Regulation of GABA(A) and glutamate receptor expression, synaptic facilitation and long-term potentiation in the hippocampus of prion mutant mice

Background: Prionopathies are characterized by spongiform brain degeneration, myoclonia, dementia, and periodic electroencephalographic (EEG) disturbances. The hallmark of prioniopathies is the presence of an abnormal conformational isoform (PrP(sc)) of the natural cellular prion protein (PrP(c)) encoded by the Prnp gene. Although several roles have been attributed to PrP(c), its putative functions in neuronal excitability are unknown. Although early studies of the behavior of Prnp knockout mice described minor changes, later studies report altered behavior. To date, most functional PrP(c) studies on synaptic plasticity have been performed in vitro. To our knowledge, only one electrophysiological study has been performed in vivo in anesthetized mice, by Curtis and coworkers. They reported no significant differences in paired-pulse facilitation or LTP in the CA1 region after Schaffer collateral/commissural pathway stimulation. Principal Findings: Here we explore the role of PrP(c) expression in neurotransmission and neural excitability using wild-type, Prnp -/- and PrP(c)-overexpressing mice (Tg20 strain). By correlating histopathology with electrophysiology in living behaving mice, we demonstrate that both Prnp -/- mice but, more relevantly Tg20 mice show increased susceptibility to KA, leading to significant cell death in the hippocampus. This finding correlates with enhanced synaptic facilitation in paired-pulse experiments and hippocampal LTP in living behaving mutant mice. Gene expression profiling using Illumina microarrays and Ingenuity pathways analysis showed that 129 genes involved in canonical pathways such as Ubiquitination or Neurotransmission were co-regulated in Prnp -/- and Tg20 mice. Lastly, RT-qPCR of neurotransmission-related genes indicated that subunits of GABA(A) and AMPA-kainate receptors are co-regulated in both Prnp -/- and Tg20 mice. Conclusions/Significance: Present results demonstrate that PrP(c) is necessary for the proper homeostatic functioning of hippocampal circuits, because of its relationships with GABA(A) and AMPA-Kainate neurotransmission. New PrP(c) functions have recently been described, which point to PrP(c) as a target for putative therapies in Alzheimer's disease. However, our results indicate that a "gain of function" strategy in Alzheimer's disease, or a "loss of function" in prionopathies, may impair PrP(c) function, with devastating effects. In conclusion, we believe that present data should be taken into account in the development of future therapies.

Surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry: A tool to predict pork quality

Expression of water soluble proteins of fresh pork Longissimus thoracis from 4 pure breed pigs (Duroc, Large White, Landrace, and Piétrain) was studied to identify candidate protein markers for meat quality. Surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS) was used to obtain the soluble protein profiles of Longissimus thoracis muscles. The pure breeds showed differences among the studied meat quality traits (pHu, drip loss, androstenone, marbling, intramuscular fat, texture, and moisture), but no significant differences were detected in sensory analysis. Associations between protein peaks obtained with SELDI-TOF-MS and meat quality traits, mainly water holding capacity, texture and skatole were observed. Of these peaks, a total of 10 peaks from CM10 array and 6 peaks from Q10 array were candidate soluble protein markers for pork loin quality. The developed models explained a limited proportion of the variability, however they point out interesting relationships between protein expression and meat quality

Analysis of raw hams using SELDI-TOF-MS to predict the final quality of dry-cured hams

The relationship between protein profiles of Gluteus medius (GM) muscles of raw hams obtained from 4 pure breed pigs (Duroc, Large White, Landrace, and Piétrain) with the final quality of the Semimembranosus and Biceps femoris muscles of dry-cured hams was investigated. As expected, Duroc hams showed higher levels of marbling and intramuscular fat content than the other breeds. Piétrain hams were the leanest and most conformed, and presented the lowest salt content in dry-cured hams. Even if differences in the quality traits (colour, water activity, texture, composition, intramuscular fat, and marbling) of dry-cured hams were observed among the studied breeds, only small differences in the sensory attributes were detected. Surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS) was used to obtain the soluble protein profiles of GM muscles. Some associations between protein peaks obtained with SELDI-TOF-MS and quality traits, mainly colour (b*) and texture (F0, Y2, Y90) were observed. Candidate protein markers for the quality of processed dry-cured hams were identified

Malignant pheochromocytoma in a pig

Abstract. Endocrine tumors are rarely observed in pigs, and pheochromocytomas have been only punctually described. The current report describes a white and firm, 15-cm in diameter, neoplastic mass located in the adrenal gland with metastasis to regional lymph nodes in a 2.5-year-old sow. The masses had marked desmoplasia that supported a population of polygonal-tospindle– shaped neoplastic cells arranged into cords and packets within a delicate fibrovascular stroma. Immunohistochemical staining of the tumor was positive for chromogranin and negative for neurofilament protein in adrenal and lymph node masses, which was characteristic of a malignant pheochromocytoma.

Minimal requirements for parameters to be used for the development of predictive models for microbial source tracking. The example of the pair somatic coliphages and phages infecting Bacteroides

Several methods and approaches for measuring parameters to determine fecal sources of pollution in water have been developed in recent years. No single microbial or chemical parameter has proved sufficient to determine the source of fecal pollution. Combinations of parameters involving at least one discriminating indicator and one universal fecal indicator offer the most promising solutions for qualitative and quantitative analyses. The universal (nondiscriminating) fecal indicator provides quantitative information regarding the fecal load. The discriminating indicator contributes to the identification of a specific source. The relative values of the parameters derived from both kinds of indicators could provide information regarding the contribution to the total fecal load from each origin. It is also essential that both parameters characteristically persist in the environment for similar periods. Numerical analysis, such as inductive learning methods, could be used to select the most suitable and the lowest number of parameters to develop predictive models. These combinations of parameters provide information on factors affecting the models, such as dilution, specific types of animal source, persistence of microbial tracers, and complex mixtures from different sources. The combined use of the enumeration of somatic coliphages and the enumeration of Bacteroides-phages using different host specific strains (one from humans and another from pigs), both selected using the suggested approach, provides a feasible model for quantitative and qualitative analyses of fecal source identification.

The effect of age at the first mating and herd size on the lifetime productivity of sows

The effect of age at the first mating and herd size were evaluated in the reference Spanish Databank (BDporc) of 37 698 sows born between 1991 and 1995 and with individual lifetime records. The data included dates of births at entrance and culling, first mating, repetitive mating and conception, first farrowing and weaning records. Individual records were validated before the analysis by screening them through a tolerance “filter” in order to eliminate the extreme values from the analysis. The total database of the sows was classified in 7 classes according to age at the first mating (< 210, 210–220, 221–230, 231–240, 241–250, 251–270, and > 270 days) and in 6 classes of herd size (< 200, 200–300, 301–400, 401–600, 601–800, and > 800 sows). The total number of litters and number of weaned piglets obtained from each sow during the lifetime production were significantly (P < 0.05) greater for gilts between 221 and 240 d of age at the first mating. There was a significant (P < 0.001) effect of the herd size on the reproductive performance of the sow, and the best performance was obtained with herds with 401 to 600 sows compared to < 200 or > 800 sow-herds. Furthermore, a significant (P < 0.001) interaction between age at the first mating and herd size was detected and can be associated with a particular pattern for the herd size class 401–600 sows with the best performances obtained for the sows first mated at less than 200 days. For the other herd sizes, the results indicated that sows mated for the first time at the right age, 221–240 days, are more productive, both in the number and size of the parities throughout lifetime production.

Copy number variation in the porcine genome inferred from a 60 k SNP BeadChip

Background: Recent studies in pigs have detected copy number variants (CNVs) using the Comparative Genomic Hybridization technique in arrays designed to cover specific porcine chromosomes. The goal of this study was to identify CNV regions (CNVRs) in swine species based on whole genome SNP genotyping chips. Results: We used predictions from three different programs (cnvPartition, PennCNV and GADA) to analyze data from the Porcine SNP60 BeadChip. A total of 49 CNVRs were identified in 55 animals from an Iberian x Landrace cross (IBMAP) according to three criteria: detected in at least two animals, contained three or more consecutive SNPs and recalled by at least two programs. Mendelian inheritance of CNVRs was confirmed in animals belonging to several generations of the IBMAP cross. Subsequently, a segregation analysis of these CNVRs was performed in 372 additional animals from the IBMAP cross and its distribution was studied in 133 unrelated pig samples from different geographical origins. Five out of seven analyzed CNVRs were validated by real time quantitative PCR, some of which coincide with well known examples of CNVs conserved across mammalian species. Conclusions: Our results illustrate the usefulness of Porcine SNP60 BeadChip to detect CNVRs and show that structural variants can not be neglected when studying the genetic variability in this species.

Applying precision feeding techniques in growing-finishing pig operations

The high cost of feed ingredients, the use of non-renewable sources of phosphate and the dramatic increase in the environmental load resulting from the excessive land application of manure are major challenges for the livestock industry. Precision feeding is proposed as an essential approach to improve the utilization of dietary nitrogen, phosphorus and other nutrients and thus reduce feeding costs and nutrient excretion. Precision feeding requires accurate knowledge of the nutritional value of feedstuffs and animal nutrient requirements, the formulation of diets in accordance with environmental constraints, and the gradual adjustment of the dietary nutrient supply to match the requirements of the animals. After the nutritional potential of feed ingredients has been precisely determined and has been improved by the addition of enzymes (e.g. phytases) or feed treatments, the addition of environmental objectives to the traditional feed formulation algorithms can promote the sustainability of the swine industry by reducing nutrient excretion in swine operations with small increases in feeding costs. Increasing the number of feeding phases can also contribute to significant reductions in nutrient excretion and feeding costs. However, the use of precision feeding techniques in which pigs are fed individually with daily tailored diets can further improve the efficiency with which pigs utilize dietary nutrients. Precision feeding involves the use of feeding techniques that allow the provision of the right amount of feed with the right composition at the right time to each pig in the herd. Using this approach, it has been estimated that feeding costs can be reduced by more than 4.6%, and nitrogen and phosphorus excretion can both be reduced by more than 38%. Moreover, the integration of precision feeding techniques into large-group production systems can provide real-time off-farm monitoring of feed and animals for optimal slaughter and production strategies, thus improving the environmental sustainability of pork production, animal well-being and meat-product quality.

Selection of internal control genes for real-time quantitative PCR in ovary and uterus of sows across pregnancy

Reproductive traits play a key role in pig production in order to reduce costs and increase economic returns. Among others, gene expression analyses represent a useful approach to study genetic mechanisms underlying reproductive traits in pigs. The application of reverse-transcription quantitative PCR requires the selection of appropriate reference genes, whose expression levels should not be affected by the experimental conditions, especially when comparing gene expression across different physiological stages.

Minimal requirements for parameters to be used for the development of predictive models for microbial source tracking. The example of the pair somatic coliphages and phages infecting Bacteroides

Several methods and approaches for measuring parameters to determine fecal sources of pollution in water have been developed in recent years. No single microbial or chemical parameter has proved sufficient to determine the source of fecal pollution. Combinations of parameters involving at least one discriminating indicator and one universal fecal indicator offer the most promising solutions for qualitative and quantitative analyses. The universal (nondiscriminating) fecal indicator provides quantitative information regarding the fecal load. The discriminating indicator contributes to the identification of a specific source. The relative values of the parameters derived from both kinds of indicators could provide information regarding the contribution to the total fecal load from each origin. It is also essential that both parameters characteristically persist in the environment for similar periods. Numerical analysis, such as inductive learning methods, could be used to select the most suitable and the lowest number of parameters to develop predictive models. These combinations of parameters provide information on factors affecting the models, such as dilution, specific types of animal source, persistence of microbial tracers, and complex mixtures from different sources. The combined use of the enumeration of somatic coliphages and the enumeration of Bacteroides-phages using different host specific strains (one from humans and another from pigs), both selected using the suggested approach, provides a feasible model for quantitative and qualitative analyses of fecal source identification.

Estudi tècnic-econòmic de l'adaptació a la Llei de Benestar Animal d'una explotació porcina de la comarca d'Osona i posterior valoració de la seva implementació

Des de la publicació de la Llei de Benestar Animal RD 1135/ 2002, de 31 d’octubre que regula els tres aspectes clau de la producció porcina‐ sistemes d’estabulació i construccions permeses; formació obligatòria dels ramaders i maneig dels animals‐ totes les granges de nova construcció han estat projectades seguint els seus preceptes. Mentre que les granges existents han hagut d’adaptar‐hi les seves instal∙lacions sota amenaça de tancament si no es complien els requisits. L’objectiu d’aquest treball ha estat l’elaboració de l’estudi tècnic‐ econòmic per a l’adaptació a la Llei de Benestar Animal d’una explotació porcina de la comarca d’Osona. Després d’avaluar la situació inicial de les instal∙lacions, establir les línies de previsió de creixement i analitzar els avantatges i inconvenients de cada sistema i model, s’opta pel sistema d’alimentació electrònic model Nedap en grup dinàmic de truges sobre sòl amb jaç de palla. Una decisió no massa extesa en granges del sud d’Europa on se sol preferir l’slat de formigó i/o superfície pavimentada, però imprescindible perquè aquesta explotació pogués aconseguir una millor gestió de les dejeccions ramaderes i assegurar una millora agronòmica de les terres de conreu. El treball conclou amb el seguiment del desenvolupament de l’alternativa escollida i una valoració dels canvis derivats de l’adaptació al Benestar Animal després del primer any i mig en ús. Les millores obtingudes en termes de maneig, funcionament de la granja, salut dels animals i índex productius són remarcables. D’una banda, cal destacar el fet de tenir un nombre de coixeres molt menor respecte les granges amb superfície dura, així com la facilitat i rapidesa en els parts degut al benestar. De l’altra,la reducció del volum de purí, l’increment de la fracció sòlida i la disponibilitat de compost per al camp. Per això, no és exagerat observar el jaç amb palla com una de les solucions més adequades i equilibrades per aquesta explotació que combina boví, porcí i terra per a la producció de cultius.

Effect of supplementation with MgCO3 and L-tryptophan on the welfare and on the carcass and meat quality of two halothane pig genotypes (NN and nn)

Sixty-one animals with different Halothane genes (homozygous halothane positive, n=34; and homozygous halothane negative, n=27) were fed with three diets (controlgroup, with no supplement; magnesium (Mg) group with 1.28g MgCO3/kg and tryptophan (Trp) group with 5g L-Trp/kg) during the last 5 days before slaughter. Animals were submitted to minimal stress ante mortem conditions. Pig behaviour was recorded at the experimental farm, raceway to the CO2 stunning system and during the stunning period. Corneal reflexes were recorded after stunning as well. There were no differences in feed intake among diets (p>0.05) during the 5 days of treatment. Thehalothane positive (nn) group had lower intake than the halothane negative (NN) group(p<0.01). The behaviour of the pigs in the raceway did not differ (p>0.05) among treatments or halothane genotype. A significant (p<0.001) interaction diet*halothane was found in the time to appear the first retreat attempt during the exposure to the CO2 system. In the nn group, the time of performing the first retreat attempt was later in the Mg (p<0.05) than the Control group. Moreover, in the Mg group, the nn had a later (p<0.05) first retreat attempt than the NN. Thus, Mg supplementation could have a positive effect on welfare of nn pigs. The nn had a lower proportion of animals thatshowed corneal reflexes after stunning than NN, indicating a higher effectiveness of thestunning method in nn pigs. Neither Mg nor Trp affected carcass quality and meat quality parameters, although significant differences were found between genotypes

Efecto de la suplementación con ácidos grasos de cadena media sobre la evolución de peso en lechones con poco peso al nacimiento

La evolución de peso en los períodos de lactación y transición de 583 lechones fue estudiada mediante un análisis estadístico, evaluando el efecto de la suplementación con ácidos grasos de cadena media (AGCM) en lechones con poco peso al nacimiento. 188 de los 375 lechones que nacieron con un peso al nacimiento (PN) <1250 g recibieron 3mL de AGCM cada 24 h durante los primeros 3 días de vida; su peso medio al destete (día 28) fue inferior respecto al grupo control (lechones no suplementados) (-114,17 g). No obstante, 106 de los 180 lechones nacidos con un PN <1000 g fueron suplementados, y su peso medio al destete y a finales de transición (día 63) fue superior respecto al grupo control (destete: +315,16 g; día 63: +775,47 g). Finalmente, los lechones suplementados con PN<800 g tuvieron los peores resultados: su diferencia de peso medio al destete fue de -177,58 g respecto al grupo control. Por lo tanto, en esta prueba fueron estudiados los lechones con un PN entre 800 y 999 g porque el grupo suplementado al destete tuvo una diferencia de peso medio considerable respecto al grupo control:+511,58 g. Asimismo, considerando una probabilidad de error inferior a 0,05, no hubieron diferencias significativas en las diferentes categorías de PN analizadas. De todas maneras, es importante destacar el alto grado de significación en la suplementación con AGCM en lechones con PN entre 800 y 999g (P=0,059). Por otra parte, el PN del grupo suplementado con PN<1000 g fue inferior que el del grupo no suplementado con PN<1000 g; esta diferencia de PN fue significativa (P=0,004) y como consecuencia el grado de significación en la suplementación con AGCM en lechones con PN entre 800 y 999 g fue inferior al esperado. Además, en esta prueba se incluyeron algunos resultados generales y también un análisis simple de supervivencia, aunque no era el objetivo principal

Biotecnología de la Reproducción Porcina: Estado actual y futuro de las técnicas de análisis seminal

Estat actual i futur de les tècniques d'anàlisi seminal