Two MC1R loss-of-function alleles in cream-coloured Australian Cattle Dogs and white Huskies.

Loss-of-function variants in the MC1R gene cause recessive red or yellow coat-colour phenotypes in many species. The canine MC1R:c.916C>T (p.Arg306Ter) variant is widespread and found in a homozygous state in many uniformly yellow- or red-coloured dogs. We investigated cream-coloured Australian Cattle Dogs whose coat colour could not be explained by this variant. A genome-wide association study with 10 cream and 123 red Australian Cattle Dogs confirmed that the cream locus indeed maps to MC1R. Whole-genome sequencing of cream dogs revealed a single nucleotide variant within the MITF binding site of the canine MC1R promoter. We propose to designate the mutant alleles at MC1R:c.916C>T as e1 and at the new promoter variant as e2 . Both alleles segregate in the Australian Cattle Dog breed. When we considered both alleles in combination, we observed perfect association between the MC1R genotypes and the cream coat colour phenotype in a cohort of 10 cases and 324 control dogs. Analysis of the MC1R transcript levels in an e1 /e2 compound heterozygous dog confirmed that the transcript levels of the e2 allele were markedly reduced with respect to the e1 allele. We further report another MC1R loss-of-function allele in Alaskan and Siberian Huskies caused by a 2-bp deletion in the coding sequence, MC1R:c.816_817delCT. We propose to term this allele e3 . Huskies that carry two copies of MC1R loss-of-function alleles have a white coat colour.

Cold thermopeaking-induced drift of nase Chondrostoma nasus larvae.

Research on how intermittent water releases from hydropower plants affect the early life stages of fish has advanced in the last years, focusing not only on the direct impacts of rapid flow changes (hydropeaking), but also on the short-term fluctuations in water temperature (thermopeaking). Flow and thermal fluctuations caused by hydropeaking may affect fish movement patterns and migration at critical stages of a species' life cycle, e.g., by inducing passive downstream drift. Using two experimental outdoor channels, we investigated how nase (Chondrostoma nasus, Cypriniformes) larvae respond to a rapid drop in water temperature during hydropeaking (simulating a cold thermopeaking event), reaching on average 5.5 °C under peak flow (maximum discharge) conditions, in comparison with a hydropeaking treatment with a constant water temperature regime. Responses of fish larvae were analyzed during acclimation, up-ramping (increase in discharge), peak flow and down-ramping (decrease in discharge) phases. Fish drift increased during peak flow in the cold thermopeaking treatment compared to hydropeaking. Higher drift rates were also negatively associated with pronounced water temperature drops during peak flow conditions. In addition, the starting temperature of the experiment influenced drift during up-ramping. Overall, the results suggest that cold thermopeaking may increase drift in the early life stages of cypriniform fish compared with hydropeaking with stable water temperature. Hence, monitoring and active water temperature adjustments following hydropower releases should be adopted as strategies to mitigate power plant-related impacts on aquatic organisms. Supplementary Information: The online version contains supplementary material available at 10.1007/s00027-023-00955-x.

The variant red coat colour phenotype of Holstein cattle maps to BTA27.

The variant red phenotype in Holstein cattle is indistinguishable from the traditional e/e recessive red phenotype caused by a mutation in melanocortin 1 receptor, but is inherited as a dominant trait in relation to black. Co-segregation analysis in four half-sib families segregating for variant red was conducted, excluding melanocortin 1 receptor, agouti signalling protein, attractin and melatonin receptor 1A as causative genes. However, variant red co-segregated with markers in a region of BTA27 that includes beta-defensin 103 (DEFB103). Two newly identified microsatellites and 5 SNPs 5' of DEFB103 were used for linkage mapping in four segregating families (LOD = 3.26). One haplotype was inherited in VR cattle in a 6-generation pedigree.

Genetic variation in the pro-melanin-concentrating hormone gene affects carcass traits in Bos taurus cattle.

Mammalian pro-melanin-concentrating hormone (PMCH) has previously been shown to affect feed intake in rodent species. The objectives of this study were to sequence the Bos taurus PMCH gene in order to identify any existing genetic variants and to evaluate whether these affected carcass traits. An A-to-T SNP was identified at position -134 relative to the ATG start codon (g.-134A>T). The alleles at this SNP were significantly associated with average fat and grade fat in two crossbred populations of Bos taurus cattle. The g.-134T allele may introduce a binding site for the transcriptional repressor, adenovirus E4 promoter binding protein, which may contribute to this effect. The g.-134A allele occurred in 67% of cattle examined and was associated with higher fat levels.

A model-based fish bioassessment index for Eastern Mediterranean rivers: Application in a biogeographically diverse area.

In ecosystems with high fish species endemicity, such as Mediterranean-type rivers, biogeographical differences among ecoregions present serious obstacles to developing broadly-applicable river bioassessment indices. This impediment has contributed to a serious time-lag in developing EU policy-relevant fish-based indices in the Eastern Mediterranean countries. Here we present the first model-based fish index for the Eastern Mediterranean (the Hellenic Fish Index, HeFI) in an effort to overcome biogeographic differences among the area's biotically heterogeneous rivers. The index is based on modelled reference conditions and employs site-specific electrofished fish samples from an extensive dataset from Greece that covers six freshwater ecoregions, including five transboundary river basins flowing through six countries. Environmental and anthropogenic pressure data were procured from 403 sampled river sites and ecologically-relevant traits were defined for 103 collected fish species. For the development of the index, we first diagnosed least degraded sites forming a calibrated reference site dataset and secondly quantified differences of fish metrics between the reference and impaired sites. Four trait-based fish metrics showed the best ability to discriminate between impaired and reference sites. The index performed well in discriminating anthropogenic pressure classes, giving a significant negative linear response to a gradient of anthropogenic degradation. HeFI successfully assessed both small and large rivers in different freshwater ecoregions. This geographically broad-scale index development shows that key trait-based reference conditions can be produced by a predictive model in remarkably heterogeneous rivers where range-restricted fishes dominate. This index promotes a screening-level bioassessment application that may be further developed and refined with relevant monitoring.

Genes affecting coat colour and pattern in domestic dogs: a review.

Tremendous progress has been made in identifying genes involved in pigmentation in dogs in the past few years. Comparative genomics has both aided and benefited from these findings. Seven genes that cause specific coat colours and/or patterns in dogs have been identified: melanocortin 1 receptor, tyrosinase related protein 1, agouti signal peptide, melanophilin, SILV (formerly PMEL17), microphthalmia-associated transcription factor and beta-defensin 103. Although not all alleles have been yet identified at each locus, DNA tests are available for many. The identification of these alleles has provided information on interactions in this complex set of genes involved in both pigmentation and neurological development. The review also discusses pleiotropic effects of some coat colour genes as they relate to disease. The alleles found in various breeds have shed light on some potential breed development histories and phylogenetic relationships. The information is of value to dog breeders who have selected for and against specific colours since breed standards and dog showing began in the late 1800s. Because coat colour is such a visible trait, this information will also be a valuable teaching resource.

Assessing the impact of a downscaled climate change simulation on the fish fauna in an Inner-Alpine River.

This study assesses the impact of a changing climate on fish fauna by comparing the past mean state of fish assemblage to a possible future mean state. It is based on (1) local scale observations along an Inner-Alpine river called Mur, (2) an IPCC emission scenario (IS92a), implemented by atmosphere-ocean global circulation model (AOGCM) ECHAM4/OPYC3, and (3) a model-chain that links climate research to hydrobiology. The Mur River is still in a near-natural condition and water temperature in summer is the most important aquatic ecological constraint for fish distribution. The methodological strategy is (1) to use downscaled air temperature and precipitation scenarios for the first half of the twenty-first century, (2) to establish a model that simulates water temperature by means of air temperature and flow rate in order to generate water temperature scenarios, and (3) to evaluate the impact on fish communities using an ecological model that is driven by water temperature. This methodology links the response of fish fauna to an IPCC emission scenario and is to our knowledge an unprecedented approach. The downscaled IS92a scenarios show increased mean air temperatures during the whole year and increased precipitation totals during summer, but reduced totals for the rest of the annual cycle. These changes result in scenarios of increased water temperatures, an altered annual cycle of flow rate, and, in turn, a 70 m displacement in elevation of fish communities towards the river's head. This would enhance stress on species that rely on low water temperatures and coerce cyprinid species into advancing against retreating salmonids. Hyporhithral river sectors would turn into epipotamal sectors. Grayling (Thymallus thymallus) and Danube salmon (Hucho hucho), presently characteristic for the Mur River, would be superceded by other species. Native brown trout (Salmo trutta), already now under pressure of competition, may be at risk of losing its habitat in favour of invaders like the exotic rainbow trout (Oncorhynchus mykiss), which are better adapted to higher water temperatures. Projected changes in fish communities suggest an adverse influence on salmonid sport fishing and a loss in its high economic value.

Chromosomal aneuploidy associated with spontaneous abortions and neonatal losses in cattle.

Pericardial sac samples from 77 bovine aborted fetuses and stillborn calves were submitted for tissue culture; cells from 55 of these samples were grown successfully in culture. Six of the 55 karyotyped fetuses (10.7%) had an abnormal chromosome complement, in 3 of which (5.5%) the abnormality was probably the cause of death. This level of abnormality is relatively high when one considers that most fetuses were >8 months gestational age. Approximately 5-7% of human stillbirths and 50% of first-trimester aborted fetuses have chromosome anomalies. If a similar situation exists in cattle, as suggested by these data, chromosome abnormalities may be a major cause of early fetal loss in cattle. Most chromosomally abnormal fetuses had multiple malformations, which suggests that the diagnostic use of chromosome analysis is most cost effective for malformed fetuses and newborns. Twins were present in a higher proportion of these fetuses than expected based on their incidence among liveborn cattle.

Comparative efficiency and accuracy of surgical and cytogenetic sexing in psittacines.

Otoscopic surgical sexing and chromosome analysis through lymphocyte culture were undertaken in 22 psittacine birds of eight species for the purpose of establishing their sex. Comparison of the techniques involved considering success rate, quality of determination, cost, efficiency, and risk. Surgical endoscopy appears to be preferable to chromosome analysis in all categories except risk.

Restriction fragment length polymorphism in the mitochondrial DNA of cloned cattle.

The clonal origin of 4 Holstein bulls was determined by hybridization experiments with 2 different minisatellite probes, and all 4 animals showed identical genomic DNA fingerprints, hence confirming monozygosity. Extra-chromosomal differences were observed among these 4 Holstein bulls. Mitochondrial DNA restriction fragment length polymorphisms with restriction endonucleases Avall and Hhal sites were found, and these polymorphisms can be explained by the loss of a single site for each of these 2 enzymes. Since mitochondrial DNA are maternally transmitted, all 4 bulls would produce genetically equivalent spermatozoa and offspring. The combination of embryo cloning and specific cytoplasmic markers would provide an ideal system for the study of maternal cytoplasmic effects on quantitative traits.

Fertility effects of the 14;20 Robertsonian translocation in cattle.

Superovulation and embryo collection procedures were used to study the effect of the 14;20 Robertsonian translocation on fertility and embryo viability. Karyotypes were successfully completed on cells from 77 of the 279 embryos prepared for such analysis. Embryos from 4 cows heterozygous for the translocation were studied. Two bulls with the same condition were studied by using their semen in artificial insemination of cows with normal karyotypes. The proportions of fertilized ova and transferable embryos were not different between cows with the 14;20 translocation and those with normal karyotypes, indicating that fertilization rates were not affected by the translocation. Twenty-two percent of the embryos which were karyotyped had an unbalanced karyotype and would theoretically not have survived to term. All of the theoretically predicted chromosome complements from such a translocation were observed as were three 58,XX,t karyotypes and a 58,XX karyotype. There was no difference in the percentage of embryos with abnormal karyotypes whether the cow or bull was the carrier. Results therefore indicate that fertility is rather severely impaired in carriers of the 14;20 translocation, as was observed with the 1;29 translocation, with most loss due to embryo mortality rather than a lowered conception rate.

Embryonic loss in superovulated cattle caused by the 1;29 Robertsonian translocation.

The effect of the 1;29 Robertsonian translocation on fertility was studied using embryos resulting from matings of nine carrier cows and two carrier bulls. Embryos were collected from the following three mating groups utilizing superovulation: normal bull cross normal cow, normal bull cross translocation carrier cow, and translocation carrier bull cross normal cow. The proportion of ova which were fertilized did not vary among the groups, indicating that fertilization rates were not affected by the translocation. The translocation cows did yield fewer embryos on average than did cows with normal karyotypes, which may suggest ovulation rates are reduced (at least after superovulation attempts) in cattle carrying the 1;29 translocation. Twenty of 39 embryos successfully karyotyped had abnormal chromosome complements. All four of the theoretically predicted karyotypes and two additional abnormal combinations were found. Eight of 39 (20.5%) embryos karyotyped had unbalanced karyotypes which would have resulted in embryonic loss. The proportion of embryos with unbalanced karyotypes, was slightly higher when the cow (36%) carried the translocation than when the bull (19%) did. Results of this study indicate that fertility is impaired due to the presence of this translocation. The major loss in reproductive potential appears to be due to embryonic loss rather than fertilization failure.

Development of the Canadian beef reference herd for gene mapping studies.

A project to map quantitative trait loci (QTL), in beef cattle using a full-sib design was initiated using six Bos taurus breeds. Embryo transfer was used in a large scale, short timeframe experiment to develop this herd for gene mapping. Full-sib families allowed for genetic information to be followed through both the sire and the dam and for both parents to be slaughtered so that carcass quality data could also be obtained from both of them at close to typical slaughter ages. Repeatability of response to superovulation was significant among the 3 flushes per female. Response to superovulation was negatively correlated with backfat of the donor. Crossbred embryos were found to have higher survival than purebred embryos.

A survey of malformed aborted bovine fetuses, stillbirths and nonviable neonates for abnormal karyotypes.

Postmortem examinations were performed on 30 morphologically abnormal aborted bovine fetuses, stillbirths and nonviable neonates. Fibroblasts from the pericardium were cultured for chromosome analysis. Karyotypes were successfully completed on 18 animals, of which three were trisomic, one was mosaic monosomic and one was chimeric. All aneuploid calves had multisystemic anomalies. Using chromosomal banding techniques, the abnormal karyotypes were determined to be: 61,XY,+27; 61,XX,+21; 61,XY,+?; 59,XY,-?/60,XY; and 60,XX/60,XY. Bacterial contamination or nonviability of tissues prevented the growth of fibroblasts in culture and cytogenetic analysis of the other 12 animals. It was estimated that 2.0% of all late gestation abortuses and stillbirths may have chromosomal abnormalities characterized by aneuploidy. The findings of this study suggest chromosomal abnormalities characterized by aneuploidy are a significant cause of multisystemic anomalies in aborted bovine fetuses and nonviable neonates.

Multifocal retinopathy of Great Pyrenees dogs.

Forty-four related Great Pyrenees dogs were examined ophthalmoscopically. Focal retinal elevations, multiple gray-tan-pink subretinal patches, and discrete areas of tapetal hyper-reflectivity were seen in 19 dogs, ranging from 13 weeks to 10 years of age. These lesions varied in size from focal spots that were barely visible with the indirect ophthalmoscope to areas that were larger than the optic disc. Complete blood cell counts, serum biochemical profiles, urinalyses, and blood pressure measurements were completed on four affected dogs and all were within normal reference ranges. Photopic and scotopic electroretinography was completed and the a-wave and b-wave amplitudes and latencies were similar for affected and age-matched nonaffected Great Pyrenees and other normal dogs. Electroretinograms that were examined twice during a 3-year period on three affected adult dogs did not reveal significant progressive deterioration of the a or b-wave parameters. Fluorescein angiography was completed on four affected dogs of ages 1 (n = 2), 5, and 6 years. These angiograms were repeated in three of these dogs 1 year later. The blood ocular barrier was intact in these dogs but there was blocked choroidal fluorescence. Postmortem examination, light microscopy, scanning and transmission electron microscopy were performed on three affected puppies and two affected adult dogs. These examinations revealed that the lesions in the puppies were limited to bilateral multiple areas of retinal pigment epithelial vacuolation, hypertrophy, and apparent separation from Bruch's membrane, and multiple serous retinal detachments. The affected adult dogs had focal retinal degeneration and retinal pigment epithelial hypertrophy, hyperplasia and pigmentation. Pedigree analysis and test mating confirm that this condition is inherited, probably as an autosomal recessive trait. This condition develops at approximately 13 weeks of age and the focal areas of retinal detachment and retinal pigment epithelial vacuolation progress to permanent and stable focal areas of retinal degeneration, and retinal pigment epithelial hypertrophy and pigmentation.

Variability in vocal and behavioural responses to visual isolation between full-sibling families of beef calves.

In order to assess the value of vocal response as an indicator of welfare in cattle, it is necessary to investigate factors which influence vocal behaviour of individuals, that are independent of the specific environmental conditions to which they are exposed. The effect of parental differences, sex, age and weight on vocal responses of calves to visual isolation were examined. The relationship between vocal response and the amount of body movement during isolation were also evaluated.Seventeen full-sibling families of beef calves (N=130) were created by breeding five sires with 13 superovulated dams. Embryos were transferred into unrelated cows, which reared the calves to weaning age. Vocal response of calves and body movement were measured on four occasions, while they were visually isolated for 1min on a single animal scale platform. The first two observations were made on the day of weaning (mean age 166 days) and the following day. At the third and fourth observations their average ages were 278 and 350 days, respectively. On the four occasions 33.3, 34.8, 27.8 and 38.6% of calves, respectively, vocalized. Both sire and family had a significant influence on the number of vocalizations and acoustic properties of the sounds. Electronically-measured movement while on the scale was also influenced by sire and family, but there was no relationship between vocal responses and body movement. There was a tendency (P=0.08) for older and therefore heavier calves to produce longer vocalizations. In the final observing session, age and weight were positively correlated with fundamental frequency and the sound pressure level at the frequency of greatest intensity, and negatively correlated with the loudest harmonic. At the last two observations, significantly more heifer calves than bull calves vocalized. There was little difference between the acoustic properties of heifers' and bulls' vocalizations.The considerable individual variability in vocal response within a group of identically-treated calves is partly due to parental genetic influences that are independent of rearing and early experience. Other endogenous characteristics such as sex, age and weight also influence vocal response. But there is no clear relationship between individual temperament (as assessed by the amount of body movement during isolation) and characteristics of vocal response.

Chromosomal trisomy in an anormalous bovine fetus.

This case report describes an anomalous male fetus of approximately six months gestation with abnormal karyotype containing an extra metacentric chromosome of medium size. The fetus had severe phenotypic abnormalities including anasarca, eye malformations, cardiac hypertrophy, pulmonary hypoplasia, adrenal dysplasia, and an umbilical hernia.

A study of the inheritance of a bleeding disorder in Simmental cattle.

A study was designed to determine the inheritance pattern of a blood platelet aggregation disorder in Simmental cattle utilizing embryo transfer technology. A Simmental donor cow that had previously produced a calf with the platelet aggregation disorder was superovulated and mated to a bull that had also produced affected offspring. Twenty-seven calves were produced from the 63 (42.9%) embryos transferred. This somewhat lower than expected pregnancy rate is suggestive of an increased rate of embryo loss. Twenty-three of 25 (92%) calves had normal platelet aggregation patterns and 2 failed to show any evidence of platelet aggregation. Data are suggestive that inheritance is not simple Mendelian recessive. A more likely scenario is that the defect is the result of the inheritance of at least 2 genes, which is also consistent with the sporadic incidence reported in the population at large.

Black hair follicular dysplasia, an autosomal recessive condition in dogs.

Using histology, a coat color abnormality and the subsequent hair loss were diagnosed as black hair follicular dysplasia. A pedigree analysis of an affected litter and literature review suggests that this is inherited as an autosomal recessive trait. The melanocyte stimulating hormone receptor gene is ruled out by using linkage analysis.