saline. On average, heart rate was higher (SAL: 87.5 beats/min, EPI: 123.2 beats/min, SED = 7.07 beats/min; P<0.05), and RMSSD (SAL: 55.3 ms, EPI: 17.3 ms, SED = 14.18 ms) and SDNN (SAL: 54.3 ms, EPI: 21.5 ms, SED = 10.00 ms) lower (P<0.05) in ewes during the 5 min post-infusion period compared with ewes that received saline. An infusion of epinephrine resulted in higher geometric mean epinephrine (P<0.05) and cortisol (P<0.05) but not norepinephrine (P>0.05) concentrations in ewes compared to an infusion of saline. PCV concentrations were higher (P<0.001) by 7 ± 1.0% (mean±SED) in ewes after an epinephrine infusion. These results suggest that heart rate variability is a sensitive, non-invasive method that can be used to measure ANS activity in sheep, whereas change in eye temperature measured using IRT is a less sensitive method.

Genome-wide association analysis reveals QTL and candidate mutations involved in white spotting in cattle.

BACKGROUND: White spotting of the coat is a characteristic trait of various domestic species including cattle and other mammals. It is a hallmark of Holstein-Friesian cattle, and several previous studies have detected genetic loci with major effects for white spotting in animals with Holstein-Friesian ancestry. Here, our aim was to better understand the underlying genetic and molecular mechanisms of white spotting, by conducting the largest mapping study for this trait in cattle, to date. RESULTS: Using imputed whole-genome sequence data, we conducted a genome-wide association analysis in 2973 mixed-breed cows and bulls. Highly significant quantitative trait loci (QTL) were found on chromosomes 6 and 22, highlighting the well-established coat color genes KIT and MITF as likely responsible for these effects. These results are in broad agreement with previous studies, although we also report a third significant QTL on chromosome 2 that appears to be novel. This signal maps immediately adjacent to the PAX3 gene, which encodes a known transcription factor that controls MITF expression and is the causal locus for white spotting in horses. More detailed examination of these loci revealed a candidate causal mutation in PAX3 (p.Thr424Met), and another candidate mutation (rs209784468) within a conserved element in intron 2 of MITF transcripts expressed in the skin. These analyses also revealed a mechanistic ambiguity at the chromosome 6 locus, where highly dispersed association signals suggested multiple or multiallelic QTL involving KIT and/or other genes in this region. CONCLUSIONS: Our findings extend those of previous studies that reported KIT as a likely causal gene for white spotting, and report novel associations between candidate causal mutations in both the MITF and PAX3 genes. The sizes of the effects of these QTL are substantial, and could be used to select animals with darker, or conversely whiter, coats depending on the desired characteristics.

Effect of Morphine Administration on Social andNon-Social Play Behaviour in Calves.

The objective of this study was to evaluate the effect of morphine on social and non-socialplay behaviour in calves. Twelve calves experienced four treatments in a cross over 2 × 2 factorialdesign: Calves received an intravenous injection of morphine or saline 10 min prior to being testedindividually or in pairs in an arena for 20 min. Play behaviour was continuously recorded in thearena test. Lying times were recorded in the home pen. Cortisol concentrations were measuredbefore and after testing. In the arena test, calves given morphine tended to perform more social playevents than calves given saline, however, morphine administration had no effect on locomotor play.Calves given morphine spent less time lying than calves given saline during the first 4 h afterreturning to the home pen. Cortisol concentrations were suppressed in calves given morphine.Administration of morphine appeared to increase social play but had no effect on locomotor playin calves. This study highlights the importance of investigating different aspects of play behaviourin animals as some may be more indicative of a positive affective state than others. More studiesinvestigating the effects of morphine on play are needed to confirm the results found in this study.