Why do physicians go to work when they are sick? Presenteeism at different career stages.

INTRODUCTION AND OBJECTIVE: Physicians find it difficult to take on the role of the patient and they show unusual behaviors when ill. One of these behaviors is presenteeism, which is working while sick. The objective of this research is to analyze the factors that contribute to the phenomenon of presenteeism in Spanish physicians. MATERIAL AND METHODS: Mixed methodology study: one national survey through the General Council of Medical Associations website (quantitative part), 22 semistructured interviews with sick residents and practicing physicians, and three focus groups involving professionals from the occupational health services (qualitative). A bivariate analysis using parametric and non-parametric tests. The significance level was p<0.05 (95% confidence interval). Qualitative analysis using the comparative-constant method until saturation of information. RESULTS: Presenteeism is reported by 89.4% of doctors who responded to the survey, and it is more common among women. Contributing factors include fear of overburdening colleagues (the main reason and more common among women 58.14% vs 48.35%), self-perception of doing one's duty (the second reason and more common among men, 44.63% vs 33.14%) and economic impact and difficulty in accepting the role of a sick person. This behavior has an impact on patient safety, and is part of the hidden curriculum that also affects the training of medical professionals. CONCLUSIONS: Presenteeism is a widespread and accepted practice among medical professionals. Although normalized, and even appreciated as a way to avoid overburdening colleagues, presenteeism has important implications for clinical ethics and patient safety.

Gender analysis of Spanish National Questionnaire on behaviours and attitudes of doctors towards their own illness (CAMAPE).

INTRODUCTION AND OBJECTIVES: Physicians' health is a key element for quality healthcare. Medical professionals have difficulty accepting their role as patients and it might be different among sexes. The aim was to describe behaviours and attitudes of doctors towards their own illness. MATERIALS AND METHODS: An online survey was launched through the General Council of Medical Associations webpage for all Spanish registered doctors. A bivariate analysis by sex was performed for all the questionnaire variables using parametric and non-parametric tests. The significance level was p<0.05 (95% confidence interval). RESULTS: A total of 4,308 registered doctors (1,858 men and 2,450 women) answered. Women were younger, single, and worked mainly in non-surgical specialities in the public sector. Men were older, married, and worked more frequently in public-private practice. Women had less chronic conditions, except for anxiety disorders (11.52% vs 15.18%). Both sexes, especially women, primarily self-treated (94.29% vs 95.02%), went to work while ill (88.16% vs 90.29%), visited their GP (56% vs 70%), and half of them underwent annual occupational health checks (40% vs 48%). Women self-prescribed more analgesics (93.43% vs 95.63%), more presenteeism (88% vs 90%) and felt more insecure when treating sick fellows (9.96% vs 20.12%) and requested training for it. More women agreed to make deontological recommendations about doctors' health (91.55% vs 96.16%) and considered revalidation may contribute to improve doctors' health (65.29% vs 66.16%). CONCLUSIONS: Male and female doctors show illness-health behaviours and attitudes at work to improve. There are differences among male and female doctors. Regarding, medical feminization, ethical recommendations may be of benefit regarding doctors' health-illness issues and considering gender perspective.

Behaviour and attitudes of Spanish physicians towards their own process of falling ill: Study protocol and validation of CAMAPE questionnaire.

Physicians have not learned their role as patients. Health programmes for doctors are focused on mental health. Nevertheless, anomalous behaviours of ill doctors exist independently of health problems. We present a study to describe behaviour and attitudes of doctors towards their own illness (CAMAPE) including the analysis of questionnaire validation. MATERIAL AND METHODS: A mix methodology study based on semi-structured interviews to ill physicians and focus groups with members of medical colleges, occupational medicine services and doctors of ill doctors was performed. A survey was designed. Survey validation process included content and face validity, construct validity through exploratory and confirmatory factor analysis and reliability by Cronbach's Alpha Index. RESULTS: A total of 27 interviews to ill doctors and 4 focus group were performed. Content and feasibility assessment was made by experts. Psychometric validation was performed with a sample of 4308 answers (2450 women, 56.87%). A 5-factor (F) model explained 78.08% variance. First factor (F1) "The work might worsen health". Second (F2) "Mental issues, toxic habits and the impact of a bad health on work performance"; Third (F3) presenteeism and sick leaves; Fourth (F4) the handling of an ill colleague and the role of medical colleges. Fifth (F5) the healthcare pathway and potential value of revalidation in medical profession. CONCLUSIONS: A comprehensive mixed study on the process of physicians becoming ill has been launched with a reliable questionnaire in a large sample of registered doctors. The analysis will help to formulate gender-sensitive policy and ethical recommendations in relation to sick doctors given the progressive feminisation of the medical profession.

SNP rs403212791 in exon 2 of the MTNR1A gene is associated with reproductive seasonality in the Rasa aragonesa sheep breed.

The aim of this study was to characterize and identify causative SNPs in the MTNR1A gene responsible for the reproductive seasonality traits in the Rasa aragonesa sheep breed. A total of 290 ewes (155, 84 and 51 mature, young and ewe lambs, respectively) from one flock were controlled from January to August. The following three reproductive seasonality traits were considered: the total days of anoestrus (TDA) and the progesterone cycling months (P4CM); both ovarian function seasonality traits based on blood progesterone levels; and the oestrus cycling months (OCM) based on oestrous detection, which indicate behavioural signs of oestrous. We have sequenced the total coding region plus 733 and 251 bp from the promoter and 3'-UTR regions, respectively, from the gene in 268 ewes. We found 9 and 4 SNPs associated with seasonality traits in the promoter (for TDA and P4CM) and exon 2 (for the three traits), respectively. The SNPs located in the gene promoter modify the putative binding sites for various trans-acting factors. In exon 2, two synonymous SNPs affect RFLP sites, rs406779174/RsaI (for the three traits) and rs430181568/MnlI (for OCM), and they have been related with seasonal reproductive activity in previous association studies with other breeds. SNP rs400830807, which is located in the 3'-UTR, was associated with the three traits, but this did not modify the putative target sites for ovine miRNAs according to in silico predictions. Finally, the SNP rs403212791 (NW_014639035.1: g.15099004G > A), which is also associated with the three seasonality phenotypes, was the most significant SNP detected in this study and was a non-synonymous polymorphism, leading a change from an Arginine to a Cysteine (R336C). Haplotype analyses confirmed the association results and showed that the effects found for the seasonality traits were caused by the SNPs located in exon 2. We have demonstrated that the T allele in the SNP rs403212791 in the MNTR1A gene is associated with a lower TDA and higher P4CM and OCM values in the Rasa Aragonesa breed.

Efficacy of a global supportive skin care programme with hydrotherapy after non-metastatic breast cancer treatment: A randomised, controlled study.

This study investigated the efficacy of post-treatment hydrotherapy as supportive care for management of persistent/long-lasting dermatologic adverse events (dAEs) induced in breast cancer survivors by adjuvant therapy, and its impact on quality of life (QoL). Patients in complete remission after standardised (neo)adjuvant chemotherapy, surgery and radiotherapy combination treatment for infiltrating HR+/HER2-breast carcinoma were enrolled in this randomised, multicentre controlled study 1-5 weeks after completing radiotherapy. The control group (CG, n = 33) received best supportive care and the treatment group (HG, n = 35) received 3-weeks of specific hydrotherapy. The primary criterion was change in QoL (QLQ-BR23) after hydrotherapy. Clinical grading of dAEs, cancer-related QoL (QLQ-C30), dermatologic QoL (DLQI) and general psychological well-being (PGWBI) were assessed. Significant dAEs were found at inclusion in both groups (n = 261). Most items showed significantly greater improvement in the HG versus CG group: QLQ-BR23 (breast [p = .0001] and arm symptoms [p = .0015], systemic therapy side effects [p = .0044], body image [p = .0139]), some dAE grading, DLQI (p = .0002) and PGWBI (p = .0028). Xerosis (88% of patients at inclusion) completely healed in all HG patients. Specific hydrotherapy is an effective supportive care for highly prevalent and long-lasting dAEs occurring after early breast cancer treatment, including chemotherapy, and leads to improved QoL and dermatologic toxicities.

Feature domain-specific movement intention detection for stroke rehabilitation with brain-computer interfaces.

Brain-computer interface (BCI) driven electrical stimulation has been proposed for neuromodulation for stroke rehabilitation by pairing intentions to move with somatosensory feedback from electrical stimulation. Movement intentions have been detected in several studies using different techniques, with temporal and spectral features being the most common. A few studies have compared temporal and spectral features, but conflicting results have been reported. In this study, the aim was to investigate if complexity measures can be used for movement intention detection and to compare the detection performance based on features extracted from three different domains (time, frequency and complexity) from single-trial EEG. Two data sets were used where four different isometric palmar grasps or dorsiflexions were performed while continuous EEG was recorded. 39 healthy subjects performed or imagined these movements and 11 stroke patients attempted to perform the movements. The EEG was pre-processed and divided into two epoch classes: Background EEG (2 s) and movement intention (2 s). To obtain an estimated detection performance, temporal, spectral and complexity features were extracted and classified (linear discriminant analysis) after the feature vector was reduced using sequential forward selection. The results show that accuracies between 82-87% and 74-80% are obtained for foot and hand movements, respectively. The temporal feature domain was the most dominant for foot movement intention detection, while the spectral features contributed more to the hand movement detection. The complexity features could be used to detect movement intentions, but the performance was much lower compared to temporal and spectral features.

Parálisis del tercer nervio craneal y retinopatía de Purtscher en niña politraumatizada / Third cranial nerve palsy and Purtscher retinopathy in a child with multiple injuries

CASO CLÍNICO: Una niña de 4 años ingresa por politraumatismo grave por arrollamiento. Durante la evolución se le diagnostica de parálisis completa del tercer par del ojo derecho y retinopatía de Purtscher en ojo izquierdo. DISCUSIÓN: La retinopatía de Purtscher es poco frecuente. El diagnóstico es clínico y su tratamiento no está estandarizado aunque los corticoides sistémicos podrían mejorar el pronóstico visual. Las parálisis deL tercer par traumáticas tienen baja tendencia a la recuperación espontánea. El uso de toxina botulínica en niños podría mejorar la tasa de recuperación total y podría ser útil para acelerar la resolución y así permitir binocularidad y evitar la ambliopía

CASE REPORT: A 4 year-old girl was referred to our hospital after have suffered a severe accident. The patient was diagnosed with complete third nerve palsy in her right eye and Purtscher retinopathy in her left eye. DISCUSSION: Purtscher retinopathy is a rare condition. The diagnosis is made on clinical ground and its treatment is not well defined although it is believed that systemic steroids could improve the visual outcome. Traumatic third nerve palsy has a poor spontaneous recovery. The use of botulinum toxin might be useful in children to improve the recovery rate, maintaining binocularity, and avoiding amblyopia in other cases

[Third cranial nerve palsy and Purtscher retinopathy in a child with multiple injuries]. / Parálisis del tercer nervio craneal y retinopatía de Purtscher en niña politraumatizada.

CASE REPORT: A 4 year-old girl was referred to our hospital after have suffered a severe accident. The patient was diagnosed with complete third nerve palsy in her right eye and Purtscher retinopathy in her left eye. DISCUSSION: Purtscher retinopathy is a rare condition. The diagnosis is made on clinical ground and its treatment is not well defined although it is believed that systemic steroids could improve the visual outcome. Traumatic third nerve palsy has a poor spontaneous recovery. The use of botulinum toxin might be useful in children to improve the recovery rate, maintaining binocularity, and avoiding amblyopia in other cases.

Characterisation of the Melatonin Receptor 1A (MTNR1A) gene in the Rasa Aragonesa sheep breed: association with reproductive seasonality.

The ovine Melatonin Receptor 1A (MTNR1A) gene was structurally characterised and association between its variants and the reproductive seasonality was examined in a daughter design comprising three families of Rasa Aragonesa sheep breed. Sequencing of six Rasa Aragonesa ewes with extreme values for seasonality trait revealed 28 polymorphisms: 11 SNPs in the coding region (all in Exon 2), and 17 SNPs in the promoter region MTNR1A. All the substitutions in the coding region were found most likely lacking any phenotypic effect, because they are conservative mutations or were not part of the transmembrane domain. The silent mutations, which had shown association with reproductive seasonality in other breeds, were also found and genotyped in Rasa Aragonesa. The T allele of SNP606/RsaI of MNTR1A gene was associated with a greater percentage of oestrous cyclic ewes in the Rasa Aragonesa breed, indicating that this SNP may be in linkage disequilibrium with a mutation responsible for this trait close to MTNR1A, or in regulatory sequences of the gene. In this sense, several SNPs affecting a binding element for some transcription factors have been identified in the promoter region. The SNPs at 422 and 527 positions could constitute a binding element for some transcription factors (TFs), located in an EF2 and SRY consensus sites in the promoter region, respectively. Haplotype h(5) showed significant differences with the h(2) haplotype (66% compared to 49.2%) on oestrous cyclicity, thus these results are consistent with genotypic associations for each SNP. Haplotype with T, A and T alleles for SNPs 422, 677 (promoter region) and 612 (Exon 2) showed an increase of the percentage of oestrous cyclic ewes. Although some of these mutations have been associated with seasonal reproduction, further studies with a more appropriate animal design as well as functional studies of TF binding activity are needed.

Effect of the FecX(R) polymorphism in the bone morphogenetic protein 15 gene on natural or equine chorionic gonadotropin-induced ovulation rate and litter size in Rasa Aragonesa ewes and implications for on-farm application.

A new mutation in the bone morphogenetic protein 15 (BMP15) gene (FecX(R) allele) causing increased prolificacy in heterozygous (R+) and sterility in homozygous ewes has been recently described in Rasa Aragonesa, a low-prolificacy Mediterranean breed. The current study determined, first, the effect of this polymorphism on natural and eCG-induced ovulation rate (OR) and the effect of eCG dose on reproductive performance; and second, its effect on prolificacy and its interaction with progestagen + eCG treatment on farms, which have not been reported to date. The FecX(R) allele increased OR by 0.44 and 0.63 ovulations in young (n = 91) and adult (n = 84) R+ ewes, respectively (both, P < 0.01), increments less than reported in prolific breeds carrying other mutations in BMP15. When the standard dose of eCG used on farms (480 IU) was applied to R+ ewes (n = 36), an extremely high OR (3.95) was recorded, which was accompanied by greater partial failure of multiple ovulations (PFMO). On the contrary, OR using 240 IU in R+ ewes (2.90; n = 35) was similar to 480 IU in wildtype (++) ewes (2.82; n = 48; both P < 0.01 when compared with 480 IU in R+ ewes). No differences were found in the birth weight of the offspring between R+ and ++ eCG-stimulated ewes within the same litter size. To validate the genealogy identification on farms, PCR genotyping was carried out in 1,667 ewes from 4 elite flocks, resulting in a negligible misclassification of R+ ewes, which demonstrated that identification by genealogy is a reliable tool to identify FecX(R) ewes within the breeding program. In recorded farms, the natural litter size of ++ ewes (1.34, n = 599,160 lambing records) was increased due to the FecX(R) allele by 0.35 lambs (P < 0.0001, n = 6,593 lambing records). A similar increase (0.30) was observed when comparing ++ and R+ ewes treated with 480 IU of eCG (P < 0.0001, n = 62,055 and n = 866, respectively). When applying 480 IU of eCG to R+ ewes, the increase in prolificacy was only due to increased percentages of triplets (P < 0.001) and quadruplets (P < 0.0001), but not of twin births. In conclusion, the favorable reproductive performance of R+ ewes, with 0.63 extra ovulations and 0.35 extra lambs per lambing ewe, is responsible for the increased interest in the use of this polymorphism. Nevertheless, care must be taken in the application of eCG to R+ ewes, with the current results showing that the standard dose increases prolificacy by only increasing triple and higher-order births.

[Clinical pathways for the care of multiple sclerosis patients]. / Diseño de una vía clínica para la atención a los pacientes con esclerosis múltiple.

INTRODUCTION: clinical pathways are standard health care methods to coordinate clinical work, reduce inter-clinician variability, improve patient care and increase staff and patient satisfaction. The aim of this study is to develop a clinical pathway capable of organising and developing standard procedures for diagnosis, treatment and care in patients with multiple sclerosis and to coordinate all medical specialists involved in this disease. METHODS: a multidisciplinary unit for the care of MS patients was developed. All of them and quality specialists analysed some international evidence-based studies, clinical guides, international guidelines and other clinical neurological pathways in several meetings and designed several documents for the clinical pathways. RESULTS: a clinical pathway was created consisting of a scientific-technical framework, which arranges the care in relation to the diagnosis and reatment. The framework is accompanied by various patient-information documents on the disease, an information sheet on diagnostic procedures and a map of the process. Quality standards were established to achieve continuous improvement in patient care. CONCLUSIONS: a clinical pathway for the care of MS patients in a multidisciplinary unit homogenises and organises the care which the MSpatient should receive from the initial symptoms to the progressive stages. This clinical pathway improves the quality of patient care, reduces the variability in work protocols and rationalises the use of the available health care resources.

Diseño de una vía clínica para la atención a los pacientes con esclerosis múltiple / Clinical pathways for the care of multiple sclerosis patients

Introducción: Las vías clínicas (VC) son herramientas para coordinar el trabajo asistencial, reducir la variabilidad entre el personal sanitario y mejorar la atención y el cuidado del paciente. La esclerosis múltiple (EM) es una enfermedad neurológica crónica que afecta a pacientes jóvenes y es incapacitante. El objetivo es desarrollar una vía clínica para mejorar el diagnóstico, el tratamiento y la atención de los pacientes con EM y, asimismo, facilitar la coordinación de todos los especialistas implicados en este proceso. Método: Siguiendo el modelo FOCUS-PDCA se organiza un equipo de trabajo integrado por diferentes profesionales implicados en la atención del paciente con EM. Se realiza una revisión bibliográfica exhaustiva y se llega a consenso; así, se diseñan los documentos de la VC con base en la evidencia científica. Resultados: Se crea una vía clínica compuesta por los siguientes elementos: una matriz temporal con una serie de anexos para ordenar el proceso diagnóstico y el tratamiento, un impreso de información a los pacientes sobre las pruebas diagnósticas, una hoja de información sobre la enfermedad y un mapa de procesos, una encuesta de evaluación de la calidad percibida y un documento con indicadores de calidad para evaluar la VC. Conclusiones: El desarrollo de una VC de EM facilita la atención multidisciplinaria y mejora la calidad asistencial. Esta propuesta es novedosa al enfocar la atención integral de la EM desde su inicio, tanto en aspectos diagnósticos como terapéuticos, incluyendo el ámbito ambulatorio (AU)

Introduction: Clinical pathways are standard health care methods to coordinate clinical work, reduce inter-clinician variability, improve patient care and increase staff and patient satisfaction. The aim of this study is to develop a clinical pathway capable of organising and developing standard procedures for diagnosis, treatment and care in patients with multiple sclerosis and to coordinate all medical specialists involved in this disease. Methods: A multidisciplinary unit for the care of MS patients was developed. All of them and quality specialists analysed some international evidence-based studies, clinical guides, international guidelines and other clinical neurological pathways in several meetings and designed several documents for the clinical pathways. Results: A clinical pathway was created consisting of a scientific-technical framework, which arranges the care in relation to the diagnosis and reatment. The framework is accompanied by various patient-information documents on the disease, an information sheet on diagnostic procedures and a map of the process. Quality standards were established to achieve continuous improvement in patient care. Conclusions: A clinical pathway for the care of MS patients in a multidisciplinary unit homogenises and organises the care which the MSpatient should receive from the initial symptoms to the progressive stages. This clinical pathway improves the quality of patient care, reduces the variability in work protocols and rationalises the use of the available health care resources (AU)

The bovine annexin 9 gene (ANXA9) is significantly associated with milk-fat yield in a Spanish Holstein-Friesian population.

On the basis of QTL studies for milk-fat yield trait on BTA3, annexin 9 protein (ANXA9), fatty acid transport protein type 3 (SLC27A3) and diacylglycerol O-acyltransferase 1 (DGAT1) were selected as candidate genes. Three different single nucleotide polymorphisms (SNPs) of bovine ANXA9, SLC27A3 and DGAT1 genes have been tested in a selective genotyping design for milk-fat yield. Significant allele frequency differences were found for ANXA9 (p=0.02), in Holstein-Friesian animals with high and low breeding values for milk-fat yield. Regression analysis also showed a significant effect (p=0.0207) between estimated breeding values (EBVs) for fat milk content and ANXA9 polymorphism. So ANXA9 gene falls into a significant quantitative trait loci interval for milk-fat yield that was previously reported on bovine chromosome 3 in other dairy populations. Our results suggest that the ANXA9 gene polymorphism or a linked segregating QTL contributes to variation in milk-fat yield.

Gestación ectópica cervical tratada mediante punción ecoguiada intrasacular de metotrexato / Cervical pregnancy treated with local injection of methotrexate guided by transvaginal ultrasonography

El embarazo cervical es una forma infrecuente de gestación ectópica. El examen ecográfico permite el diagnóstico precoz, y una actuación conservadora. Se describe el caso clínico de una gestación ectópica cervical diagnosticada en el Servicio de Urgencias de Obstetricia y Ginecología del Hospital Universitario Miguel Servet (Zaragoza, España). Paciente nulípara de 31 años que consultó por sangrado genital. El diagnóstico se realizó mediante ecografía transvaginal, objetivándose un embarazo ectópico cervical con embrión vivo acorde a 8 semanas de edad gestacional. Se realizó tratamiento conservador con metotrexato mediante inyección local intrasacular asociado a administración sistémica. Seguimiento con determinaciones seriadas de gonodotrópica coriónica humana, objetivándose un descenso progresivo de ésta. Se efectuaron controles ecográfcos periódicos, evidenciando la reabsorción de la gestación confirmándose el éxito del tratamiento.

Cervical ectopic pregnancy is an unusual form of ectopic pregnancy. Ultrasound examination makes early diagnosis possible and permits to perform conservative treatment. We describe a case of cervical ectopic pregnancy diagnosed in the Emergency Section of Miguel Servet University Hospital (Zaragoza, Spain). A 31 year-old nullipara pregnant woman came to our Hospital because of genital bleeding. Ultrasound trans-vaginal examination demonstrated a cervical pregnancy with an 8 weeks embryo. Conservative treatment was prescribed using US-guided injection and systemic methotrexate. After treatment, weekly quantitative human chorionic gonadotrophin levels were determined and decreased progressively. Serial ultrasound exams were performed demonstrating how pregnancy products underwent resorption and confirming therapy success.

Variations in the mitochondrial cytochrome c oxidase subunit I gene indicate northward expanding populations of Culicoides imicola in Spain.

Culicoides imicola is the main vector for bluetongue (BT) and African horse sickness (AHS) viruses in the Mediterranean basin and in southern Europe. In this study, we analysed partial mitochondrial cytochrome c oxidase subunit I (COI) gene to characterize and confirm population expansion of Culicoides imicola across Spain. The data were analysed at two hierarchical levels to test the relationship between C. imicola haplotypes in Spain (n = 215 from 58 different locations) and worldwide (n = 277). We found nineteen different haplotypes within the Spanish population, including 11 new haplotypes. No matrilineal subdivision was found within the Spanish population, while western and eastern Mediterranean C. imicola populations were very structured. These findings were further supported by median networks and mismatch haplotype distributions. Median networks demonstrated that the haplotypes we observed in the western Mediterranean region were closely related with one another, creating a clear star-like phylogeny separated only by a single mutation from eastern haplotypes. The two, genetically distinct, sources of C. imicola in the Mediterranean basin, thus, were confirmed. This type of star-like population structure centred around the most frequent haplotype is best explained by rapid expansion. Furthermore, the proposed northern expansion was also supported by the statistically negative Tajima's D and Fu's Fs values, as well as predicted mismatch distributions of sudden and spatially expanding populations. Our results thus indicated that C. imicola population expansion was a rapid and recent phenomenon.

Freemartinism and FecXR allele determination in replacement ewes of the Rasa Aragonesa sheep breed by duplex PCR.

A new naturally occurring mutation in the fecundity gene BMP15 in the Rasa Aragonesa sheep breed (Ovis aries) has been found to affect prolificacy. This mutation (FecX(R) allele) is a deletion of 17 base pairs that leads to an altered amino acid sequence, and this alteration increases prolificacy in heterozygous ewes but causes sterility in homozygous ewes. Selection of repository lambs with the FecX(R) allele increases rates of twins and multiple lambing and thereby also increases the probability of lambing freemartins that will become sterile. In this sense, an accurate, reliable, and quick method was developed by duplex polymerase chain reaction (PCR) for sex, amplifying an ovine-specific Y chromosome repetitive fragment, and BMP15 genotype determination in replacement ewe lambs. The BMP15 fragment served as an internal control of the amplification and detected the FecX(R) allele, avoiding a false negative and then a mistake in freemartin detection. This assay uncovered 6 freemartin females among 195 replacement ewes from 7 different commercial flocks and 1 experimental flock. Furthermore, 1554 rams from 64 commercial flocks were also analyzed to identify FecX(R) rams. This analysis identified 103 rams hemizygous for the FecX(R) allele and 1 heterozygous ram. Because this gene is located on the X chromosome, this heterozygous animal is a freemartin ram that is co-amplifying the DNA from XX and XY lymphocytes. These results confirm the usefulness of this multiplex PCR assay for detecting phenotypically sexed females, freemartins, and the BMP15 genotype to detect highly prolific ewes in commercial flocks and to assist breeders in selection of repository lambs.

Reliability of sex determination in ovine embryos using amelogenin gene (AMEL).

Sex specific sequence variability of the amelogenin gene has been used for sex determination in the family of Bovidae. In our study, suitability and reliability of the amelogenin gene for ovine sex determination in embryos was studied. The specificity of the method was previously demonstrated by testing 579 blood samples from several Spanish sheep breeds (161 males and 198 females). No amplification failures and very high agreement between genotypic and phenotypic sex was found (578/579), in contrast to humans, where errors in sex determination has been reported because of mutations in AMELX or AMELY genes. However, one female animal showed a male genotypic sex, being the most plausible explanation that a recombination event has happened during the meiosis. In our study only 0.17% (1/579) of the samples tested has been misdiagnosed using the amelogenin gene. Finally, 1-10 cells from each of 67 compact morulae were aspirated through the zona pellucida, and genotyped for sex determination. The efficiency in sex determination was 95 and 98% when more than two and more than three cells were sampled, respectively. The total time required for the genetic test, was less than 4h. These results confirm that the amelogenin gene can be used for rapid sex determination in ovine embryos, with a high efficiency and accuracy (100%) when three or more cells are sampled, allowing transferring sexed fresh embryos in MOET programmes. To our knowledge, this was the first time that sex determination using the amelogenin gene was performed in ovine embryos.

A deletion in the bone morphogenetic protein 15 gene causes sterility and increased prolificacy in Rasa Aragonesa sheep.

Bone morphogenetic protein 15 (BMP15) is a member of the transforming growth factor beta superfamily, is specifically expressed in oocytes and is essential for sheep prolificacy. Reported mutations in this gene cause increased ovulation rate and infertility in a dosage-sensitive manner. In this work, a new naturally occurring mutation in the BMP15 gene from the ovine Rasa Aragonesa breed is described. This mutation is a deletion of 17 bp that leads to an altered amino acid sequence and introduces a premature stop codon in the protein. Highly significant associations (P < 0.0001) were found between the estimated breeding value for prolificacy and the genotype of BMP15 in Rasa Aragonesa animals with high and low breeding values for this trait. As for other mutations in BMP15, this new mutation is associated with increased prolificacy and sterility in heterozygous and homozygous ewes respectively.