DNA-based diagnosis of rare diseases in veterinary medicine: a 4.4 kb deletion of ITGB4 is associated with epidermolysis bullosa in Charolais cattle.

BACKGROUND Rare diseases in livestock animals are traditionally poorly diagnosed. Other than clinical description and pathological examination, the underlying causes have, for the most part, remained unknown. A single case of congenital skin fragility in cattle was observed, necropsy, histological and ultrastructural examinations were carried out and whole genome sequencing was utilized to identify the causative mutation. RESULTS A single purebred female Charolais calf with severe skin lesions was delivered full-term and died spontaneously after birth. The clinical and pathological findings exactly matched the gross description given by previous reports on epitheliogenesis imperfecta and epidermolysis bullosa (EB) in cattle. Histological and ultrastructural changes were consistent with EB junctionalis (EBJ). Genetic analysis revealed a previously unpublished ITGB4 loss-of-function mutation; the affected calf was homozygous for a 4.4 kb deletion involving exons 17 to 22, and the dam carried a single copy of the deletion indicating recessive inheritance. The homozygous mutant genotype did not occur in healthy controls of various breeds but some heterozygous carriers were found among Charolais cattle belonging to the affected herd. The mutant allele was absent in a representative sample of unrelated sires of the German Charolais population. CONCLUSION This is the first time in which a recessively inherited ITGB4 associated EBJ has been reported in cattle. The identification of heterozygous carriers is of importance in avoiding the transmission of this defect in future. Current DNA sequencing methods offer a powerful tool for understanding the genetic background of rare diseases in domestic animals having a reference genome sequence available.

Organ Culture Bioreactors - Platforms to Study Human Intervertebral Disc Degeneration and Regenerative Therapy.

In recent decades the application of bioreactors has revolutionized the concept of culturing tissues and organs that require mechanical loading. In intervertebral disc (IVD) research, collaborative efforts of biomedical engineering, biology and mechatronics have led to the innovation of new loading devices that can maintain viable IVD organ explants from large animals and human cadavers in precisely defined nutritional and mechanical environments over extended culture periods. Particularly in spine and IVD research, these organ culture models offer appealing alternatives, as large bipedal animal models with naturally occurring IVD degeneration and a genetic background similar to the human condition do not exist. Latest research has demonstrated important concepts including the potential of homing of mesenchymal stem cells to nutritionally or mechanically stressed IVDs, and the regenerative potential of "smart" biomaterials for nucleus pulposus or annulus fibrosus repair. In this review, we summarize the current knowledge about cell therapy, injection of cytokines and short peptides to rescue the degenerating IVD. We further stress that most bioreactor systems simplify the real in vivo conditions providing a useful proof of concept. Limitations are that certain aspects of the immune host response and pain assessments cannot be addressed with ex vivo systems. Coccygeal animal disc models are commonly used because of their availability and similarity to human IVDs. Although in vitro loading environments are not identical to the human in vivo situation, 3D ex vivo organ culture models of large animal coccygeal and human lumbar IVDs should be seen as valid alternatives for screening and feasibility testing to augment existing small animal, large animal, and human clinical trial experiments.

Novel Features of the Prenatal Horn Bud Development in Cattle (Bos taurus).

Whereas the genetic background of horn growth in cattle has been studied extensively, little is known about the morphological changes in the developing fetal horn bud. In this study we histologically analyzed the development of horn buds of bovine fetuses between ~70 and ~268 days of pregnancy and compared them with biopsies taken from the frontal skin of the same fetuses. In addition we compared the samples from the wild type (horned) fetuses with samples taken from the horn bud region of age-matched genetically hornless (polled) fetuses. In summary, the horn bud with multiple layers of vacuolated keratinocytes is histologically visible early in fetal life already at around day 70 of gestation and can be easily differentiated from the much thinner epidermis of the frontal skin. However, at the gestation day (gd) 212 the epidermis above the horn bud shows a similar morphology to the epidermis of the frontal skin and the outstanding layers of vacuolated keratinocytes have disappeared. Immature hair follicles are seen in the frontal skin at gd 115 whereas hair follicles below the horn bud are not present until gd 155. Interestingly, thick nerve bundles appear in the dermis below the horn bud at gd 115. These nerve fibers grow in size over time and are prominent shortly before birth. Prominent nerve bundles are not present in the frontal skin of wild type or in polled fetuses at any time, indicating that the horn bud is a very sensitive area. The samples from the horn bud region from polled fetuses are histologically equivalent to samples taken from the frontal skin in horned species. This is the first study that presents unique histological data on bovine prenatal horn bud differentiation at different developmental stages which creates knowledge for a better understanding of recent molecular findings.

Hybrid ‘superswarm’ leads to rapid divergence and establishment of populations during a biological invasion

Understanding the genetic background of invading species can be crucial information clarifying why they become invasive. Intraspecific genetic admixture among lineages separated in the native ranges may promote the rate and extent of an invasion by substantially increasing standing genetic variation. Here we examine the genetic relationships among threespine stickleback that recently colonized Switzerland. This invasion results from several distinct genetic lineages that colonized multiple locations and have since undergone range expansions, where they coexist and admix in parts of their range. Using 17 microsatellites genotyped for 634 individuals collected from 17 Swiss and two non-Swiss European sites, we reconstruct the invasion of stickleback and investigate the potential and extent of admixture and hybridization among the colonizing lineages from a population genetic perspective. Specifically we test for an increase in standing genetic variation in populations where multiple lineages coexist. We find strong evidence of massive hybridization early on, followed by what appears to be recent increased genetic isolation and the formation of several new genetically distinguishable populations, consistent with a hybrid ‘superswarm’. This massive hybridization and population formation event(s) occurred over approximately 140 years and likely fuelled the successful invasion of a diverse range of habitats. The implications are that multiple colonizations coupled with hybridization can lead to the formation of new stable genetic populations potentially kick-starting speciation and adaptive radiation over a very short time.

Diagnosis of diabetes insipidus observed in Swiss Duroc boars

Background Diabetes insipidus (DI) is a rare disease in humans and animals, which is caused by the lack of production, malfunction or dysfunction of the distal nephron to the antidiuretic effect of the antidiuretic hormone (ADH). Diagnosis requires a thorough medical history, clinical examination and further laboratory confirmation. This case report describes the appearance of DI in five Duroc boars in Switzerland. Case presentation Two purebred intact Duroc boars at the age of 8 months and 1.5 years, respectively, with a history of polyuric and polydipsic symptoms had been referred to the Swine Clinic in Berne. Based on the case history, the results of clinical examination and the analysis of blood and urine, a tentative diagnosis of DI was concluded. Finally, the diagnosis was confirmed by findings from a modified water deprivation test, macroscopic examinations and histopathology. Following the diagnosis, three genes known to be involved in inherited DI in humans were analyzed in order to explore a possible genetic background of the affected boars. Conclusion The etiology of DI in pigs is supposed to be the same as in humans, although this disease has never been described in pigs before. Thus, although occurring only on rare occasions, DI should be considered as a differential diagnosis in pigs with polyuria and polydipsia. It seems that a modified water deprivation test may be a helpful tool for confirming a diagnosis in pigs. Since hereditary forms of DI have been described in humans, the occurrence of DI in pigs should be considered in breeding programs although we were not able to identify a disease associated mutation.

Variable phenotypic expressivity in inbred retinal degeneration mouse lines: A comparative study of C3H/HeOu and FVB/N rd1 mice

PURPOSE Recent advances in optogenetics and gene therapy have led to promising new treatment strategies for blindness caused by retinal photoreceptor loss. Preclinical studies often rely on the retinal degeneration 1 (rd1 or Pde6b(rd1)) retinitis pigmentosa (RP) mouse model. The rd1 founder mutation is present in more than 100 actively used mouse lines. Since secondary genetic traits are well-known to modify the phenotypic progression of photoreceptor degeneration in animal models and human patients with RP, negligence of the genetic background in the rd1 mouse model is unwarranted. Moreover, the success of various potential therapies, including optogenetic gene therapy and prosthetic implants, depends on the progress of retinal degeneration, which might differ between rd1 mice. To examine the prospect of phenotypic expressivity in the rd1 mouse model, we compared the progress of retinal degeneration in two common rd1 lines, C3H/HeOu and FVB/N. METHODS We followed retinal degeneration over 24 weeks in FVB/N, C3H/HeOu, and congenic Pde6b(+) seeing mouse lines, using a range of experimental techniques including extracellular recordings from retinal ganglion cells, PCR quantification of cone opsin and Pde6b transcripts, in vivo flash electroretinogram (ERG), and behavioral optokinetic reflex (OKR) recordings. RESULTS We demonstrated a substantial difference in the speed of retinal degeneration and accompanying loss of visual function between the two rd1 lines. Photoreceptor degeneration and loss of vision were faster with an earlier onset in the FVB/N mice compared to C3H/HeOu mice, whereas the performance of the Pde6b(+) mice did not differ significantly in any of the tests. By postnatal week 4, the FVB/N mice expressed significantly less cone opsin and Pde6b mRNA and had neither ERG nor OKR responses. At 12 weeks of age, the retinal ganglion cells of the FVB/N mice had lost all light responses. In contrast, 4-week-old C3H/HeOu mice still had ERG and OKR responses, and we still recorded light responses from C3H/HeOu retinal ganglion cells until the age of 24 weeks. These results show that genetic background plays an important role in the rd1 mouse pathology. CONCLUSIONS Analogous to human RP, the mouse genetic background strongly influences the rd1 phenotype. Thus, different rd1 mouse lines may follow different timelines of retinal degeneration, making exact knowledge of genetic background imperative in all studies that use rd1 models.

Update on food allergy

Food allergies are a global health issue with increasing prevalence. Allergic reactions can range from mild local symptoms to severe anaphylactic reactions. Significant progress has been made in diagnostic tools such as component-resolved diagnostics and its impact on risk stratification as well as in therapeutic approaches including biologicals. However, a cure for food allergy has not yet been achieved and patients and their families are forced to alter eating habits and social engagements, impacting their quality of life. New technologies and improved in vitro and in vivo models will advance our knowledge of the pathogenesis of food allergies and multicenter-multinational cohort studies will elucidate interactions between genetic background, lifestyle, and environmental factors. This review focuses on new insights and developments in the field of food allergy and summarizes recently published articles.

Streptococcus pneumoniae capsule determines disease severity in experimental pneumococcal meningitis.

Streptococcus pneumoniaebacteria can be characterized into over 90 serotypes according to the composition of their polysaccharide capsules. Some serotypes are common in nasopharyngeal carriage whereas others are associated with invasive disease, but when carriage serotypes do invade disease is often particularly severe. It is unknown whether disease severity is due directly to the capsule type or to other virulence factors. Here, we used a clinical pneumococcal isolate and its capsule-switch mutants to determine the effect of capsule, in isolation from the genetic background, on severity of meningitis in an infant rat model. We found that possession of a capsule was essential for causing meningitis. Serotype 6B caused significantly more mortality than 7F and this correlated with increased capsule thickness in the cerebrospinal fluid (CSF), a stronger inflammatory cytokine response in the CSF and ultimately more cortical brain damage. We conclude that capsule type has a direct effect on meningitis severity. This is an important consideration in the current era of vaccination targeting a subset of capsule types that causes serotype replacement.

Esx1 dosage impacts reproductive fitness: Effects on viability and fertility

Numerous genes expressed in placenta or testis localize to the X-chromosome. Both tissues undergo specialized X-chromosome inactivation (imprinted paternal inactivation in placenta and MSCI in testicular germ cells). When the X-chromosome is duplicated or improperly inactivated, defects in placentation, growth and spermatogenesis are noted, suggesting tight control of X-chromosome gene dosage is important for reproduction. ^ Esx1 is a mouse homeobox gene on the X-chromosome with expression limited to extraembryonic tissues and testicular germ cells. Here, we examine the effects of increased and decreased Esx1 dosage on placental and testicular development, the role of genetic background on Esx1 function and characterize the human orthologue of Esx1. ^ Previously, by targeted deletion, Esx1 was shown to be an X-chromosome imprinted regulator of placental development and fetal growth. We show C57Bl6-congenic Esx1 mutants display a more severe phenotype with decreased viability and that the 129 genetic background contains dominant modifier genes that enhance Esx1 mutant survival. ^ Varying Esx1 dosage impacts testicular germ cell development. Esx1 hemizygous null mice are fertile, but we show their testes are two-thirds normal size. To examine the effect of increased Esx1 dosage, Esx1 BAC transgenic mice were generated. Increased Esx1 dosage results in dramatic deficits in testicular germ cell development, leading to sterility and testes one-fourth normal size. We show germ cell loss occurs through apoptosis, begins between postnatal day 6 and 10, and that no spermatocytes complete meiosis. Interestingly, increased Esx1 dosage in testes mimics germ cell loss seen in Klinefelter's (XXY) mice and humans and may represent a molecular mechanism for the infertility characteristic of this syndrome. ^ Esx1 dosage impacts reproductive fitness when maternally transmitted. Three transgenic founder females were unable to transmit the transgene to live offspring, but did produce transgenic pups at earlier stages. Additionally, one line of Esx1 BAC transgenic mice demonstrated decreased embryo size and fitness when the transgene is inherited compared to wild type littermates. ^ It is possible that Esx1 plays a role in human disorders of pregnancy, growth and spermatogenesis. Therefore, we cloned and characterized ESX1L (human Esx1), and show it is expressed in human testis and placenta. ^

Requirement of GCN5 histone acetyltransferase in mouse neural tube closure and skeletal patterning

Histone acetylation plays an essential role in many DNA-related processes such as transcriptional regulation via modulation of chromatin structure. Many histone acetytransferases have been discovered and studied in the past few years, but the roles of different histone acetyltransferases (HAT) during mammalian development are not well defined at present. Gcn5 histone acetyltransferase is highly expressed until E16.5 during development. Previous studies in our lab using a constitutive null allele demonstrated that Gcn5 knock out mice are embryonic lethal, precluding the study of Gcn5 functions at later developmental stages. The creation of a conditional Gcn5 null allele, Gcn5flox allele, bypasses the early lethality. Mice homozygous for this allele are viable and appear healthy. In contrast, mice homozygous for a Gcn5 Δex3-18 allele created by Cre-loxP mediated deletion display a phenotype identical to our original Gcn5 null mice. Strikingly, a Gcn5flox(neo) allele, which contain a neomycin cassette in the second intron of Gcn5 is only partially functional and gives rise to a hypomorphic phenotype. Initiation of cranial neural tube closure at forebrain/midbrain boundary fails, resulting in an exencephaly in some Gcn5flox(neo)/flox(neo) embryos. These defects were found at an even greater penetrance in Gcn5flox(neo)/Δ embryos and become completely penetrant in the 129Sv genetic background, suggesting that Gcn5 controls mouse neural tube closure in a dose dependent manner. Furthermore, both Gcn5flox(neo)/flox(neo) and Gcn5 flox(neo)/Δ embryos exhibit anterior homeotic transformations in lower thoracic and lumbar vertebrae. These defects are accompanied by decreased expression levels and a shift in anterior expression boundary of Hoxc8 and Hoxc9. This study provides the first evidence that Gcn5 regulates Hox gene expression and is required for normal axial skeletal patterning in mice. ^

Acute survivorship of the deep-sea coral Lophelia pertusa from the Gulf of Mexico under acidification,warming,and deoxygenation

Changing global climate due to anthropogenic emissions of CO2 are driving rapid changes in the physical and chemical environment of the oceans via warming, deoxygenation, and acidification. These changes may threaten the persistence of species and populations across a range of latitudes and depths, including species that support diverse biological communities that in turn provide ecological stability and support commercial interests. Worldwide, but particularly in the North Atlantic and deep Gulf of Mexico, Lophelia pertusa forms expansive reefs that support biological communities whose diversity rivals that of tropical coral reefs. In this study, L. pertusa colonies were collected from the Viosca Knoll region in the Gulf of Mexico (390 to 450 m depth), genotyped using microsatellite markers, and exposed to a series of treatments testing survivorship responses to acidification, warming, and deoxygenation. All coral nubbins survived the acidification scenarios tested, between pH of 7.67 and 7.90 and aragonite saturation states of 0.92 and 1.47. However, calcification generally declined with respect to pH, though a disparate response was evident where select individuals net calcified and others exhibited net dissolution near a saturation state of 1. Warming and deoxygenation both had negative effects on survivorship, with up to 100% mortality observed at temperatures above 14ºC and oxygen concentrations of approximately 1.5 ml·l-1. These results suggest that, over the short-term, climate change and OA may negatively impact L. pertusa in the Gulf of Mexico, though the potential for acclimation and the effects of genetic background should be considered in future research.

DNA methylation and adaptive response in forest tree species

Progressive increase of temperatures as well as longer seasonal drought periods revealed by climate studies correspond to fast environmental changes that forest species face with their actual genetic background. Natural selective processes cannot develop an adaptive response within this time frame. Thus the capability of forest tree species to adapt to the new environments will depend on their genetic background, but also rely on their phenotypic plasticity. Several reports have shown the involvement of epigenetic modifiers as the basis of the phenotypic plasticity, and in particular to the adaptation to abiotic stresses. DNA methylation (methylation of cytosine residues)is one the most important epigenetic modification in eukaryotes. Itis involved in specific biological processes such as gene transcription regulation, gene silencing, mobile element control or genome imprinting.Therefore, there is a great interest in analyzing cytosine methylation levels and distribution within the genome

Removing exogenous information using pedigree data

Management of certain populations requires the preservation of its pure genetic background. When, for different reasons, undesired alleles are introduced, the original genetic conformation must be recovered. The present study tested, through computer simulations, the power of recovery (the ability for removing the foreign information) from genealogical data. Simulated scenarios comprised different numbers of exogenous individuals taking partofthe founder population anddifferent numbers of unmanaged generations before the removal program started. Strategies were based on variables arising from classical pedigree analyses such as founders? contribution and partial coancestry. The ef?ciency of the different strategies was measured as the proportion of native genetic information remaining in the population. Consequences on the inbreeding and coancestry levels of the population were also evaluated. Minimisation of the exogenous founders? contributions was the most powerful method, removing the largest amount of genetic information in just one generation.However, as a side effect, it led to the highest values of inbreeding. Scenarios with a large amount of initial exogenous alleles (i.e. high percentage of non native founders), or many generations of mixing became very dif?cult to recover, pointing out the importance of being careful about introgression events in population

Influencia de la castración y la línea genética paterna sobre el rendimiento productivo y la calidad de la canal y de la carne en cerdos sacrificados a pesos elevados

El objetivo general de esta Tesis Doctoral fue estudiar la influencia del sexo, el método de castración de los machos y la línea genética paterna sobre la productividad y la calidad de la canal y de la carne en cerdos blancos sacrificados a pesos elevados con destino a la industria de los productos curados de calidad. En el experimento 1, se utilizaron 360 cerdos sacrificados a 125 kg de peso vivo (PV) para estudiar la influencia del sexo y la castración [machos inmunocastrados (MI), machos castrados quirúrgicamente (MC) y hembras enteras (HE)] de dos líneas genéticas paternas Large White (Top York y Tempo) sobre los rendimientos productivos y la calidad de la canal y de la carne. La línea materna utilizada fue Large White × Landrace en todos los casos. Los MI se inmunizaron contra el factor de liberación de gonadotropina (GnRF) mediante la utilización de Improvac a los 78 (16 d en prueba) y 126 (64 d en prueba y 48 d antes del sacrificio) d de edad. Cada uno de los 6 tratamientos experimentales fue replicado 6 veces (cuadra con 10 cerdos). Desde el inicio de la prueba hasta el día de la primera inyección con Improvac (62 a 78 d de edad) los MI y las HE crecieron menos (P < 0,001) que los MC sin que se observaran diferencias en el consumo medio diario de pienso (CMD). Los MC tuvieron peor eficiencia alimenticia que las HE con los MI mostrando valores intermedios (P < 0,01). Entre las dos inyecciones de Improvac (78 a 126 d de edad), los MI crecieron y comieron menos que los MC, mostrando las HE valores intermedios (P < 0,001). Los MI fueron más eficientes que las HE y ambos más eficientes que los MC (P < 0,001). Sin embargo, desde la segunda inyección de Improvac hasta el sacrificio (126 a 174 d de edad) los MI crecieron más y fueron más eficientes (P < 0,001) que las HE y los MC. Al final de la prueba, MI y MC crecieron más (P < 0,01) que HE. Asimismo, los MI fueron más eficientes (P < 0,001) pero presentaron menor rendimiento de canal (P < 0,001) que los MC y las HE. Por otro lado, los MI y las HE depositaron menos grasa dorsal que los MC (P < 0,001). Las hembras tuvieron mayor rendimiento de lomo y menos grasa intramuscular que MI y MC (P < 0,01). Asimismo, las HE tuvieron mayor rendimiento de jamón en fresco y perfilado que los MC con los MI mostrando valores intermedios (P < 0,05). Los cerdos híbridos procedentes de machos Tempo crecieron más (P < 0,001) que los procedentes de machos Top York, sin que se encontraran diferencias para el CMD o para la eficiencia alimenticia. Los híbridos de los cruces con Top York tuvieron mejores rendimientos de jamones frescos y perfilados (P < 0,05) pero menor rendimiento de lomo y menos grasa intramuscular que los cruces con Tempo (P < 0,01). En conclusión, los MI presentaron mejor eficiencia alimenticia, pero menor rendimiento de canal que los MC y las HE. El contenido en grasa intramuscular fue similar entre MC y MI y superior para ambos que para las HE. Los cruces procedentes de la línea paterna Tempo crecieron más y tuvieron mayor contenido en grasa intramuscular, pero un rendimiento en jamón perfilado ligeramente inferior al de los cruces procedentes de la línea paterna Top York. Se concluye que la inmunocastración de los machos es una alternativa viable a la castración quirúrgica para la producción de cerdos pesados destinados a la industria de los productos curados. Debido a su mayor potencial de crecimiento y mayor contenido en grasa intramuscular, los híbridos procedentes de la línea paterna Tempo presentan ventajas frente a los híbridos procedentes de la línea paterna Top York cuando se destinan a la industria de productos curados de calidad. En el experimento 2, se utilizaron 240 cerdos para comparar los rendimientos productivos y los parámetros de calidad de la canal de MI, MC y HE destinados a la industria de productos cárnicos curados procedentes del cruce de la línea materna Large White × Landrace con la línea genética paterna Duroc o Pietrain. Entre las 2 inyecciones de Improvac (87 a 137 d de edad), los MI y las HE crecieron menos que los MC (P < 0,01). Asimismo, los MI comieron menos pienso que las HE y ambos menos que los MC (2,33, 2,55 y 2,77 kg/d; respectivamente; P < 0,001). Como resultado, los MI fueron más eficientes que los MC y las HE (P < 0,001). Desde la segunda inyección de Improvac hasta el momento del sacrificio (137 a 164 d de edad), los MI fueron más eficientes que las HE y ambos más que los MC (0,346, 0,323 y 0,300, respectivamente; P < 0,001). Las diferencias observadas en este periodo entre los sexos en cuanto a rendimientos productivos fueron más pronunciadas en los cerdos procedentes de la línea paterna Pietrain que los de la línea Duroc (P < 0,05 para la interacción). En el global de la prueba (87 a 164 d de edad) el sexo no afectó al crecimiento en los cerdos procedentes de la línea paterna Duroc pero en los cerdos procedentes de la línea paterna Pietrain, los MI y los MC crecieron más que las HE (P < 0,05 para la interacción). Asimismo, los MI tuvieron mejor eficiencia alimenticia (0,406, 0,364 y 0,380, P < 0,001) y menor rendimiento de la canal (76,6, 78,1 y 78,8%; P < 0,001) que los MC y las HE. Las canales de las HE fueron más magras que las canales de los MC, con las canales de los MI mostrando valores intermedios (P < 0,01). El rendimiento en jamones y lomos fue mayor para las HE que para los MI y los MC (P < 0,001). El contenido en grasa intramuscular fue menor en las HE que en los MC, con los MI mostrando valores intermedios (3,5 vs. 3,9 y 3,7%; P < 0,05). Por otra parte, los híbridos procedentes de machos Duroc crecieron más rápido (1,167 vs. 0,986 kg/d; P < 0,001), consumieron más pienso (3,07 vs. 2,56 kg/d; P < 0,001) y tuvieron más grasa intramuscular (P < 0,001), pero menor rendimiento en jamones y lomos (P < 0,01) que los híbridos procedentes de machos Pietrain. Se concluye que los MI presentaron mejores productividades pero menores rendimientos de canal que MC y HE. El contenido en grasa intramuscular en el músculo longissimus dorsi fue menor para las HE que para los MC con valores intermedios para los MI. Los cruces procedentes de la genética paterna Duroc crecieron más y tuvieron más grasa intramuscular pero menos rendimiento de jamón que los cerdos procedentes de machos Pietrain. Por tanto, los MI deben ser preferidos a los MC y los cruces con la línea paterna Duroc deben ser preferidos a los cruces con Pietrain para producir canales cuando sus partes nobles están destinadas a la industria de productos cárnicos curados. En base a estos resultados, se concluye que la inmunocastración es una alternativa factible a la castración quirúrgica y que líneas genéticas paternas Tempo y Duroc son mejores para la producción de cerdo blanco pesado que las líneas Top York y Pietrain. Las interacciones entre el sexo y las líneas genéticas paternas estudiadas, sugieren que el resultado final depende en parte de la línea genética paterna utilizada. En cualquier caso, la inmunocastración es una alternativa factible a la castración quirúrgica para la producción de canales destinadas a la industria de los productos cárnicos curados. ABSTRACT The general aim of this PhD Thesis was to study the influence of sex, method of castration, and genetic background of the sire line on growth performance and carcass and meat quality merits of heavy white pigs destined to the dry-cured industry. In experiment 1, 360 pigs slaughtered at 125 kg of body weight were used to study the influence of sex and castration methodology [immunocastrated males (ICM), surgically castrated males (SCM), and intact females (IF)] of 2 terminal Large White sire lines (Top York and Tempo) on growth performance and carcass and meat quality. The female line was Large White × Landrace in all cases. The ICM pigs were immunized against gonadotropin-releasing factor with Improvac at 78 (16 d on trial) and 126 (64 d on trial and 48 d before slaughter) d of age. Each of the 6 treatments was replicated 6 times (10 pigs/pen). From the start of the experiment to the day of the first Improvac injection (62 to 78 d of age), ICM and IF grew slowlier (P < 0.001) than SCM but no differences in feed intake were detected. The SCM pigs had greater gain to feed ratio (G:F) than the IF with the ICM pigs being intermediate (P < 0.01). Between the 2 Improvac injections (78 to 126 d of age), the ICM pigs ate less feed (P < 0.001) and grew slowlier rate than the SCM pigs, with the growth of IF being intermediate. The ICM pigs were more efficient than the IF, and both were more efficient than the SCM pigs (P < 0.001). However, from the second Improvac injection to slaughter (126 to 174 d of age), the ICM pigs grew at a faster rate (P < 0.001) and were more efficient (P < 0.001) than the IF and the SCM pigs. Cumulatively, ICM and SCM pigs grew faster (P < 0.01) than IF and the ICM pigs were more efficient than the other two sexes (P < 0.001). However, the ICM pigs had reduced (P < 0.001) carcass yield compared with SCM and IF. The ICM and IF pigs also had less (P < 0.001) backfat depth than the SCM pigs. Intact females had higher (P < 0.01) loin yield but less intramuscular fat (P < 0.01) than ICM and SCM pigs and higher (P < 0.05) fresh and trimmed ham yields than SCM pigs, with ICM pigs being intermediate. Crossbreds from the Tempo sires grew faster (P < 0.001) than crossbreds from the Top York sires but no differences (P > 0.10) were detected for feed intake or feed efficiency. Crossbreds from the Top York sires had higher (P < 0.05) fresh and trimmed ham yields but less (P < 0.01) loin yield and intramuscular fat content than crossbreds from the Tempo sires. In conclusion, ICM pigs are more efficient, but have less carcass yield than SCM and IF pigs. The intramuscular fat content was lowest for the IF and similar for ICM and SCM pigs. Crossbreds from Tempo sires were heavier and had greater intramuscular fat content, but had less trimmed ham yield as compared with crossbreds from the Top York sires. Immunocastrated pigs can replace SCM pigs for the production of heavy pigs destined for the dry-cured industry. Because of increased carcass weight and the higher intramuscular content, crossbreds from Tempo sires may have an advantage over crossbreds from Top York sires for the dry-cured industry. In experiment 2, a total of 240 pigs were used to compare growth performance and carcass quality traits of immunocastrated males, surgically castrated males, and intact females of crossbreds from Large White × Landrace females and Duroc or Pietrain sires destined to the dry-cured industry. Between the 2 Improvac injections (87 and 137 d of age), ICM and IF pigs had lower average daily gain (ADG) than SCM pigs (P < 0.01). Also, ICM pigs ate less feed than IF and both type of pigs ate less than SCM pigs (2.33, 2.55, and 2.77 kg/d; P < 0.001). Consequently, ICM pigs had better G:F than SCM and IF (P < 0.001). From the second Improvac injection to slaughter (137 to 164 d of age), ICM pigs were more efficient than IF and both were more efficient than SCM pigs (0.346, 0.323, and 0.300 g/g; P < 0.001). The differences in growth performance among genders observed in this period were more pronounced for the Pietrain than for the Duroc crossbreds (P < 0.05 for the interaction). For the entire experimental period (87 to 164 d of age), gender did not affect ADG for Duroc crossbreds but for Pietrain crossbreds ICM and SCM had higher ADG than IF (P < 0.05 for the interaction). The ICM pigs had better feed efficiency (0.406, 0.364, and 0.380; g/g; P < 0.001) and lower carcass yield (76.6, 78.1, and 78.8%; P < 0.001) than SCM or IF. Carcasses from IF were leaner than carcasses from SCM with carcasses from ICM being intermediate (P < 0.01). Ham and loin (P < 0.001) yields were higher for IF than for ICM or SCM pigs. Intramuscular fat content was lower for IF than for SCM pigs with that of ICM pigs being intermediate (3.5 vs. 3.9 and 3.7%; P < 0.05). Cumulatively, crossbreds from Duroc sires had higher ADG (1.167 vs. 0.986 kg/d; P < 0.001) and average daily feed intake (3.07 vs. 2.56 kg/d; P < 0.001) and more intramuscular fat (P < 0.001) but less ham and loin yields (P < 0.01) than crossbreds from Pietrain sires. It is concluded that growth performance was better, but carcass yield lower, for ICM pigs than for SCM and IF. Intramuscular fat content in longissimus dorsi muscle was lower for IF than for SCM pigs with ICM pigs being intermediate. Crossbreds from Duroc sires grew faster and had more intramuscular fat but less ham yield than crossbreds from Pietrain sires. Therefore, ICM pigs should be preferred to SCM pigs, and Duroc crossbreds should be preferred to Pietrain crossbreds to produce carcasses destined to the production of primal cuts for the dry-cured industry. We conclude that immunocastration might be a sound alternative to surgical castration in pigs and that Tempo and Duroc might be better for the production of heavy pigs than Top York and Pietrain. The interactions reported between sex and genetic sire line, suggested that the benefits of immunocastration as an alternative to surgical castration might depend at least part on the sire line used. In any case, immunocastration is a good alternative to surgical castration for the production of carcasses destined to the dry-cured industry.

Abnormal skeletal patterning in embryos lacking a single Cbp allele: A partial similarity with Rubinstein–Taybi syndrome

CBP is a transcriptional coactivator required by many transcription factors for transactivation. Rubinstein–Taybi syndrome, which is an autosomal dominant syndrome characterized by abnormal pattern formation, has been shown to be associated with mutations in the Cbp gene. Furthermore, Drosophila CBP is required in hedgehog signaling for the expression of decapentapleigic, the Drosophila homologue of bone morphogenetic protein. However, no direct evidence exists to indicate that loss of one copy of the mammalian Cbp gene affects pattern formation. Here, we show that various abnormalities occur at high frequency in the skeletal system of heterozygous Cbp-deficient mice resulting from a C57BL/6-CBA × BALB/c cross. In support of a conserved signaling pathway for pattern formation in insects and mammals, the expression of Bmp7 was found to be reduced in the heterozygous mutants. The frequency of the different abnormalities was significantly lower in a C57BL/6-CBA background, suggesting that the genetic background is an important determinant of the variability and severity of the anomalies seen in Rubinstein–Taybi syndrome patients.