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1.
Anim Genet ; 50(3): 307-310, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30957265

RESUMO

Gender assignment errors are common in some animal species and lead to inaccuracies in downstream analyses. Procedures for detecting gender misassignment are available for array-based SNP data but are still being developed for genotyping-by-sequencing (GBS) data. In this study, we describe a method for using GBS data to predict gender using X and Y chromosomal SNPs. From a set of 1286 X chromosomal and 23 Y chromosomal deer (Cervus sp.) SNPs discovered from GBS sequence reads, a prediction model was built using a training dataset of 422 Red deer and validated using a test dataset of 868 Red deer and Wapiti deer. Prediction was based on the proportion of heterozygous genotypes on the X chromosome and the proportion of non-missing genotypes on the Y chromosome observed in each individual. The concordance between recorded gender and predicted gender was 98.6% in the training dataset and 99.3% in the test dataset. The model identified five individuals across both datasets with incorrect recorded gender and was unable to predict gender for another five individuals. Overall, our method predicted gender with a high degree of accuracy and could be used for quality control in gender assignment datasets or for assigning gender when unrecorded, provided a suitable reference genome is available.


Assuntos
Cervos/genética , Análise para Determinação do Sexo , Animais , Cervos/fisiologia , Feminino , Masculino , Polimorfismo de Nucleotídeo Único , Cromossomo X , Cromossomo Y
2.
Anim Genet ; 49(5): 361-370, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30062723

RESUMO

Wool is composed primarily of proteins belonging to the keratin family. These include the keratins and keratin-associated proteins (KAPs) that are responsible for the structural and mechanical properties of wool fibre. Although all human keratin and KAP genes have been annotated, many of their ovine counterparts remain unknown and even less is known about their genomic organisation. The aim of this study was to use a combinatory approach including comprehensive cDNA and de novo genomic sequencing to identify ovine keratin and KAP genes and their genomic organisation and to validate the keratins and KAPs involved in wool production using ovine expressed sequence tag (EST) libraries and proteomics. The number of genes and their genomic organisation are generally conserved between sheep, cattle and human, despite some unique features in the sheep. Validation by protein mass spectrometry identified multiple keratins (types I and II), epithelial keratins and KAPs. However, 15 EST-derived genes, including one type II keratin and 14 KAPs, were identified in the sheep genome that were not present in the NCBI gene set, providing a significant increase in the number of keratin genes mapped on the sheep genome.


Assuntos
Queratinas/genética , Carneiro Doméstico/genética , Lã/química , Animais , Bovinos , Cromossomos Artificiais Bacterianos , DNA Complementar/genética , Genoma , Folículo Piloso/química , Folículo Piloso/crescimento & desenvolvimento , Humanos , Queratinas/química
3.
Anaesthesia ; 71(1): 76-84, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26566603

RESUMO

Our bi-specialty clinic was established to systematically investigate patients with suspected peri-operative hypersensitivity reactions. Four hundred and ten patients were studied; 316 following an intra-operative reaction ('postoperative' group) and 94 with a previous history of reaction, referred before undergoing anaesthesia ('pre-operative' group). In the postoperative group, 173 (54.7%) were diagnosed with IgE-mediated reactions: 65 (37.6%) to neuromuscular blocking drugs; 54 (31.2%) antibiotics; 15 (8.7%) chlorhexidine and 12 (6.9%) patent blue dye. Reactions were severe in 114 patients (65.9%). All reactions to patent blue dye were severe. We identified IgE sensitisation in 22 (13.2%) cases with isolated mucocutaneous reactions. Only 173 (54.7%) patients had serum tryptase samples taken. Referrers' suspected causal agent was confirmed in only 37.2% of patients. Of 94 patients reviewed 'pre-operatively', 29 (30.8%) were diagnosed with IgE-mediated hypersensitivity reactions, reinforcing the importance of investigating this group of patients. Knowledge of the range of causative agents identified in our study should guide the investigation of suspected peri-operative hypersensitivity reactions.


Assuntos
Anestesia/efeitos adversos , Hipersensibilidade a Drogas/etiologia , Adulto , Feminino , Humanos , Imunoglobulina E/imunologia , Masculino , Pessoa de Meia-Idade , Triptases/sangue
4.
Anim Genet ; 46(2): 185-9, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25662407

RESUMO

The importance of maintaining DNA methylation patterns and faithful transmission of these patterns during cell division to ensure appropriate gene expression has been known for many decades now. It has largely been assumed that the symmetrical nature of CpG motifs, the most common site for DNA methylation in mammals, together with the presence of maintenance methylases able to methylate newly synthesised DNA, ensures that there is concordance of methylation on both strands. However, although this assumption is compelling in theory, little experimental evidence exists that either supports or refutes this assumption. Here, we have undertaken a genome-wide single-nucleotide resolution analysis to determine the frequency with which hemimethylated CpG sites exist in sheep muscle tissue. Analysis of multiple independent samples provides strong evidence that stably maintained hemimethylation is a very rare occurrence, at least in this tissue. Given the rarity of stably maintained hemimethylation, next-generation sequencing data from both DNA strands may be carefully combined to increase the accuracy with which DNA methylation can be measured at single-nucleotide resolution.


Assuntos
Metilação de DNA , Músculo Esquelético/metabolismo , Carneiro Doméstico/genética , Animais , Ilhas de CpG , Biblioteca Gênica
5.
Anim Genet ; 45(4): 559-64, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24804852

RESUMO

Pithomycotoxicosis, more commonly known as facial eczema (FE), is a liver disease that occurs predominantly in New Zealand because of its toxigenic Pithomyces chartarum strains. The first reported case was in sheep in 1887. Since the 1930s, a number of studies have been conducted in an attempt to mitigate the problems FE has on the sheep and dairy industries. The research in these studies included work on fungicide and biological control of the saprophytic fungus, use of different pasture plants to inhibit fungal growth, stock management with respect to pasture fungal spore counts and the use of zinc prophylaxis on animals. The finding that there was a genetic basis in FE sensitivity in sheep prompted research for a genetic approach to mitigation in the form of a diagnostic DNA test for susceptibility to the disease. Recently, we have used the Illumina OvineSNP50 BeadChip to develop a genome-enabled prediction approach to screen for FE-tolerant sheep. Our current best genomic prediction for FE is for the Romney breed and has an accuracy of 0.38. This prediction accuracy is not as high as the individual accuracy gained by an artificial challenge test (0.64). However, it has the advantage of being a non-invasive test and can be provided as part of genomic testing for other traits at minimal cost.


Assuntos
Ascomicetos/fisiologia , Resistência à Doença , Eczema/veterinária , Hepatopatias/veterinária , Micotoxicose/veterinária , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Doenças dos Ovinos/genética , Animais , Hepatopatias/genética , Hepatopatias/microbiologia , Micotoxicose/genética , Micotoxicose/microbiologia , Nova Zelândia , Seleção Genética , Ovinos , Doenças dos Ovinos/microbiologia , Especificidade da Espécie
6.
Animal ; 18(5): 101140, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38626708

RESUMO

Methane (CH4) is a potent gas produced by ruminants, and new measurement techniques are required to generate large datasets suitable for genetic analysis. One such technique are portable accumulation chambers (PAC), a short-term sampling method. The objectives of the current study were to explore the relationship between CH4 and carbon dioxide (CO2) output measured using both PAC and respiration chambers (RC) in growing lambs, and separately investigate the relationship among CH4, CO2 and measured ad libitum DM intake (DMI). Methane, CO2 and DMI were measured on 30 Suffolk and 30 Texel ewe lambs (age 253 ± 12 days) using the RC and PAC sequentially. The experiment was conducted over a 14-day period, with DMI measured from days 1 to 14; measurements in RC were conducted from days 10 to 12, while measurements in PAC were taken twice, the day immediately prior to the lambs entering the RC (day 9; PAC Pre-RC) and on the day lambs exited the RC (day 13; PAC Post-RC). Greater CH4 and CO2 output was measured in the RC than in the PAC (P < 0.01); similarly mean CH4 yield was greater when measured in the RC (15.39 ± 0.452 g CH4/kg DMI) compared to PAC (8.01 ± 0.767 g CH4/kg DMI). A moderate correlation of 0.37 was found between CH4 output measured in PAC Pre-RC and the RC, the corresponding regression coefficient of CH4 output measured in the RC regressed on CH4 output measured in PAC Pre-RC was close to unity (0.74; SE 0.224). The variance of CH4 and CO2 output within the measurement technique did not differ from each other (P > 0.05). Moderate to strong correlations were found between CH4 and CO2 per kg of live weight and CH4 and CO2 yield. Results from this study highlight the suitability of PAC as a ranking tool to rank animals based on their gaseous output when compared to the RC. However, repeated measurements separated by several days may be beneficial if precise rankings are required. Given the close to unity regression coefficient of CH4 output measured in the RC regressed on CH4 output measured in PAC Pre-RC suggests that PAC could also be potentially used to estimate absolute CH4 output; however, further research is required to substantiate this claim. When DMI is unknown, CH4 and CO2 per kg of live weight are a suitable alternative to the measurement of CH4 and CO2 yield.


Assuntos
Dióxido de Carbono , Gases de Efeito Estufa , Metano , Animais , Dióxido de Carbono/análise , Dióxido de Carbono/metabolismo , Metano/metabolismo , Gases de Efeito Estufa/análise , Feminino , Ovinos/fisiologia
7.
Meat Sci ; 199: 109140, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36822055

RESUMO

The inclusion of eating quality traits in sheep genetic improvement programmes is desirable. Intramuscular fat (IMF) plays a key role in ensuring consumer satisfaction when eating lamb, but genetic progress for IMF is constrained by a lack of routine data collection. This study investigated the potential for IMF predictor traits to substitute for measured IMF in genetic improvement programmes. Carcass and predicted IMF (near-infrared estimated IMF and marbling score) data were available on 10,113 New Zealand lambs, 1678 of which also had measured chemical IMF on a slice of M. longissimus lumborum on which the predictions of IMF had been made. Genetic antagonisms were observed between carcass lean traits and IMF. The genetic correlation between the predictors and measured IMF approached one, indicating that predictors of IMF can be used in genetic improvement programmes. Through using selection indexes, simultaneous increases in IMF and the existing terminal selection index are possible, provided all traits are measured. This study highlights the importance and potential of predicted IMF to achieve genetic improvement in traits of importance to consumers.


Assuntos
Músculo Esquelético , Carne Vermelha , Ovinos , Animais , Tecido Adiposo , Comportamento do Consumidor , Carne/análise
8.
Front Genet ; 13: 911355, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36186444

RESUMO

Global agreements in place to reduce methane emissions in livestock are a potential threat to food security. Successful but independent breeding strategies for improved production and lower methane are in place. The unanswered questions are whether these strategies can be combined and how they impact one another, physically and economically. The New Zealand economy is largely dependent on pastoral agriculture from grazing ruminants. The sheep industry produces ∼20 million lamb carcasses for export each year primarily from grass. Methane emitted from the fermentation of forage by grazing ruminants accounts for one-third of all New Zealand's greenhouse gas emissions. Here, we use sheep selection lines bred for divergent methane production and large numbers of their relatives to determine the genetic and phenotypic correlations between enteric methane emissions, carcass yield, and meat quality. The primary objectives were to determine whether previously shown physiological differences between methane selection lines (differing by ∼12% in methane) result in a negative impact on meat production and quality by measuring close relatives. The results show no negative effects of breeding for lowered methane on meat and carcass quality. Gross methane emissions were highly correlated with liveweight and measures of carcass weight and negatively correlated with dressing-out percentage and fat yield (GR). Trends were similar but not significant for methane yield (g CH4/kg DMI). Preliminary evidence, to date, shows that breeding for low methane may result in animals with higher lean yields that are economically favorable even before carbon costs and environmental benefits are taken into account. These benefits were seen in animals measured for methane on fixed intakes and require validation on intakes that are allowed to vary.

9.
Anim Genet ; 41(5): 449-53, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20809919

RESUMO

Until recently, the construction of a reference genome was performed using Sanger sequencing alone. The emergence of next-generation sequencing platforms now means reference genomes may incorporate sequence data generated from a range of sequencing platforms, each of which have different read length, systematic biases and mate-pair characteristics. The objective of this review is to inform the mammalian genomics community about the experimental strategy being pursued by the International Sheep Genomics Consortium (ISGC) to construct the draft reference genome of sheep (Ovis aries). Component activities such as data generation, sequence assembly and annotation are described, along with information concerning the key researchers performing the work. This aims to foster future participation from across the research community through the coordinated activities of the consortium. The review also serves as a 'marker paper' by providing information concerning the pre-publication release of the reference genome. This ensures the ISGC adheres to the framework for data sharing established at the recent Toronto International Data Release Workshop and provides guidelines for data users.


Assuntos
Genoma , Carneiro Doméstico/genética , Animais , Bovinos , Genômica/normas , Anotação de Sequência Molecular , Mapeamento Físico do Cromossomo/veterinária , Padrões de Referência
10.
Genet Mol Res ; 8(3): 1013-27, 2009 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-19731199

RESUMO

Genes whose products function in a common biological process are often co-regulated. When regulation occurs at the transcriptional level, co-expressed genes can be detected globally by expression arrays or by sequencing non-normalized cDNA libraries. We examined bovine gene expression in 27 tissues using non-normalized cDNA library sequencing. Contigs were generated from expressed sequence tags whose sequences overlapped. Contigs containing a minimum of five expressed sequence tags were ordered via a hierarchical clustering process, where the distance between the contigs represents their expression pattern similarity across tissues. Gene ontology terms associated with the genes in each cluster showed that co-clustered genes encoded proteins involved in a common biological process. This process can be used to annotate genes of unknown function in the cluster. Gene expression was compared between bovine and human tissues; there were significant correlations between species for each tissue, with the exception of thyroid and placenta. Tissues were also clustered based on the genes they express; tissues with similar physiological functions clustered closely. Based on this information, we generated the first preliminary gene atlas of the bovine genome. Genes with similar expression patterns were clustered, and genes with a common function co-clustered. This method can be used to annotate genes of unknown function in the bovine genome.


Assuntos
Bovinos/genética , Regulação da Expressão Gênica , Animais , Análise por Conglomerados , Mapeamento de Sequências Contíguas , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Biblioteca Gênica , Humanos , Filogenia , Proteínas Ribossômicas/genética
11.
Animal ; 13(3): 542-548, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30039780

RESUMO

Published studies have shown that methane yield (g CH4/kg dry matter) from sheep is positively correlated with the size (volume and surface area) of the reticulo-rumen (RR) and the weight of its contents. However, the relationship between CH4 yield and RR shape has not been investigated. In this work, shape analysis has been performed on a data set of computerised tomography (CT) scans of the RR from sheep having high and low CH4 yields (n=20 and n=17, respectively). The three-dimensional geometries of the RRs were reconstructed from segmented scan data and split into three anatomical regions. An iterative fitting technique combining radial basis functions and principal component (PC) fitting was used to create a set of consistent landmarks which were then used as variables in a PC analysis to identify shape variation within the data. Significant size differences were detected for regions corresponding to the dorsal and ventral compartments between sheep with high and low CH4 yields. When the analysis was repeated after scaling the geometries to remove the effect of size, there was no significant shape variation correlating with CH4 yield. The results have demonstrated the feasibility of CT-based computational shape determination for studying the morphological characteristics of the RR and indicate that size, but not shape correlates with CH4 yield in sheep.


Assuntos
Criação de Animais Domésticos/métodos , Dieta/veterinária , Metano/metabolismo , Rúmen/anatomia & histologia , Carneiro Doméstico/anatomia & histologia , Animais , Feminino , Seleção Genética
12.
Animal ; 13(5): 917-923, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30345952

RESUMO

Multi-sire mating of a mob of ewes is commonly used in commercial sheep production systems. However, ram mating success (defined as the number of lambs sired by an individual) can vary between rams in the mating group. If this trait was repeatable and heritable, selection of rams capable of siring larger numbers of lambs could reduce the number of rams required for mating and ultimately lead to increased genetic gain. However, genetic correlations with other productive traits, such as growth and female fertility, could influence the potential for ram mating success to be used as a selection trait. In order to investigate this trait, parentage records (including accuracy of sire assignment) from 15 commercial ram breeding flocks of various breeds were utilised to examine the repeatability and heritability of ram mating success in multi-sire mating groups. In addition, genetic and phenotypic correlations with growth and female fertility traits were estimated using ASReml. The final model used for the ram mating success traits included age of the ram and mating group as fixed effects. Older rams (3+years old) had 15% to 20% greater mating success than younger rams (1 or 2 years of age). Increasing the stringency of the criteria for inclusion of both an individual lamb, based on accuracy of sire assignment, or a whole mating group, based on how many lambs had an assigned sire, increased repeatability and heritability estimates of the ram mating success traits examined. With the most stringent criteria employed, where assignment of sire accuracy was >0.95 and the total number of lambs in the progeny group that failed to have a sire assigned was<0.05, repeatability and heritability for loge(number of lambs) was 0.40±0.09 and 0.26±0.12, respectively. For proportion of lambs sired, repeatability and heritability were both 0.30±0.09. The two ram mating traits (loge(nlamb) and proportion) were highly correlated, both phenotypically and genetically (0.88±0.01 and 0.94±0.06, respectively). Both phenotypic and genetic correlations between ram mating success and growth and other female fertility traits were low and non-significant. In conclusion, there is scope to select rams capable of producing high numbers of progeny and thus increase selection pressure on rams to increase genetic gain.


Assuntos
Hereditariedade , Reprodução/genética , Seleção Genética , Comportamento Sexual Animal , Carneiro Doméstico/fisiologia , Fatores Etários , Animais , Cruzamento , Masculino , Carneiro Doméstico/genética
13.
Dev Biol (Basel) ; 132: 205-212, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18817303

RESUMO

Gene expression was compared between resistant and susceptible Perendale lambs that had either never been exposed to gastrointestinal nematode challenge (had a naïve immune system with respect to parasites) or had been naturally challenged on pasture with nematodes. Only a small number of genes were differentially expressed between the naive resistant and susceptible animals, but many genes were differentially expressed between the resistant and susceptible challenged animals. The differentially expressed genes were involved in a variety of biological processes, most notably the immune response, the stress response and gene regulation via chromatin remodelling. The transcriptional profiling experiments also detected gene expression differences in the Ovar-DQA1 gene between resistant and susceptible challenged animals. A null allele of this gene was demonstrated to be associated with susceptibility to gastrointestinal parasites in some, but not all populations. This allele is not thought to be causal for susceptibility.


Assuntos
Perfilação da Expressão Gênica , Intestinos/parasitologia , Transcrição Gênica , Animais , Sequência de Bases , Primers do DNA , Feminino , Masculino , Ovinos
14.
Cytogenet Genome Res ; 116(1-2): 85-92, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17268183

RESUMO

Sheep chromosome 2q (OAR2q), which is homologous with human chromosome 2q (HSA2q), and cattle chromosome 2 (BTA2), is known to contain several loci contributing to carcass traits. However, the chromosomal rearrangements differentiating these chromosomes among the three species have not yet been determined and thus precise correspondences between the locations of sheep and human genes are not known. Twenty-six genes from HSA2q (2q21.1-->2q36) have been assigned to OAR2q by genetic linkage mapping to refine this area of the sheep genome. Seventy-six genes were initially selected from HSA2q. Sixty-eight percent of the PCR primer sets designed for these genes amplified successfully in sheep, and 34% amplified polymorphic products. Part of the proximal arm of OAR2q was found to be inverted compared with HSA2q. The breakpoint has been localised near the growth differentiation factor 8 gene (GDF8), spanning 380 kb between the positions of the hypothetical protein (FLJ20160) (HSA2:191008944-191075046) and glutaminase (GLS) (HSA2:191453847-191538510) (Build36.1).


Assuntos
Mapeamento Cromossômico/métodos , Animais , Bovinos , Primers do DNA/química , Bases de Dados Genéticas , Técnicas Genéticas , Genoma , Íntrons , Modelos Genéticos , Polimorfismo Genético , Ovinos
15.
Proc Biol Sci ; 274(1620): 1839-44, 2007 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-17504740

RESUMO

Livestock breeding programmes have created resistant (R) and susceptible (S) sheep that differ in their ability to control parasites through their immune function but potentially also their grazing behaviour (i.e. parasite avoidance). Using the Perendale genetic lines, we tested the hypothesis that R-sheep avoid parasites more effectively, reducing their parasite exposure/challenge, compared with S-sheep. However, in grazing systems, parasite-rich areas are also forage rich, suggesting that parasite avoidance behaviours are associated with nutritional penalties. We first created a naturally heterogeneous sward structure of gaps and tussocks and then used focal behavioural observations to quantify the sward selection of R- and S-sheep. Tussock swards were more nitrogen rich (41%), offered increased forage intake rates (32%) and contained 17 times more parasite larvae than gap swards. All the animals avoided grazing the tussock swards. However, the R-sheep grazed the tussock swards to a lesser degree than the S-sheep. We conclude that selection for genetic resistance has resulted in animals that, despite being well armed to fight parasitism through improved immune function, adopt parasite avoidance strategies with associated nutritional disadvantages. This experiment highlights the role of host behaviour in the control of parasitism and suggests that animals can be bred to avoid disease.


Assuntos
Comportamento Animal , Predisposição Genética para Doença , Doenças Parasitárias em Animais/genética , Doenças Parasitárias em Animais/parasitologia , Doenças dos Ovinos/genética , Doenças dos Ovinos/parasitologia , Animais , Ovinos , Fatores de Tempo
16.
J Anim Sci ; 95(3): 1042-1049, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28380529

RESUMO

Evidence exists from a range of species on the impact of karyotype abnormalities on reproductive performance. Despite this, cytogenetic analyses of cattle, especially females, are not routinely undertaken. Genome-wide single nucleotide polymorphism (SNP) genotype data are now, however, routinely being generated in many species globally at a relatively low cost. The objective of the present study was to evaluate the potential of routinely available SNP genotype data to identify sex-chromosome aberrations using X chromosome monosomy 59,X0 as a case study for illustration. A single 2.5-yr old Holstein-Friesian heifer was detected with a mean allelic intensity of SNP on the X chromosome almost 17 standard deviations less than the mean of other genotyped females ( = 103,326). Following cytogenetic analysis (10 replicates by karyotyping and a further 140 by FISH), the female was deduced to be a non-mosaic 59,X0. The female had never produced a calf and, although gross examination revealed no physical abnormalities, she was smaller in size than expected based on her breed and age. Given the age of the animal at slaughter, the uterus and uterine tubes appeared immature and inactive. The oviduct appeared normal while the single ovary present contained a markedly reduced number of follicles. There was, however, some evidence of prior ovulation and formation of corpora lutea. The approach proposed in the present study to identify allosome aneuploidy from routinely available genotype data can be used to screen for such abnormalities at no additional cost to the breeder or producer.


Assuntos
Doenças dos Bovinos/genética , Genótipo , Infertilidade Feminina/veterinária , Monossomia/genética , Polimorfismo de Nucleotídeo Único , Alelos , Animais , Bovinos , Feminino , Predisposição Genética para Doença , Infertilidade Feminina/genética , Cariotipagem , Gravidez
17.
Sci Rep ; 7(1): 1601, 2017 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-28487520

RESUMO

Meckel syndrome (MKS) is an inherited autosomal recessive hepatorenal fibrocystic syndrome, caused by mutations in TMEM67, characterized by occipital encephalocoele, renal cysts, hepatic fibrosis, and polydactyly. Here we describe an ovine model of MKS, with kidney and liver abnormalities, without polydactyly or occipital encephalocoele. Homozygous missense p.(Ile681Asn; Ile687Ser) mutations identified in ovine TMEM67 were pathogenic in zebrafish phenotype rescue assays. Meckelin protein was expressed in affected and unaffected kidney epithelial cells by immunoblotting, and in primary cilia of lamb kidney cyst epithelial cells by immunofluorescence. In contrast to primary cilia of relatively consistent length and morphology in unaffected kidney cells, those of affected cyst-lining cells displayed a range of short and extremely long cilia, as well as abnormal morphologies, such as bulbous regions along the axoneme. Putative cilia fragments were also consistently located within the cyst luminal contents. The abnormal ciliary phenotype was further confirmed in cultured interstitial fibroblasts from affected kidneys. These primary cilia dysmorphologies and length control defects were significantly greater in affected cells compared to unaffected controls. In conclusion, we describe abnormalities involving primary cilia length and morphology in the first reported example of a large animal model of MKS, in which we have identified TMEM67 mutations.


Assuntos
Anormalidades Múltiplas/genética , Síndrome de Dandy-Walker/genética , Síndrome Hepatorrenal/genética , Proteínas de Membrana/genética , Mutação/genética , Cisto Pancreático/genética , Anormalidades Múltiplas/patologia , Substituição de Aminoácidos , Animais , Sequência de Bases , Cromossomos de Mamíferos/genética , Cílios/patologia , Síndrome de Dandy-Walker/patologia , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Loci Gênicos , Complexo de Golgi/metabolismo , Síndrome Hepatorrenal/patologia , Homozigoto , Rim/patologia , Proteínas de Membrana/química , Mutação de Sentido Incorreto/genética , Cisto Pancreático/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ovinos , Peixe-Zebra
18.
N Z Vet J ; 64(3): 174-8, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26642120

RESUMO

AIMS: To explore and validate the utility of rumen endoscopy for collection of rumen papillae for gene expression measurement. METHODS: Four adult Coopworth ewes were fasted for either 4 or 24 hours. Animals were sedated, placed in a dorsally recumbent position at 45 degrees with the head upright, and an endoscope inserted via a tube inserted into the mouth. Biopsies of rumen papillae were taken from the ventral surface of the rumen atrium under visual guidance. Two biopsies were collected from one of the animals that had been fasted for 4 hours, and three from one of the animals that had been fasted for 24 hours. Video of the rumen atrium and reticulum was also collected. The animals recovered uneventfully. Biopsies were subsequently used for extraction and sequencing of mRNA. RESULTS: The ventral surface of the rumen atrium was accessible after 4 hours off pasture, but a larger region was accessible after 24 hours of fasting. Sedation allowed access for endoscope use for around 5 to 10 minutes after which increased saliva flow was noted. Rumen papillae biopsies were easily collected, with samples from a variety of sites collected in the ∼10 minute time window. High quality RNA was obtained for stranded mRNA sequencing. Of the resulting reads, 69-70% mapped uniquely to version 3.1 of the ovine genome, and 48-49% to a known gene. The rumen mRNA profiles were consistent with a previously reported study. CONCLUSIONS: This method for obtaining rumenal tissue was found to be rapid and resulted in no apparent short or long term effects on the animal. High quality RNA was successfully extracted and amplified from the rumen papillae biopsies, indicating that this technique could be used for future gene expression studies. The use of rumen endoscopy could be extended to collection of a variety of rumen and reticulum anatomical measurements and deposition and retrieval of small sensors from the rumen. Rumen endoscopy offers an attractive and cost effective approach to repeated rumen biopsies compared with serial slaughter or use of cannulated animals.


Assuntos
Endoscopia Gastrointestinal/veterinária , Rúmen/patologia , Ovinos , Animais , Biópsia/métodos , Biópsia/veterinária , Endoscopia Gastrointestinal/métodos , Feminino
19.
Genetics ; 160(3): 1113-22, 2002 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11901127

RESUMO

The last decade has seen a dramatic increase in the number of livestock QTL mapping studies. The next challenge awaiting livestock geneticists is to determine the actual genes responsible for variation of economically important traits. With the advent of high density single nucleotide polymorphism (SNP) maps, it may be possible to fine map genes by exploiting linkage disequilibrium between genes of interest and adjacent markers. However, the extent of linkage disequilibrium (LD) is generally unknown for livestock populations. In this article microsatellite genotype data are used to assess the extent of LD in two populations of domestic sheep. High levels of LD were found to extend for tens of centimorgans and declined as a function of marker distance. However, LD was also frequently observed between unlinked markers. The prospects for LD mapping in livestock appear encouraging provided that type I error can be minimized. Properties of the multiallelic LD coefficient D' were also explored. D' was found to be significantly related to marker heterozygosity, although the relationship did not appear to unduly influence the overall conclusions. Of potentially greater concern was the observation that D' may be skewed when rare alleles are present. It is recommended that the statistical significance of LD is used in conjunction with coefficients such as D' to determine the true extent of LD.


Assuntos
Desequilíbrio de Ligação , Característica Quantitativa Herdável , Ovinos/genética , Animais , Marcadores Genéticos , Haplótipos , Heterozigoto , Repetições de Microssatélites , Modelos Genéticos
20.
Meat Sci ; 71(2): 383-91, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22064240

RESUMO

Comparisons were made of carcass and meat quality characteristics of pasture-raised Texel-cross ewe (n=269) and ram (n=275) lambs between 5 and 8 months of age with an average carcass weight of 17.2kg. Carcass assessment was based on linear dimensions and dissection of a leg into muscle, fat and bone, and meat quality measurements were made on M. semimembranosus and M. longissimus. At a set carcass weight, ewe lambs had higher dressing percentages (2%), shorter carcasses (0.7cm), and heavier leg cuts (35g) (P<0.01) than males. At the same leg weight, legs of ewe lambs were fatter than males (subcutaneous plus intermuscular fat; 11.2% vs 9.6%; P<0.001), whereas legs of ram lambs contained significantly more muscle and bone than females (P<0.001). Leg muscle to bone ratio (4.7 vs 4.4) and muscularity were higher for females than males (P<0.001). However, the relationship between leg muscle to bone ratio and muscularity was not the same between the sexes, and for any given muscularity value the muscle to bone ratio of ram lambs was lower than females. As a result, if carcass lean meat yield is predicted from a measure of carcass shape, such as muscularity, lean meat yields will be overestimated for males and underestimated for females. Meat quality was lower in ram lambs than in females (P<0.001) as shown by higher Warner-Bratzler shear values (peak value 109.8 vs 97.0N for M. Semimembranosus), higher ultimate meat pH values, and lower redness (a(∗)) and lightness (L(∗), for the longissimus muscle only) values (P<0.001). It is concluded that significant differences between ewe and ram lambs do exist for many carcass and meat quality traits, but for most quality traits the differences are small.

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